Updated on 2024/11/13

写真a

 
KAMIMURA Shinji
 
Organization
Faculty of Science and Engineering Professor
Other responsible organization
Biological Sciences Course of Graduate School of Science and Engineering, Master's Program
Biological Sciences Course of Graduate School of Science and Engineering, Doctoral Program
Contact information
The inquiry by e-mail is 《here
Profile
Interested in the molecular mechanisms of cell motility. In particular, in the structural background of motor proteins and cytoskeleton, and their evolutionary meaning. i.e., how the polypeptide components and their molecular configurations are optimized for their specific functions. Every molecular domain and its configuration dynamics should have its own meaning, which I would like to understand through a series of original experiments in the field of biophysics and cell physiology.
External link

Degree

  • 理学博士 ( 東京大学 )

  • 理学修士 ( 東京大学 )

Education

  • 1983.3
     

    The University of Tokyo   Graduate School, Division of Science   Department of Biological Sciences   doctor course   completed

  • 1980.3
     

    The University of Tokyo   Graduate School, Division of Science   master course   completed

  • 1978.3
     

    The University of Tokyo   Faculty of Science   Department of Biology   graduated

Research History

  • 2012.7 - 2012.9

    鳥取大学大学院 工学研究科 機械宇宙工学専攻 非常勤講師

  • 2012.7 - 2012.9

    鳥取大学大学院 工学研究科 機械宇宙工学専攻 非常勤講師

  • 2005.4 - 2012.3

    お茶の水女子大学 大学院理学系研究科 非常勤講師

  • 2005.4 - 2012.3

    お茶の水女子大学 大学院理学系研究科 非常勤講師

  • 2011.4 - 2011.9

    東京大学理学部 非常勤講師   Faculty of Science

  • 2006.4 - 2011.3

    日本女子大学 理学部 非常勤講師   Faculty of Science

  • 2008.4 - 2009.3

    名古屋大学 工学系研究科 非常勤講師

  • 2008.4 - 2009.3

    名古屋大学 工学系研究科 非常勤講師

  • 2008.4 -  

    Professor at the Department of Biological Sciences Chuo University

  • 2008.4 -  

    ~ 中央大学 理工学部 教授

  • 2008.4 -  

    - Professor at the Department of Biological Sciences Chuo University

  • 2007.4 - 2008.3

    "Associated professor at the Department of Biology, College of Arts and Sciences, University of Tokyo"   Graduate School of Arts and Sciences

  • 2007.4 - 2008.3

    Associated professor at the Department of Biology, College of Arts and Sciences, University of Tokyo

  • 1998.4 - 2007.3

    東京大学 大学院総合文化研究科 助教授

  • 2003.10 - 2004.3

    筑波大学生物科学系 非常勤講師   Institute of Biological Sciences

  • 2003.4 - 2003.9

    名古屋大学工学系研究科 非常勤講師

  • 2003.4 - 2003.9

    名古屋大学工学系研究科 非常勤講師

  • 1992.4 - 1998.3

    "Associated professor at the Department of Biology, College of Arts and Sciences, University of Tokyo"

  • 1992.4 - 1998.3

    Associated professor at the Department of Biology, College of Arts and Sciences, University of Tokyo

  • 1990.9 - 1992.3

    HFSP Long-Term Fellowships (LTF)

  • 1990.9 - 1992.3

    . HFSP Long-Term Fellowships (LTF)

  • 1985.4 - 1992.3

    "Assistant at the Department of Biology, College of Arts and Sciences, University of Tokyo"

  • 1985.4 - 1992.3

    Assistant at the Department of Biology, College of Arts and Sciences, University of Tokyo

  • 1985.4 - 1990.3

    放送大学 東京学習センター 非常勤講師

  • 1983.4 - 1985.3

    "Research fellow at ERATO-project, Bioholonics Project of Research Development Corporation of Japan"

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Professional Memberships

  • Zoological Society of Japan

  • 日本生物物理学会

  • Biophysical Society (USA)

  • エアロアクアバイオメカニズム学会

  • Zoological Society of Japan

  • Biophysical Society (USA)

  • Society of Aero Aqua Biomechanism

  • Zoological Society of Japan

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Research Interests

  • cilia

  • flagella

  • "flagella, cilia, microtubules, cell motility, cytoskeleton, dynein, signal transduction, primary cilia"

  • 構造生物学

  • Cell physiology

  • Structural Biology

  • ダイニン

  • 細胞骨格

  • 鞭毛

  • 微小管

  • 細胞運動

  • 繊毛

  • プライマリ繊毛

  • Cell physiology

  • シグナル伝達

  • signal transduction

  • primary cilia

  • cytoskeleton

  • dynein

  • microtubules

  • cell motility

Research Areas

  • Life Science / Cell biology  / Cell biology

  • Life Science / Cell biology  / Cell physiology

  • Life Science / Structural biochemistry  / Structural biology

  • Nanotechnology/Materials / Nanobioscience

  • Life Science / Biophysics  / Biophysics

  • Nanotechnology/Materials / Nanomaterials

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Papers

  • Structural insight into the stabilization of microtubules by taxanes

    Andrea E Prota, Daniel Lucena-Agell, Yuntao Ma, Juan Estevez-Gallego, Shuo Li, Katja Bargsten, Fernando Josa-Prado, Karl-Heinz Altmann, Natacha Gaillard, Shinji Kamimura, Tobias Mühlethaler, Federico Gago, Maria A Oliva, Michel O Steinmetz, Wei-Shuo Fang, J Fernando Díaz

    eLIFE   2023.3

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    DOI: 10.7554/eLife.84791

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  • Chemical modulation of microtubule structure through the laulimalide/peloruside site. Reviewed International journal

    Juan Estévez-Gallego, Beatriz Álvarez-Bernad, Benet Pera, Christoph Wullschleger, Olivier Raes, Dirk Menche, Juan Carlos Martínez, Daniel Lucena-Agell, Andrea E Prota, Francesca Bonato, Katja Bargsten, Jelle Cornelus, Juan Francisco Giménez-Abián, Peter Northcote, Michel O Steinmetz, Shinji Kamimura, Karl-Heinz Altmann, Ian Paterson, Federico Gago, Johan Van der Eycken, J Fernando Díaz, María Ángela Oliva

    Structure (London, England : 1993)   31 ( 1 )   88 - 99   2023.1

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    Taxanes are microtubule-stabilizing agents used in the treatment of many solid tumors, but they often involve side effects affecting the peripheral nervous system. It has been proposed that this could be related to structural modifications on the filament upon drug binding. Alternatively, laulimalide and peloruside bind to a different site also inducing stabilization, but they have not been exploited in clinics. Here, we use a combination of the parental natural compounds and derived analogs to unravel the stabilization mechanism through this site. These drugs settle lateral interactions without engaging the M loop, which is part of the key and lock involved in the inter-protofilament contacts. Importantly, these drugs can modulate the angle between protofilaments, producing microtubules of different diameters. Among the compounds studied, we have found some showing low cytotoxicity and able to induce stabilization without compromising microtubule native structure. This opens the window of new applications for microtubule-stabilizing agents beyond cancer treatment.

    DOI: 10.1016/j.str.2022.11.006

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  • Alternative Approaches to Understand Microtubule Cap Morphology and Function Invited

    Oliva, M.A, Gago, F, Kamimura, S, Díaz, J.F

    ACS Omega   2023.1

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    DOI: 10.1021/acsomega.2c06926

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  • Metabolomic profiling upon external digestion in larvae of diving beetles: Cybister Curtis, 1827, Dytiscus Linnaeus, 1758, and Hydaticus Leach, 1817 (Coleoptera: Dytiscidae) Reviewed

    Toshio Inoda, Shinji Kamimura

    Aquatic Insects   2022.9

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    The SDS.PAGE analysis of prey animals (fly larvae) during external digestion by aquatic predators, the larvae of selected species in dytiscid genera, showed a major component of protein with a molecular weight of ca. 20 kDa. In addition, the analysis of third instar larvae of Cybister brevis Aube, 1838, by nanoLC.ESI.Q-TOF/MS/MS revealed that the digested body fluid includes a polypeptide with a sequence of 97 amino acids corresponding to hemocyanin N (51% matched by Mascot and BLAST searches) and hemocyanin M (12% matched) derived from flies. We also found evidence indicating that beetle larvae repeatedly release digestive enzymes at least twice while consuming the fly bodies. These results suggest that the digested polypeptides were derived from ubiquitous high molecular substances such as arylphorin subunit C223 precursor included in the body fluid of the fly (Calliphora vicina Robineau-Desvoidy, 1830) that were produced during external digestion by diving beetle larvae.

    DOI: 10.1080/01650424.2022.2076883

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  • GTP-dependent formation of straight tubulin oligomers leads to microtubule nucleation Reviewed International journal

    Rie Ayukawa, Seigo Iwata, Hiroshi Imai, Shinji Kamimura, Masahito Hayashi, Kien Xuan Ngo, Itsushi Minoura, Seiichi Uchimura, Tsukasa Makino, Mikako Shirouzu, Hideki Shigematsu, Ken Sekimoto, Benoît Gigant, Etsuko Muto

    Journal of Cell Biology   220 ( 4 )   e202007033   2021.4

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ROCKEFELLER UNIV PRESS  

    Nucleation of microtubules (MTs) is essential for cellular activities, but its mechanism is unknown because of the difficulty involved in capturing rare stochastic events in the early stage of polymerization. Here, combining rapid flush negative stain electron microscopy (EM) and kinetic analysis, we demonstrate that the formation of straight oligomers of critical size is essential for nucleation. Both GDP and GTP tubulin form single-stranded oligomers with a broad range of curvatures, but upon nucleation, the curvature distribution of GTP oligomers is shifted to produce a minor population of straight oligomers. With tubulin having the Y222F mutation in the β subunit, the proportion of straight oligomers increases and nucleation accelerates. Our results support a model in which GTP binding generates a minor population of straight oligomers compatible with lateral association and further growth to MTs. This study suggests that cellular factors involved in nucleation promote it via stabilization of straight oligomers.

    DOI: 10.1083/jcb.202007033

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  • GTP-Dependent Formation of Straight Oligomers Leads to Nucleation of Microtubules

    Etsuko Muto, Rie Ayukawa, Seigo Iwata, Hiroshi Imai, Shinji Kamimura, Ken Sekimoto, Gigant Benoit

    BIOPHYSICAL JOURNAL   120 ( 3 )   12A - 12A   2021.2

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  • SAXS Signals from Double-Stranded DNA under Shear-Flow Conditions Reviewed

    Yosuke Fujita, Takako Kato-Minoura, Longo Alessandro, Yuko Wada, Toshiki Yagi, Hiroyuki Iwamoto, Ritsu Kamiya, Shinji Kamimur

    SPring-8 Section A: Scientific Research Report   8 ( 3 )   2020.10

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    SAXS signals from the DNA of salmon spermatozoa in an aqueous solution were analyzed using a rheometer-device, which controlled the rate of shearing during SAXS observations. When shearing rate > 1000 s−1 was applied to the DNA suspension, DNA strands were not aligned as observed with other biological filaments, but a stable diffraction peak approximately ranging 1.7–1.8 [nm−1] of Q-values were observed. We compared our observation with simulated SAXS signals.

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  • Development of phase-contrast/dark-field microscopy with scanning laser illumination Reviewed

    Tsuyoshi SATO, Tomotaka SHIRANE, Yuuko WADA, Hiroshi IMAI, Shinji KAMIMURA

    Journal of the Institute of Science and Engineering. Chuo University   2019 ( 25 )   93 - 103   2020.3

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Chuo Uiversity  

    In this study, we designed a novel optics to scan laser light on the front focal plane of condenser lens, i.e., aperture plane, that is, with light sources placed optically at an optically infinite distance from specimens. Diatom standard specimens were used for the observation in both the phase contrast image and the dark field microscopy, and we could reduce speckle noise under applicable magnitude. Although we could not solve the problem of biased brightness of illumination depending on specimen angles, the new optics presented here is expected to be useful for the observation of fine structures with high image brightness.

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  • Structural model for differential cap maturation at growing microtubule ends Reviewed International journal

    Juan Estevez-Gallego, Fernando Josa-Prado, Siou Ku, Ruben M. Buey, Francisco A. Balaguer, Andrea E. Prota, Daniel Lucena-Agell, Christina Kamma-Lorger, Toshiki Yagi, Hiroyuki Iwamoto, Laurence Duchesne, Isabel Barasoain, Michel O. Steinmetz, Denis Chretien, Shinji Kamimura, J. Fernando Diaz, Maria A. Oliva

    ELIFE   9   e50155   2020.3

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELIFE SCIENCES PUBLICATIONS LTD  

    Microtubules (MTs) are hollow cylinders made of tubulin, a GTPase responsible for essential functions during cell growth and division, and thus, key target for anti-tumor drugs. In MTs, GTP hydrolysis triggers structural changes in the lattice, which are responsible for interaction with regulatory factors. The stabilizing GTP-cap is a hallmark of MTs and the mechanism of the chemical-structural link between the GTP hydrolysis site and the MT lattice is a matter of debate. We have analyzed the structure of tubulin and MTs assembled in the presence of fluoride salts that mimic the GTP-bound and GDP•Pi transition states. Our results challenge current models because tubulin does not change axial length upon GTP hydrolysis. Moreover, analysis of the structure of MTs assembled in the presence of several nucleotide analogues and of taxol allows us to propose that previously described lattice expansion could be a post-hydrolysis stage involved in Pi release.

    DOI: 10.7554/eLife.50155

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  • 分子モーター研究の新しい方向 キネシンによる微小管の構造変化

    島 知弘, 森川 真夏, 金城 純一, 神原 丈敏, 上村 慎治, 八木 俊樹, 岩本 裕之, 市村 垂生, 渡邉 朋信, 上村 想太郎, 仁田 亮, 岡田 康志, 廣川 信隆

    日本細胞生物学会大会講演要旨集   69回   39 - 39   2017.5

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  • 1P143 Paclitaxel induces the quick elongation of tubulin dimer periodicity in microtubules(11. Molecular motor,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Kamimura Shinji, Kiyohara Megumi, Nakazawa Nobumasa, Fujita Yosuke, Wada Yuuko, Yagi Toshiki, Iwamoto Hiroyuki

    Seibutsu Butsuri   54 ( 1 )   S164   2014

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    DOI: 10.2142/biophys.54.S164_5

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  • 1P197 Flagellar central structures regulate the dynein motor activity through the change of axonemal diameter(12.Cell biology,Poster,The 51st Annual Meeting of the Biophysical Society of Japan)

    Yagi Toshiki, Fujita Yosuke, Kamimura Shinji, Iwamoto Hiroyuki

    Seibutsu Butsuri   53 ( 1 )   S138   2013

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    DOI: 10.2142/biophys.53.S138_4

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  • 1P213 A new function of cilia,cell-signaling enhancer, revealed by the simulation analysis of intra-ciliary diffusion(Cell biology,The 48th Annual Meeting of the Biophysical Society of Japan)

    Takao Daisuke, Kamimura Shinji

    Seibutsu Butsuri   50 ( 2 )   S56 - S57   2010

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    DOI: 10.2142/biophys.50.S56_6

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  • FRAP Analysis Combined With A Single-cell Electroporation Technique In Sea-urchin Spermatozoa

    Daisuke Takao, Shinji Kamimura

    BIOPHYSICAL JOURNAL   96 ( 3 )   520A - 520A   2009.2

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    Language:English   Publisher:CELL PRESS  

    DOI: 10.1016/j.bpj.2008.12.2680

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  • 3TA4-02 X-ray diffraction analysis of the dynamic features of microtubule structure(The 47th Annual Meeting of the Biophysical Society of Japan)

    Kamimura Shinji, Iwamoto Hiroyuki, Miyashiro Daisuke

    Seibutsu Butsuri   49   S56   2009

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    DOI: 10.2142/biophys.49.S56_4

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  • Erratum: Temperature-dependent regulation of reproduction in the diving beetle Dytiscus sharpi (Coieoptera: Dytiscidae)(Zoological Science(November, 2007), 24, (11), (1115-1121))

    Inoda, T., Tajima, F., Taniguchi, H., Saeki, M., Numakura, K., Hasegawa, M., Kamimura, S.

    Zoological Science   25 ( 5 )   560   2008.5

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    DOI: 10.2108/zsj.25.560

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  • A new microscope optics for laser dark-field illumination applied to high precision two dimensional measurement of specimen displacement Reviewed International journal

    Noda, N., Kamimura, S.

    Review of Scientific Instruments   79 ( 2 )   023704 - 023704   2008.2

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER INST PHYSICS  

    With conventional light microscopy, precision in the measurement of the displacement of a specimen depends on the signal-to-noise ratio when we measure the light intensity of magnified images. This implies that, for the improvement of precision, getting brighter images and reducing background light noise are both inevitably required. For this purpose, we developed a new optics for laser dark-field illumination. For the microscopy, we used a laser beam and a pair of axicons (conical lenses) to get an optimal condition for dark-field observations. The optics was applied to measuring two dimensional microbead displacements with subnanometer precision. The bandwidth of our detection system overall was 10 kHz. Over most of this bandwidth, the observed noise level was as small as 0.1 nm/radicalHz.

    DOI: 10.1063/1.2839914

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  • Preface

    Naomi Kato, Shinji Kamimura

    Bio-Mechanisms of Swimming and Flying: Fluid Dynamics, Biomimetic Robots, and Sports Science   2008

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    Publishing type:Research paper (international conference proceedings)  

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  • 2P-206 Chemoattractant sensing mechanism in sperm : Experimental and simulation study(The 46th Annual Meeting of the Biophysical Society of Japan)

    Shiba Kogiku, Miyashiro Daisuke, Kamimura Shinji, Yoshida Manabu

    Seibutsu Butsuri   48   S106 - S107   2008

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    DOI: 10.2142/biophys.48.S106_6

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  • 2P-342 Nucleotide caps and the stability of cytoskeletal fibrous proteins : A high-pressure small-angle x-ray scattering study(The 46th Annual Meeting of the Biophysical Society of Japan)

    Fujisawa Tetsuro, Matsuo Hiroshi, Iwasa Mitsusada, Erickson Harold P., Kamimura Shinji, Maeda Yuichiro, Popp David

    Seibutsu Butsuri   48   S180   2008

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    DOI: 10.2142/biophys.48.S180_3

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  • 2P166 Helical arrangement of axonemal components in sea-urchin sperm flagella revealed(Molecular motors,Oral Presentations)

    Kamimura Shinji, Iwamoto Hiroyuki

    Seibutsu Butsuri   47   S154   2007

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    DOI: 10.2142/biophys.47.S154_3

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  • A novel flow-alignment technique for the X-ray diffraction structure analysis of sea-urchin sperm flagella

    Shinji Kamimura, Takaaki Sugiyama, Yasunobu Sugimoto, Katsuzo Wakabayashi

    ZOOLOGICAL SCIENCE   23 ( 12 )   1167 - 1167   2006.12

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  • 2P259 X-ray diffraction from dynein motors and microtubules in seaurchin sperm flagellar axonemes(39. Cell motility,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

    Kamimura Shinji, Iwamoto Hiroyuki, Fujisawa Tetsuro

    Seibutsu Butsuri   46 ( 2 )   S360   2006

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    DOI: 10.2142/biophys.46.S360_3

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  • Analysis of the flagellar motion of Raphidophyceae, Chattonella antiqua

    Shinji Kamimura, Naomi Katoh, Tatsuro Akiba

    ZOOLOGICAL SCIENCE   22 ( 12 )   1446 - 1446   2005.12

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  • Temperature regulates the summer reproductive diapause in diving beetles, Dytiscus sharpi (Coleoptera : Dytiscidae)

    Toshio Inoda, Fumitada Tajima, Hiroshi Taniguchi, Motoyuki Saeki, Kazuki Numakura, Shinji Kamimura

    ZOOLOGICAL SCIENCE   21 ( 12 )   1326 - 1326   2004.12

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  • A new optical system for the determination of pH change in a sea-urchin sperm cell

    Daisuke Takao, Shinji Kamimura

    ZOOLOGICAL SCIENCE   21 ( 12 )   1283 - 1283   2004.12

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  • 3P200 X-ray diffraction analysis of the dynamic structural change of sea-urchin sperm flagellar axonemes

    Kamimura S., Wakamatsu J., Tamura T., Fujisawa T., Iwamoto H.

    Seibutsu Butsuri   44   S239   2004

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    DOI: 10.2142/biophys.44.S239_4

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  • Dynein-ADP as a force-generating intermediate revealed by a rapid reactivation of flagellar axoneme

    Tomomi Tani, Shinji Kamimura

    Biophysical Journal   77 ( 3 )   1518 - 1527   1999.9

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    Fragmented flagellar axonemes of sand dollar spermatozoa were reactivated by rapid photolysis of caged ATP. After a time lag of 10 ms, axonemes treated with protease started sliding disintegration. Axonemes without protease digestion started nanometer-scale high-frequency oscillation after a similar time lag. Force development in the sliding disintegration was measured with a flexible glass needle and its time course was corresponded well to that of the dynein-ADP intermediate production estimated using kinetic rates previously reported. However, with a high concentration (~80 μM) of vanadate, which binds to the dynein-ADP intermediate and forms a stable complex of dynein-ADP-vanadate, the time course of force development in sliding disintegration was not affected at all. In the case of high frequency oscillation, the time lag to start the oscillation, the initial amplitude, and the initial frequency were not affected by vanadate, though the oscillation once started was damped more quickly at higher concentrations of vanadate. These results suggest that during the initial turnover of ATP hydrolysis, force generation of dynein is not blocked by vanadate. A vanadate-insensitive dynein-ADP is postulated as a force-generating intermediate.

    DOI: 10.1016/S0006-3495(99)76999-7

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  • Structural fluctuation and heterogeneity of tubulin arrangement in microtubules Invited

    Shinji Kamimura

    THE CELL   55 ( 3 )   54 - 57   1900.3

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    Authorship:Lead author   Language:Japanese  

    File: THECELL_SKamimura2023Mar.pdf

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Books

  • 高等学校理科用 文部科学省検定済教科書 生物 東京書籍(生物 301)

    浅島誠(第3編)

    東京書籍  2024.2  ( ISBN:4487187494

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    Total pages:479   Language:Japanese  

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  • 分子細胞生物学

    Lodish, Harvey F., Berk, Arnold, Kaiser, Chris, Krieger, Monty, Bretscher, Anthony, Ploegh, Hidde L., Martin, Kelsey C., Yaffe, Michael B., Amon, Angelika, 岩井, 佳子, 上村, 慎治, 北川, 大樹, 齋藤, 康太, 坪井, 貴司, 富田, 泰輔, 名黒, 功, 仁科, 博史, 宮澤, 恵二, 山本, 啓一, 若林, 憲一, 堅田, 利明, 須藤, 和夫( Role: Translator/Editor10.生体膜の構造, 17.細胞の構築と運動 I:ミクロフィラメント, 20.細胞から組織への集成)

    東京化学同人  2023.7  ( ISBN:9784807920518

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    Total pages:xxxv, 1074p   Language:Japanese  

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  • 動物の事典

    上村慎治( Role: Joint editor章担当編集者)

    朝倉書店  2020.11 

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    Total pages:746   Responsible for pages:3ページ   Language:Japanese   Book type:Dictionary, encyclopedia

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  • 動物学の百科事典

    Kamimura S( Role: Edit第5章編集、導入、「細胞骨格」担当)

    Maruzen Pub Co  2018.9 

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    Total pages:770   Responsible for pages:3   Language:Japanese  

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  • 動物学の百科事典

    丸善出版株式会社  2018.9 

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  • 改訂 生物 [2 東書 生物 306] 高校教科書 文部科学省検定済教科書

    浅島, 誠(第3編)

    東京書籍  2018.2  ( ISBN:4487165555

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    Total pages:3, 480, 4, 2p   Language:Japanese  

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  • 改訂 生物基礎 [2東書/生基311] 文部科学省検定済教科書

    浅島, 誠(第3編)

    東京書籍  2017.2  ( ISBN:4487165490

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    Total pages:3, 248, 2p   Language:Japanese  

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  • Cain Biology

    S. Kamimura( Role: Supervisor (editorial)動物の反応と調節)

    Tokyo Kagaku Dojin  2014.9 

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    Total pages:728   Responsible for pages:60   Language:Japanese   Book type:Scholarly book

    http://www.tkd-pbl.com/book/b182504.html

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  • Cain Biology

    Tokyo Kagaku Dojin  2014.9 

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  • Cain Biology

    Shinji Kamimura, Tomko Noguchi, Mariko Kamimura( Role: Supervisor (editorial)動物の反応と調節)

    Tokyo Kagaku Dojin  2012.2 

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    Total pages:360   Responsible for pages:360   Language:Japanese   Book type:Scholarly book

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  • Cain Biology

    Tokyo Kagaku Dojin  2012.2 

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  • 生き物に学ぶ泳ぎと飛行のしくみ(第4章 微生物の運動)

    エアロ・アクアバイオメカニズム研究会(編)  2010.8 

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  • 生き物に学ぶ泳ぎと飛行のしくみ(第4章 微生物の運動)

    ( Role: Sole author)

    エアロ・アクアバイオメカニズム研究会(編)  2010.8 

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    Language:Japanese   Book type:Scholarly book

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  • 生物学辞典

    石川統, 黒岩常祥, 塩見正衛, 松本忠夫, 守隆夫, 八杉貞雄, 山本正幸( Role: Joint editor)

    東京化学同人  2009.10 

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    Total pages:1620   Responsible for pages:40   Language:Japanese   Book type:Scholarly book

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  • Bio-mechanisms of swimming and flying: Fluid dynamics, biomimetic robots, and sports science.

    Kato, N, Kamimura,S( Role: Sole author)

    Springer  2007.4 

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    Total pages:403   Language:English   Book type:Scholarly book

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  • Bio-mechanisms of swimming and flying: Fluid dynamics, biomimetic robots, and sports science.

    Springer  2007.4 

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  • Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport. International journal

    Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Sotaro Uemura, Hideki Shigematsu, Mikako Shirouzu, Taro Ichimura, Tomonobu M Watanabe, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa

    The Journal of cell biology   217 ( 12 )   4164 - 4183   2018.12

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    Kinesin-1, the founding member of the kinesin superfamily of proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here, we report that kinesin-1 binding changes the microtubule lattice and promotes further kinesin-1 binding. This high-affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low-affinity state by washing out the binding kinesin-1 or by the binding of non-hydrolyzable ATP analogue AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy, and second-harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin-1 to GDP microtubules changes the conformation of the GDP microtubule to a conformation resembling the GTP microtubule.

    DOI: 10.1083/jcb.201711178

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  • Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport International journal

    Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Sotaro Uemura, Hideki Shigematsu, Mikako Shirouzu, Taro Ichimura, Tomonobu M. Watanabe, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa

    JOURNAL OF CELL BIOLOGY   217 ( 12 )   4164 - 4183   2018.12

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    Language:English   Publisher:ROCKEFELLER UNIV PRESS  

    Kinesin-1, the founding member of the kinesin superfamily of proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here, we report that kinesin-1 binding changes the microtubule lattice and promotes further kinesin-1 binding. This high-affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low-affinity state by washing out the binding kinesin-1 or by the binding of non-hydrolyzable ATP analogue AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy, and second-harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin-1 to GDP microtubules changes the conformation of the GDP microtubule to a conformation resembling the GTP microtubule.

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  • Simulation of intra-ciliary diffusion suggests a novel role of primary cilia as a cell-signaling enhancer. Reviewed

    Daisuke Takao, Shinji Kamimura

    Development, growth & differentiation   59 ( 5 )   415 - 422   2017.6

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    Besides the role to generate a fluid flow in the surrounding medium, eukaryotic cilia have a crucial function in sensing external signals such as chemical or mechanical stimuli. A large body of work has shown that cilia are frequently found in various types of sensory cells and are closely related to many regulatory mechanisms in differentiation and development. However, we do not yet have a definitive answer to the fundamental question, 'why cilia?.' It has been a long-standing mystery why cells use cilia for sensing external signals. To shed light on this, we sought to describe the kinetics of signaling with theoretical approaches. Based on the results, here we propose a new role of cilia as a cell-signaling enhancer. The enhancing effect comes from restricted volume for the free intra-ciliary diffusion of molecules due to the cylindrical shape of cilia, which can facilitate quick accumulation of intracellular signaling molecules. Our simulations demonstrate that both the rate and amplitude of response in signal transduction depend on where the membrane receptors or channels are located along the ciliary shaft. In addition, the calculated transfer function of cilia regarded as a

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  • Simulation of intra-ciliary diffusion suggests a novel role of primary cilia as a cell-signaling enhancer

    Daisuke Takao, Shinji Kamimura

    DEVELOPMENT GROWTH & DIFFERENTIATION   59 ( 5 )   415 - 422   2017.6

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    Besides the role to generate a fluid flow in the surrounding medium, eukaryotic cilia have a crucial function in sensing external signals such as chemical or mechanical stimuli. A large body of work has shown that cilia are frequently found in various types of sensory cells and are closely related to many regulatory mechanisms in differentiation and development. However, we do not yet have a definitive answer to the fundamental question, "why cilia?" It has been a long-standing mystery why cells use cilia for sensing external signals. To shed light on this, we sought to describe the kinetics of signaling with theoretical approaches. Based on the results, here we propose a new role of cilia as a cell-signaling enhancer. The enhancing effect comes from restricted volume for the free intra-ciliary diffusion of molecules due to the cylindrical shape of cilia, which can facilitate quick accumulation of intracellular signaling molecules. Our simulations demonstrate that both the rate and amplitude of response in signal transduction depend on where the membrane receptors or channels are located along the ciliary shaft. In addition, the calculated transfer function of cilia regarded as a transmitter of external signals also suggests the properties of cilia as a signal enhancer. Since such unique composition of receptors and channels in cilia is found in various types of eukaryotic cells, signal enhancing is presumably one of the most essential and conserved roles of cilia.

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  • X-ray fiber diffraction: a tool for understanding the structural dynamics of tubulin dimers in native microtubules.

    Shinji Kamimura

    SPring-8/SACLA Research Frontiers 2016   2016   32 - 33   2017.6

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    Language:English   Publishing type:Article, review, commentary, editorial, etc. (other)   Publisher:JASRI  

    When we are going to know structural details of biological molecules, there would be two possibilities; electron microscopy and X-ray crystallography. Owing to remarkable improvements in cryo-electron microscopy techniques using direct electron detectors, impressive progresses showing high-resolution (0.3 nm) images of various biological molecules in action are now appearing regularly in leading journals. Since details of protein architectures at the atomic scale have been obtained by X-ray crystallography, we can now combine them with electron micrographs to gain more direct insights into the functions of biomolecules than we expected by conventional tools used over a decade ago. This would also be the case for X-ray fiber diffraction/X-ray solution scattering analysis, which we expect will become the third major method used for structural biology.

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  • X-ray fiber diffraction: a tool for understanding the structural dynamics of tubulin dimers in native microtubules.

    Shinji Kamimura

    SPring-8/SACLA Research Frontiers 2016   32 - 33   2017.6

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  • X-ray fiber diffraction analysis shows dynamic changes in axial tubulin repeats in native microtubules depending on paclitaxel content, temperature and GTP-hydrolysis. International journal

    Shinji Kamimura, Yosuke Fujita, Yuuko Wada, Toshiki Yagi, Hiroyuki Iwamoto

    Cytoskeleton (Hoboken, N.J.)   73 ( 3 )   131 - 44   2016.3

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    Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most fundamental questions is the association of microtubule dynamics with the molecular conformation of tubulin within microtubules. To address this issue, we applied a new technique for the rapid shear-flow alignment of biological filaments, enabling us to acquire the structural periodicity data of microtubules by X-ray fiber diffraction under various physiological conditions. We classified microtubules into three main groups on the basis of distinct axial tubulin periodicities and mean microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel, a microtubule stabilizer. Paclitaxel induced rapid changes in tubulin axial repeats in a cooperative manner. This is the first demonstration of dynamic changes of axial tubulin repeats within native microtubules without fixation. We also found extraordinary features of negative thermal expansion of axial tubulin repeats in both paclitaxel-stabilized and GMPCPP-containing microtubules. Our results suggest that even in assembled microtubules, both GTP- and GDP-tubulin dimers can undergo dynamic conversion between at least two different states: short and long configurations. (c) 2016 Wiley Periodicals, Inc.

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  • X-ray fiber diffraction analysis shows dynamic changes in axial tubulin repeats in native microtubules depending on paclitaxel content, temperature and GTP-hydrolysis

    Shinji Kamimura

    Cytoskeleton   73 ( 3 )   131 - 144   2016.3

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    Language:English   Publisher:WILEY  

    Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most fundamental questions is the association of microtubule dynamics with the molecular conformation of tubulin within microtubules. To address this issue, we applied a new technique for the rapid shear-flow alignment of biological filaments, enabling us to acquire the structural periodicity data of microtubules by X-ray fiber diffraction under various physiological conditions. We classified microtubules into three main groups on the basis of distinct axial tubulin periodicities and mean microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel, a microtubule stabilizer. Paclitaxel induced rapid changes in tubulin axial repeats in a cooperative manner. This is the first demonstration of dynamic changes of axial tubulin repeats within native microtubules without fixation. We also found extraordinary features of negative thermal expansion of axial tubulin repeats in both paclitaxel-stabilized and GMPCPP-containing microtubules. Our results suggest that even in assembled microtubules, both GTP- and GDP-tubulin dimers can undergo dynamic conversion between at least two different states: short and long configurations. (c) 2016 Wiley Periodicals, Inc.

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  • A simple method for nanobubble generation and stability of the bubbles

    Ohmori, M, Haruta, K, Kamimura, S, Koike, H, Uchida, Takeyama, H

    Journal of Environmental Biotechnology   41 - 44   2015.10

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  • A simple method for nanobubble generation and stability of the bubbles Reviewed

    Ohmori, M, Haruta, K, Kamimura, S, Koike, H, Uchida, Takeyama, H

    Journal of Environmental Biotechnology   15 ( 1 )   41 - 44   2015.10

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    We could generate very small air-nanobubbles, having diameters smaller than 100 nm, using a household hand mixer. The bubbles were stable in water containing 1% ethanol more than one month and show zeta potential of -30 to -40 mV. The addition of NaCl to nonobubble-water caused changes in the diameter and the number of bubbles, larger in size and less in numbers.

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  • X-Ray Fiber Diffraction Recordings from Oriented Demembranated Chlamydomonas Flagellar Axonemes. International journal

    Shiori Toba, Hiroyuki Iwamoto, Shinji Kamimura, Kazuhiro Oiwa

    Biophysical journal   108 ( 12 )   2843 - 53   2015.6

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    The high homology of its axonemal components with humans and a large repertoire of axonemal mutants make Chlamydomonas a useful model system for experiments on the structure and function of eukaryotic cilia and flagella. Using this organism, we explored the spatial arrangement of axonemal components under physiological conditions by small-angle x-ray fiber diffraction. Axonemes were oriented in physiological solution by continuous shear flow and exposed to intense and stable x rays generated in the synchrotron radiation facility SPring-8, BL45XU. We compared diffraction patterns from axonemes isolated from wild-type and mutant strains lacking the whole outer arm (oda1), radial spoke (pf14), central apparatus (pf18), or the a-chain of the outer arm dynein (oda11). Diffraction of the axonemes showed a series of well-defined meridional/layer-line and equatorial reflections. Diffraction patterns from mutant axonemes exhibited a systematic loss/attenuation of meridional/layer-line reflections, making it possible to determine the origin of various reflections. The 1/24 and 1/12 nm(-1) meridional reflections of oda1 and oda11 were much weaker than those of the wild-type, suggesting that the outer dynein arms are the main contributor to these reflections. The weaker 1/32 and 1/13.7 nm(-1) meridional reflections from pf14 compared with the wild-type suggest that these reflections come mainly from the radial spokes. The limited contribution of the central pair apparatus to the diffraction patterns was confirmed by the similarity between the patterns of the wild-type and pf18. The equatorial reflections were complex, but a comparison with electron micrograph-based models allowed the density of each axonemal component to be estimated. Addition of ATP to rigor-state axonemes also resulted in subtle changes in equatorial intensity profiles, which could report nucleotide-dependent structural changes of the dynein arms. The first detailed description of axonemal reflections presented here serves as a landmark for further x-ray diffraction studies to monitor the action of constituent proteins in functional axonemes.

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  • X-Ray Fiber Diffraction Recordings from Oriented Demembranated Chlamydomonas Flagellar Axonemes

    Toba, S., Iwamoto, H., Kamimura, S., Oiwa, K.

    Biophysical Journal   108 ( 12 )   2843 - 2853   2015.6

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    The high homology of its axonemal components with humans and a large repertoire of axonemal mutants make Chlamydomonas a useful model system for experiments on the structure and function of eukaryotic cilia and flagella. Using this organism, we explored the spatial arrangement of axonemal components under physiological conditions by small-angle x-ray fiber diffraction. Axonemes were oriented in physiological solution by continuous shear flow and exposed to intense and stable x rays generated in the synchrotron radiation facility SPring-8, BL45XU. We compared diffraction patterns from axonemes isolated from wild-type and mutant strains lacking the whole outer arm (oda1), radial spoke (pf14), central apparatus (pf18), or the a-chain of the outer arm dynein (oda11). Diffraction of the axonemes showed a series of well-defined meridional/layer-line and equatorial reflections. Diffraction patterns from mutant axonemes exhibited a systematic loss/attenuation of meridional/layer-line reflections, making it possible to determine the origin of various reflections. The 1/24 and 1/12 nm(-1) meridional reflections of oda1 and oda11 were much weaker than those of the wild-type, suggesting that the outer dynein arms are the main contributor to these reflections. The weaker 1/32 and 1/13.7 nm(-1) meridional reflections from pf14 compared with the wild-type suggest that these reflections come mainly from the radial spokes. The limited contribution of the central pair apparatus to the diffraction patterns was confirmed by the similarity between the patterns of the wild-type and pf18. The equatorial reflections were complex, but a comparison with electron micrograph-based models allowed the density of each axonemal component to be estimated. Addition of ATP to rigor-state axonemes also resulted in subtle changes in equatorial intensity profiles, which could report nucleotide-dependent structural changes of the dynein arms. The first detailed description of axonemal reflections presented here serves as a landmark for further x-ray diffraction studies to monitor the action of constituent proteins in functional axonemes.

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  • Effects of the dynein inhibitor ciliobrevin on the flagellar motility of sea urchin spermatozoa

    Yuuko Wada, Shoji A. Baba, Shinji Kamimura

    CYTOSKELETON   72 ( 4 )   182 - 192   2015.4

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    Ciliobrevin has recently been found to be a membrane-permeable inhibitor that is specific to AAA+ molecular motors such as cytoplasmic dyneins. In this study, we investigated how ciliobrevin inhibited the motility of sperm from sea urchins: Hemicentrotus pulcherrimus, Pseudocentrotus depressus, and Anthocidaris crassispina. After application of 100 M of ciliobrevin A to live spermatozoa, swimming speed decreased gradually and flagellar motion stopped almost completely within 5 to 10 min. This inhibition was reversible and the frequency of flagellar beating was reduced in a concentration-dependent manner. Ciliobrevin had similar inhibitory effects on the flagellar beating of demembranated and reactivated sperm and the sliding disintegration of trypsin-treated axonemes. We also analyzed the curvature and shear angle of the beating flagella and found that the proximal region of the sperm flagellum was less sensitive to ciliobrevin compared with more distal regions, where bending motions were blocked completely. Interestingly, the shear angle analysis of flagellar motility showed that ciliobrevin induced highly asymmetric bends in the proximal region of the flagellum. These results suggest that there is heterogeneity in the inhibitory thresholds of dynein motors, which depend on the regions along the flagellar shaft (proximal or distal) and on the sites of doublets in the flagellar cross-section (doublet numbers). We expect that it will be possible to map the functional differences in dynein subtypes along and/or around the cross-sections of flagellar axonemes by analyzing the inhibitory effects of ciliobrevin. (c) 2015 Wiley Periodicals, Inc.

    DOI: 10.1002/cm.21218

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  • Effects of the dynein inhibitor ciliobrevin on the flagellar motility of sea urchin spermatozoa. International journal

    Yuuko Wada, Shoji A Baba, Shinji Kamimura

    Cytoskeleton (Hoboken, N.J.)   72 ( 4 )   182 - 92   2015.4

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    Ciliobrevin has recently been found to be a membrane-permeable inhibitor that is specific to AAA+ molecular motors such as cytoplasmic dyneins. In this study, we investigated how ciliobrevin inhibited the motility of sperm from sea urchins: Hemicentrotus pulcherrimus, Pseudocentrotus depressus, and Anthocidaris crassispina. After application of 100 M of ciliobrevin A to live spermatozoa, swimming speed decreased gradually and flagellar motion stopped almost completely within 5 to 10 min. This inhibition was reversible and the frequency of flagellar beating was reduced in a concentration-dependent manner. Ciliobrevin had similar inhibitory effects on the flagellar beating of demembranated and reactivated sperm and the sliding disintegration of trypsin-treated axonemes. We also analyzed the curvature and shear angle of the beating flagella and found that the proximal region of the sperm flagellum was less sensitive to ciliobrevin compared with more distal regions, where bending motions were blocked completely. Interestingly, the shear angle analysis of flagellar motility showed that ciliobrevin induced highly asymmetric bends in the proximal region of the flagellum. These results suggest that there is heterogeneity in the inhibitory thresholds of dynein motors, which depend on the regions along the flagellar shaft (proximal or distal) and on the sites of doublets in the flagellar cross-section (doublet numbers). We expect that it will be possible to map the functional differences in dynein subtypes along and/or around the cross-sections of flagellar axonemes by analyzing the inhibitory effects of ciliobrevin. (c) 2015 Wiley Periodicals, Inc.

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  • Chemotactic response with a constant delay-time mechanism in Ciona spermatozoa revealed by a high time resolution analysis of flagellar motility

    Daisuke Miyashiro, Kogiku Shiba, Tahahiro Miyashita, Shoji A. Baba, Manabu Yoshida, Shinji Kamimura

    BIOLOGY OPEN   4 ( 2 )   109 - 118   2015.2

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    During their chemotactic swimming toward eggs, sperm cells detect their species-specific chemoattractant and sense concentration gradients by unknown mechanisms. After sensing the attractant, sperm cells commonly demonstrate a series of responses involving different swimming patterns by changing flagellar beats, gradually approaching a swimming path toward the eggs, which is the source of chemoattractants. Shiba et al. observed a rapid increase in intracellular Ca2+ concentrations in Ciona spermatozoa after sensing chemoattractants; however, the biochemical processes occurring inside the sperm cells are unclear. In the present study, we focused on the timing and sensing mechanism of chemical signal detection in Ciona. One of the most crucial problems to be solved is defining the initial epoch of chemotactic responses. We adopted a high rate of video recording (600 Hz) for detailed analysis of sperm motion and a novel method for detecting subtle signs of beat forms and moving paths of sperm heads. From these analyses, we estimated a virtual sensing point of the attractant before initiation of motility responses and found that the time delay from sensing to motility responses was almost constant. To evaluate the efficiency of this constant delay model, we performed computer simulation of chemotactic behaviors of Ciona spermatozoa.

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  • Chemotactic response with a constant delay-time mechanism in Ciona spermatozoa revealed by a high time resolution analysis of flagellar motility. International journal

    Daisuke Miyashiro, Kogiku Shiba, Tahahiro Miyashita, Shoji A Baba, Manabu Yoshida, Shinji Kamimura

    Biology open   4 ( 2 )   109 - 18   2015.1

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    During their chemotactic swimming toward eggs, sperm cells detect their species-specific chemoattractant and sense concentration gradients by unknown mechanisms. After sensing the attractant, sperm cells commonly demonstrate a series of responses involving different swimming patterns by changing flagellar beats, gradually approaching a swimming path toward the eggs, which is the source of chemoattractants. Shiba et al. observed a rapid increase in intracellular Ca2+ concentrations in Ciona spermatozoa after sensing chemoattractants; however, the biochemical processes occurring inside the sperm cells are unclear. In the present study, we focused on the timing and sensing mechanism of chemical signal detection in Ciona. One of the most crucial problems to be solved is defining the initial epoch of chemotactic responses. We adopted a high rate of video recording (600 Hz) for detailed analysis of sperm motion and a novel method for detecting subtle signs of beat forms and moving paths of sperm heads. From these analyses, we estimated a virtual sensing point of the attractant before initiation of motility responses and found that the time delay from sensing to motility responses was almost constant. To evaluate the efficiency of this constant delay model, we performed computer simulation of chemotactic behaviors of Ciona spermatozoa.

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  • Choice of Prey Body Parts for Effective Feeding by Predaceous Diving Beetle Larvae, Dytiscus sharpi sharpi (Wehncke) (Coleoptera: Dytiscidae)

    Toshio Inoda, Shinji Kamimura

    JOURNAL OF INSECT BEHAVIOR   28 ( 1 )   26 - 36   2015.1

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    Diving beetle larvae use their mandibles in two ways: capturing prey and sucking their body fluid. Catching and consuming the prey's most nutritious body part leads to the highest feeding efficiency. To test this, Dytiscus sharpi sharpi larvae were given tadpoles (Rana ornativentris) as food and their feeding behaviors were observed. Dytiscus larvae preferred to catch tadpoles by the abdomens rather than by other parts. Tadpoles soon became immobilized and in most cases the beetle larvae started eating abdomens first. Beetle larvae tried to change biting site to tadpole's abdomen when the tadpole was initially caught by the head or tail. More food was absorbed from the abdomen than the head or tail suggesting that the feeding behavior of beetle larva is optimized to obtain nutrition efficiently from the tadpole abdomen.

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  • Choice of Prey Body Parts for Effective Feeding by Predaceous Diving Beetle Larvae, Dytiscus sharpi sharpi (Wehncke) (Coleoptera: Dytiscidae)

    Toshio Inoda, Shinji Kamimura

    JOURNAL OF INSECT BEHAVIOR   28 ( 1 )   26 - 36   2015.1

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    Diving beetle larvae use their mandibles in two ways: capturing prey and sucking their body fluid. Catching and consuming the prey's most nutritious body part leads to the highest feeding efficiency. To test this, Dytiscus sharpi sharpi larvae were given tadpoles (Rana ornativentris) as food and their feeding behaviors were observed. Dytiscus larvae preferred to catch tadpoles by the abdomens rather than by other parts. Tadpoles soon became immobilized and in most cases the beetle larvae started eating abdomens first. Beetle larvae tried to change biting site to tadpole's abdomen when the tadpole was initially caught by the head or tail. More food was absorbed from the abdomen than the head or tail suggesting that the feeding behavior of beetle larva is optimized to obtain nutrition efficiently from the tadpole abdomen.

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  • Dance of the microswimmers

    28 ( 10 )   30 - 37   2013.10

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  • Dance of the microswimmers

    Shinji Kamimura, riginal article, y Eric Lauga, Raymo, E. Goldstei

    Parity   28 ( 10 )   30 - 37   2013.10

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    From gut-inhabiting bacteria to sea-dwelling algae, microorganisms display a penchant for|rn|coordinated movement. Nonlinear dynamics and fluid mechanics can help explain the curious behavior.

    DOI: 10.1063/PT.3.1715

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  • X-ray diffraction recording from single axonemes of eukaryotic flagella. International journal

    Masaya Nishiura, Shiori Toba, Daisuke Takao, Daisuke Miyashiro, Hitoshi Sakakibara, Tatsuhito Matsuo, Shinji Kamimura, Kazuhiro Oiwa, Naoto Yagi, Hiroyuki Iwamoto

    Journal of structural biology   178 ( 3 )   329 - 37   2012.6

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    We report the first X-ray diffraction patterns recorded from single axonemes of eukaryotic flagella with a diameter of only <0.2 mu m, by using the technique of cryomicrodiffraction. A spermatozoon isolated from the testis of a fruit fly, Drosophila melanogaster, either intact or demembranated, was mounted straight in a glass capillary, quickly frozen and its 800-mu m segment was irradiated end-on with intense synchrotron radiation X-ray microbeams (diameter, similar to 2 mu m) at 74 K. Well-defined diffraction patterns were recorded, consisting of a large number of isolated reflection spots, extending up to 1/5 nm(-1). These reflections showed a tendency to peak every 20 degrees, i.e., the patterns had features of an 18-fold rotational symmetry as expected from the 9-fold rotational symmetry of axonemal structure. This means that the axonemes remain untwisted, even after the manual mounting procedure. The diffraction patterns were compared with the results of model calculations based on a published electron micrograph of the Drosophila axoneme. The comparison provided information about the native state of axoneme, including estimates of axonemal diameter, interdoublet spacing, and masses of axonemal components relative to those of microtubules (e.g., radial spokes, dynein arms, and proteins associated with accessory singlet microtubules). When combined with the genetic resource of Drosophila, the technique presented here will serve as a powerful tool for studying the structure-function relationship of eukaryotic flagella in general. (c) 2012 Elsevier Inc. All rights reserved.

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  • X-ray diffraction recording from single axonemes of eukaryotic flagella

    Nishiura, M., Toba, S., Takao, D., Miyashiro, D., Sakakibara, H., Matsuo, T., Kamimura, S., Oiwa, K., Yagi, N., Iwamoto, H.

    Journal of Structural Biology   178 ( 3 )   329 - 337   2012.6

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    We report the first X-ray diffraction patterns recorded from single axonemes of eukaryotic flagella with a diameter of only <0.2 mu m, by using the technique of cryomicrodiffraction. A spermatozoon isolated from the testis of a fruit fly, Drosophila melanogaster, either intact or demembranated, was mounted straight in a glass capillary, quickly frozen and its 800-mu m segment was irradiated end-on with intense synchrotron radiation X-ray microbeams (diameter, similar to 2 mu m) at 74 K. Well-defined diffraction patterns were recorded, consisting of a large number of isolated reflection spots, extending up to 1/5 nm(-1). These reflections showed a tendency to peak every 20 degrees, i.e., the patterns had features of an 18-fold rotational symmetry as expected from the 9-fold rotational symmetry of axonemal structure. This means that the axonemes remain untwisted, even after the manual mounting procedure. The diffraction patterns were compared with the results of model calculations based on a published electron micrograph of the Drosophila axoneme. The comparison provided information about the native state of axoneme, including estimates of axonemal diameter, interdoublet spacing, and masses of axonemal components relative to those of microtubules (e.g., radial spokes, dynein arms, and proteins associated with accessory singlet microtubules). When combined with the genetic resource of Drosophila, the technique presented here will serve as a powerful tool for studying the structure-function relationship of eukaryotic flagella in general. (c) 2012 Elsevier Inc. All rights reserved.

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  • Comparative structural analysis of eukaryotic flagella and cilia from Chlamydomonas, Tetrahymena, and sea urchins. International journal

    Gaia Pigino, Aditi Maheshwari, Khanh Huy Bui, Chikako Shingyoji, Shinji Kamimura, Takashi Ishikawa

    Journal of structural biology   178 ( 2 )   199 - 206   2012.5

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    Although eukaryotic flagella and cilia all share the basic 9 + 2 microtubule-organization of their internal axonemes, and are capable of generating bending-motion, the waveforms, amplitudes, and velocities of the bending-motions are quite diverse. To explore the structural basis of this functional diversity of flagella and cilia, we here compare the axonemal structure of three different organisms with widely divergent bending-motions by electron cryo-tomography. We reconstruct the 3D structure of the axoneme of Tetrahymena cilia, and compare it with the axoneme of the flagellum of sea urchin sperm, as well as with the axoneme of Chlamydomonas flagella, which we analyzed previously. This comparative structural analysis defines the diversity of molecular architectures in these organisms, and forms the basis for future correlation with their different bending-motions. (C) 2012 Elsevier Inc. All rights reserved.

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  • Comparative structural analysis of eukaryotic flagella and cilia from Chlamydomonas, Tetrahymena, and sea urchins

    Pigino, G., Maheshwari, A., Bui, K.H., Shingyoji, C., Kamimura, S., Ishikawa, T.

    Journal of Structural Biology   178 ( 2 )   199 - 206   2012.5

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    Although eukaryotic flagella and cilia all share the basic 9 + 2 microtubule-organization of their internal axonemes, and are capable of generating bending-motion, the waveforms, amplitudes, and velocities of the bending-motions are quite diverse. To explore the structural basis of this functional diversity of flagella and cilia, we here compare the axonemal structure of three different organisms with widely divergent bending-motions by electron cryo-tomography. We reconstruct the 3D structure of the axoneme of Tetrahymena cilia, and compare it with the axoneme of the flagellum of sea urchin sperm, as well as with the axoneme of Chlamydomonas flagella, which we analyzed previously. This comparative structural analysis defines the diversity of molecular architectures in these organisms, and forms the basis for future correlation with their different bending-motions. (C) 2012 Elsevier Inc. All rights reserved.

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  • 微生物のべん毛・繊毛の運動メカニズム

    上村慎治

    バイオメカニズム学会誌   34 ( 3 )   176 - 182   2010.8

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    DOI: 10.3951/sobim.34.176

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  • 微生物の運動

    上村慎治, 曲山幸生, 後藤知伸

    「生きものに学ぶ 泳ぎと飛行のしくみ」   2010.5

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  • Geometry-specific heterogeneity of the apparent diffusion rate of materials inside sperm cells. International journal

    Daisuke Takao, Shinji Kamimura

    Biophysical journal   98 ( 8 )   1582 - 8   2010.4

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    In sea urchin spermatozoa, the energy source powering flagellar motion is provided as ATP produced by mitochondria located at the proximal ends of flagella. However, the bottleneck structure between the sperm head and the flagellar tail seems to restrict the free entry of ATP from mitochondria into the tail region. To test this possibility, we investigated the diffusion properties in sperm cells using fluorescence recovery after photobleaching. We found that the rate of fluorescence recovery in the head region was similar to 10% of that observed in the flagellar tail regions. We also found that, even within the tail region, rates varied depending on location, i.e., rates were slower at the more distal regions. Using computational analysis, the rate heterogeneity was shown to be caused mainly by the geometry of the sperm structure, even if little or no difference in diffusion rates through the neck region was assumed. Therefore, we concluded that materials such as ATP would generally diffuse freely between the heads and the flagella of sperm cells. We believe these findings regarding the diffusion properties inside spermatozoa provide further insights into material transportation and chemical signaling inside eukaryotic cilia and flagella.

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  • Geometry-specific heterogeneity of the apparent diffusion rate of materials inside sperm cells

    Takao, D., Kamimura, S.

    Biophysical Journal   98 ( 8 )   1582 - 1588   2010.4

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    In sea urchin spermatozoa, the energy source powering flagellar motion is provided as ATP produced by mitochondria located at the proximal ends of flagella. However, the bottleneck structure between the sperm head and the flagellar tail seems to restrict the free entry of ATP from mitochondria into the tail region. To test this possibility, we investigated the diffusion properties in sperm cells using fluorescence recovery after photobleaching. We found that the rate of fluorescence recovery in the head region was similar to 10% of that observed in the flagellar tail regions. We also found that, even within the tail region, rates varied depending on location, i.e., rates were slower at the more distal regions. Using computational analysis, the rate heterogeneity was shown to be caused mainly by the geometry of the sperm structure, even if little or no difference in diffusion rates through the neck region was assumed. Therefore, we concluded that materials such as ATP would generally diffuse freely between the heads and the flagella of sperm cells. We believe these findings regarding the diffusion properties inside spermatozoa provide further insights into material transportation and chemical signaling inside eukaryotic cilia and flagella.

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  • Single-cell electroporation of fluorescent probes into sea urchin sperm cells and subsequent FRAP analysis Reviewed

    Takao, D., Kamimura, S.

    Zoological Science   27 ( 3 )   279 - 284   2010.3

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    In sea urchin spermatozoa, the energy required for flagellar motility depends only on the diffusional supply from proximal mitochondria, and thus the diffusion rate inside flagella is one of the most crucial factors limiting the practical size and design of the motile machinery. To determine the diffusion rates of materials inside sperm cells, FRAP (fluorescence recovery after photobleaching) analysis of incorporated fluorescent probes is one of the most powerful approaches. However, the only practically possible method until now was to use the ester forms of fluorescence, and our choice was limited to those of relatively small molecular masses, such as fluorescein derivatives. In this report, we show that a modified single-cell electroporation technique can be applied as a new microinjection method for sperm cells of the sea urchin. The method was applied to FRAP analysis to determine the rate of intraflagellar diffusion.

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  • Single-cell electroporation of fluorescent probes into sea urchin sperm cells and subsequent FRAP analysis

    Daisuke Takao, Shinji Kamimura

    Zoological Science   27 ( 3 )   279 - 284   2010.3

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    In sea urchin spermatozoa, the energy required for flagellar motility depends only on the diffusional supply from proximal mitochondria, and thus the diffusion rate inside flagella is one of the most crucial factors limiting the practical size and design of the motile machinery. To determine the diffusion rates of materials inside sperm cells, FRAP (fluorescence recovery after photobleaching) analysis of incorporated fluorescent probes is one of the most powerful approaches. However, the only practically possible method until now was to use the ester forms of fluorescence, and our choice was limited to those of relatively small molecular masses, such as fluorescein derivatives. In this report, we show that a modified single-cell electroporation technique can be applied as a new microinjection method for sperm cells of the sea urchin. The method was applied to FRAP analysis to determine the rate of intraflagellar diffusion.

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  • Structural Changes of Microtubules During GTP Hydrolysis Revealed by X-Ray Fiber-Diffraction

    Shinji Kamimura, Hiroyuki Iwamoto, Daisuke Miyashiro

    BIOPHYSICAL JOURNAL   98 ( 3 )   361A - 361A   2010.1

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  • Quick shear-flow alignment of biological filaments for X-ray fiber diffraction facilitated by methylcellulose

    Sugiyama, T., Miyashiro, D., Takao, D., Iwamoto, H., Sugimoto, Y., Wakabayashi, K., Kamimura, S.

    Biophysical Journal   97 ( 12 )   3132 - 3138   2009.12

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    X-ray fiber diffraction is one of the most useful methods for examining the structural details of live biological filaments under physiological conditions. To investigate biologically active or labile materials, it is crucial to finish fiber alignment within seconds before diffraction analysis. However, the conventional methods, e.g., magnetic field alignment and low-speed centrifugations, are time-consuming and not very useful for such purposes. Here, we introduce a new alignment method using a rheometer with two parallel disks, which was applied to observe fiber diffractions of axonemes, tobacco mosaic tobamovirus, and microtubules. We found that fibers were aligned within 5 s by giving high shearflow (1000-5000 s(-1)) to the medium and that methylcellulose contained in the medium (similar to 1%) was essential to the accomplishment of uniform orientation with a small angular deviation (<5 degrees). The new alignment method enabled us to execute structure analyses of axonemes by small-angle x-ray diffraction. Since this method was also useful for the quick alignment of purified microtubules, as well as tobacco mosaic tobamovirus, we expect that we can apply it to the structural analysis of many other biological filaments.

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  • Quick shear-flow alignment of biological filaments for X-ray fiber diffraction facilitated by methylcellulose. International journal

    Takaaki Sugiyama, Daisuke Miyashiro, Daisuke Takao, Hiroyuki Iwamoto, Yasunobu Sugimoto, Katsuzo Wakabayashi, Shinji Kamimura

    Biophysical journal   97 ( 12 )   3132 - 8   2009.12

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    X-ray fiber diffraction is one of the most useful methods for examining the structural details of live biological filaments under physiological conditions. To investigate biologically active or labile materials, it is crucial to finish fiber alignment within seconds before diffraction analysis. However, the conventional methods, e.g., magnetic field alignment and low-speed centrifugations, are time-consuming and not very useful for such purposes. Here, we introduce a new alignment method using a rheometer with two parallel disks, which was applied to observe fiber diffractions of axonemes, tobacco mosaic tobamovirus, and microtubules. We found that fibers were aligned within 5 s by giving high shearflow (1000-5000 s(-1)) to the medium and that methylcellulose contained in the medium (similar to 1%) was essential to the accomplishment of uniform orientation with a small angular deviation (<5 degrees). The new alignment method enabled us to execute structure analyses of axonemes by small-angle x-ray diffraction. Since this method was also useful for the quick alignment of purified microtubules, as well as tobacco mosaic tobamovirus, we expect that we can apply it to the structural analysis of many other biological filaments.

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  • Dietary program for rearing the larvae of a diving beetle, Dytiscus sharpi (Wehncke), in the laboratory (Coleoptera: Dytiscidae)

    Inoda, T., Hasegawa, M., Kamimura, S., Hori, M.

    Coleopterists Bulletin   63 ( 3 )   340 - 350   2009.9

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    For the conservation of the diving beetle Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae), which is included on the Red Data List of Japan, it is critical to understand its ecological background. In the present study, we focused on its feeding behavior and nutritional needs under laboratory breeding conditions. First, we made a list of the possible candidates of prey in the same habitats where we caught D. sharpi. We found that the tadpoles of Rana ornativentris (Werner) were the major species present from March to April, when the beetle larvae appeared. Second, under our laboratory conditions, we investigated the size preference of beetle larvae preying on R. ornativentris tadpoles. We found a significant positive correlation between the developing stage of the larvae and the preferred prey size, i.e., the first and third instars preferred smaller and larger prey, respectively, but second instars did not show any size preference. The size of full-grown adult beetles was almost the same as that of wild insects found in the field, indicating that R. ornativentris tadpoles provide almost complete nutrition for larval growth. Finally, we investigated how the size and number of R. ornativentris tadpoles were correlated with the developing stage of beetle larvae. We suggest that it is crucial for Dytiscus larvae to have access to tadpoles of the proper size and amounts, depending on their growth stage. © 2009 Coleopterists Society.

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  • DIETARY PROGRAM FOR REARING THE LARVAE OF A DIVING BEETLE, DYTISCUS SHARPI (WEHNCKE), IN THE LABORATORY (COLEOPTERA: DYTISCIDAE)

    Toshio Inoda, Masami Hasegawa, Shinji Kamimura, Michio Hori

    COLEOPTERISTS BULLETIN   63 ( 3 )   340 - 350   2009.9

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    For the conservation of the diving beetle Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae), which is included on the Red Data List of Japan, it is critical to understand its ecological background. In the present study, we focused on its feeding behavior and nutritional needs under laboratory breeding conditions. First, we made a list of the possible candidates of prey in the same habitats where we caught D. sharpi. We found that the tadpoles of Rana ornativentris (Werner) were the major species present from March to April, when the beetle larvae appeared. Second, under our laboratory conditions, we investigated the size preference of beetle larvae preying on R. ornativentris tadpoles. We found a significant positive correlation between the developing stage of the larvae and the preferred prey size, i.e., the first and third instars preferred smaller and larger prey, respectively, but second instars did not show any size preference. The size of full-grown adult beetles was almost the same as that of wild insects found in the field, indicating that R. ornativentris tadpoles provide almost complete nutrition for larval growth. Finally, we investigated how the size and number of R. ornativentris tadpoles were correlated with the developing stage of beetle larvae. We suggest that it is crucial for Dytiscus larvae to have access to tadpoles of the proper size and amounts, depending on their growth stage. © 2009 Coleopterists Society.

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  • X-ray fiber diffraction studies on flagellar axonemes.

    Oiwa, K., Kamimura, S., Iwamoto, H.

    Methods in cell biology   91 ( 5 )   89 - 109   2009

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    Eukaryotic cilia and flagella are highly ordered and precisely assembled cellular organelles. Here, to understand the mechanism of the orderly undulations of cilia and flagella, we shall draw a blueprint of their core structures and supporting scaffolds, that is, axonemes, and we shall describe the dynamic structural changes of components of the organelles. Small-angle X-ray scattering and diffraction are among the principal tools used to study protein polymers. These methods are now well established as indispensable tools that complement electron microscopy, providing information on the structure and dynamics of biological materials at atomic resolution in near-physiological environments. For instance, X-ray diffraction studies of skeletal muscles have contributed greatly to our understanding of the structure and molecular mechanisms of muscles. However, owing to the minute size and low diffracting power of axonemes, few attempts at X-ray diffraction of axonemes have been reported. The advent of third-generation synchrotron radiation facilities now makes these attempts feasible, because we now have stable and intense X-rays that enable us to obtain diffractions from the axonemes. In this chapter, we provide a concise practical guide to this new avenue for structural analysis of axonemes.

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  • X-ray fiber diffraction studies on flagellar axonemes. International journal

    Kazuhiro Oiwa, Shinji Kamimura, Hiroyuki Iwamoto

    Methods in cell biology   91   89 - 109   2009

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    Eukaryotic cilia and flagella are highly ordered and precisely assembled cellular organelles. Here, to understand the mechanism of the orderly undulations of cilia and flagella, we shall draw a blueprint of their core structures and supporting scaffolds, that is, axonemes, and we shall describe the dynamic structural changes of components of the organelles. Small-angle X-ray scattering and diffraction are among the principal tools used to study protein polymers. These methods are now well established as indispensable tools that complement electron microscopy, providing information on the structure and dynamics of biological materials at atomic resolution in near-physiological environments. For instance, X-ray diffraction studies of skeletal muscles have contributed greatly to our understanding of the structure and molecular mechanisms of muscles. However, owing to the minute size and low diffracting power of axonemes, few attempts at X-ray diffraction of axonemes have been reported. The advent of third-generation synchrotron radiation facilities now makes these attempts feasible, because we now have stable and intense X-rays that enable us to obtain diffractions from the axonemes. In this chapter, we provide a concise practical guide to this new avenue for structural analysis of axonemes.

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  • FRAP analysis of molecular diffusion inside sea-urchin spermatozoa

    Daisuke Takao, Shinji Kamimura

    Journal of Experimental Biology   211 ( 22 )   3594 - 3600   2008.11

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    In sea-urchin spermatozoa, energy required for flagellar motility is provided by ATP diffusion from mitochondria located at the proximal ends of the flagella along with the creatine shuttle system. However, no direct analysis of the diffusion rates inside flagella has been carried out thus far. Using a FRAP (fluorescence recovery after photobleaching) technique, we determined the diffusion coefficients of fluorescein-derivatives (calcein, carboxyfluorescein and Oregon Green) to be 63-64 mu m(2) s(-1). Although these values are about one third of the estimates that were previously used for theoretical calculations, we concluded that the rate of ATP diffusion inside spermatozoa was high enough to support the continuous motility of sea-urchin sperm flagella if the creatine shuttle system is working. We also investigated the diffusion rate through the 'neck' region between the head and tail. When the head region of a calcein-loaded spermatozoon was photobleached, slow recovery of head fluorescence along with the decrease of fluorescence signal in the tail region was observed. It suggests that small molecules such as calcein (M(r), 622.54) can move beyond the boundary between the head and the flagellum. We expect that these findings regarding the diffusion properties inside spermatozoa will provide us with more general insights into the energy equilibrium and material transportation by passive diffusion inside eukaryotic cilia and flagella.

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  • FRAP analysis of molecular diffusion inside sea-urchin spermatozoa International journal

    Takao, D., Kamimura, S.

    Journal of Experimental Biology   211 ( 22 )   3594 - 600   2008.11

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    In sea-urchin spermatozoa, energy required for flagellar motility is provided by ATP diffusion from mitochondria located at the proximal ends of the flagella along with the creatine shuttle system. However, no direct analysis of the diffusion rates inside flagella has been carried out thus far. Using a FRAP (fluorescence recovery after photobleaching) technique, we determined the diffusion coefficients of fluorescein-derivatives (calcein, carboxyfluorescein and Oregon Green) to be 63-64μm2s-1. Although these values are about one third of the estimates that were previously used for theoretical calculations, we concluded that the rate of ATP diffusion inside spermatozoa was high enough to support the continuous motility of sea-urchin sperm flagella if the creatine shuttle system is working. We also investigated the diffusion rate through the 'neck' region between the head and tail. When the head region of a calcein-loaded spermatozoon was photobleached, slow recovery of head fluorescence along with the decrease of fluorescence signal in the tail region was observed. It suggests that small molecules such as calcein (M r, 622.54) can move beyond the boundary between the head and the flagellum. We expect that these findings regarding the diffusion properties inside spermatozoa will provide us with more general insights into the energy equilibrium and material transportation by passive diffusion inside eukaryotic cilia and flagella.

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  • Temperature-dependent regulation of reproduction in the Diving Beetle Dytiscus sharpi (Coleoptera: Dytiscidae)

    Inoda, T., Tajima, F., Taniguchi, H., Saeki, M., Numakura, K., Hasegawa, M., Kamimura, S.

    Zoological Science   24 ( 11 )   1115 - 21   2007.11

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    The effects of temperature on the mating behavior, gonad development, germ cell maturation, and egg spawning of the predaceous diving beetle Dytiscus sharpi (Coleoptera; Dytiscidae), were investigated. By field observations, we found that mating behavior started in October and occurred more frequently from November to December. Under our laboratory breeding conditions, we observed almost the same seasonal variation in mating behavior. We found that temperatures lower than 20 degrees C were required to trigger mating behavior. We also found the same temperature threshold triggered gonadogenesis as well as spermatogenesis. Furthermore, for females, exposure to lower temperatures (<8 degrees C) during the winter was required for egg maturation and spawning in spring; that is, there was a second threshold for successful female reproduction. We conclude that the termination of summer reproductive diapause of D. sharpi is regulated in a temperature-dependent manner, thus effecting the adaptation of D. sharpi to southern warm habitats.

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  • Temperature-dependent regulation of reproduction in the Diving Beetle Dytiscus sharpi (Coleoptera: Dytiscidae)

    Toshio Inoda, Fumitada Tajima, Hiroshi Taniguchi, Motoyuki Saeki, Kazuki Numakura, Masami Hasegawa, Shinji Kamimura

    Zoological Science   24 ( 11 )   1115 - 1121   2007.11

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    The effects of temperature on the mating behavior, gonad development, germ cell maturation, and egg spawning of the predaceous diving beetle Dytiscus sharpi (Coleoptera; Dytiscidae), were investigated. By field observations, we found that mating behavior started in October and occurred more frequently from November to December. Under our laboratory breeding conditions, we observed almost the same seasonal variation in mating behavior. We found that temperatures lower than 20 degrees C were required to trigger mating behavior. We also found the same temperature threshold triggered gonadogenesis as well as spermatogenesis. Furthermore, for females, exposure to lower temperatures (<8 degrees C) during the winter was required for egg maturation and spawning in spring; that is, there was a second threshold for successful female reproduction. We conclude that the termination of summer reproductive diapause of D. sharpi is regulated in a temperature-dependent manner, thus effecting the adaptation of D. sharpi to southern warm habitats.

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  • 動物個体の環境応答と情報処理

    上村慎治

    生命科学   239 - 251   2007.2

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  • Analysis of small-angle X-ray diffractions from the flow-aligned axonemes of sea-urchin spermatozoa.

    Shinji Kamimura, Hiroyuki Lwamoto, Tetsuro Fujisawa

    BIOPHYSICAL JOURNAL   500A - 500A   2007.1

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  • 空と海の生物に学ぶ ―バイオメカニクスの挑戦―

    上村慎治

    遺伝   61 ( 61 )   30 - 32   2007.1

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  • 基礎生命科学実験

    上村慎治

    ( i-ii )   184 - 186   2007.1

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  • Dynamic flow alignment of flagellar axonemes for low-angle X-ray fiber diffraction analysis.

    Kamimura, S, Sugiyama, T, Sugimoto, Y, Walabayashi, K

    Photon Factory Actuivity Report 2006   2005-23 ( Part B )   2006.12

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    Kamimura, S, Sugiyama, T, Sugimoto, Y, Walabayashi, K

    Photon Factory Actuivity Report 2006   2006.12

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  • Ciliated cells differentiated from mouse embryonic stem cells. International journal

    Yusuke Nishimura, Tatsuo S Hamazaki, Shinji Komazaki, Shinji Kamimura, Hitoshi Okochi, Makoto Asashima

    Stem cells (Dayton, Ohio)   24 ( 5 )   1381 - 8   2006.5

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    In the present study, we demonstrated that the mouse embryonic stem cells were differentiated into ciliated epithelial cells, with characteristics of normal ciliated cells. These cells expressed ciliary marker proteins, such as beta-tubulin IV and hepatocyte nuclear factor-3/forkhead homolog 4 (HFH-4), and processed microtubules were arranged in the 9 + 2 structure, which is the same specific alignment observed in normal ciliary microtubules. The cilia of these cells were beating at a frequency of 17-20 Hz. The differentiated embryoid bodies (EBs) containing these ciliated cells expressed respiratory marker genes such as thyroid transcription factor-1 and surfactant protein-C. For the induction of ciliated cells, culture of EBs in serum-free medium during the initial 2 days of the attachment was indispensable. When EBs were treated with bone morphogenetic proteins, the expression of HFH-4 was decreased, and the ciliated cells were scarcely differentiated. Previous methods for inducing ciliated cells in vitro from embryonic or adult tissues involved an air-liquid interface. The system used in this study more closely mimics the normal development of ciliated cells; thus, an added advantage of the system is as a tool for studying the differentiation mechanism of normal ciliated epithelial cells.

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  • Ciliated cells differentiated from mouse embryonic stem cells

    Nishimura, Y., Hamazaki, T.S., Komazaki, S., Kamimura, S., Okochi, H., Asashimaa, M.

    Stem Cells   24 ( 5 )   1381 - 1388   2006.5

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    In the present study, we demonstrated that the mouse embryonic stem cells were differentiated into ciliated epithelial cells, with characteristics of normal ciliated cells. These cells expressed ciliary marker proteins, such as beta-tubulin IV and hepatocyte nuclear factor-3/forkhead homolog 4 (HFH-4), and processed microtubules were arranged in the 9 + 2 structure, which is the same specific alignment observed in normal ciliary microtubules. The cilia of these cells were beating at a frequency of 17-20 Hz. The differentiated embryoid bodies (EBs) containing these ciliated cells expressed respiratory marker genes such as thyroid transcription factor-1 and surfactant protein-C. For the induction of ciliated cells, culture of EBs in serum-free medium during the initial 2 days of the attachment was indispensable. When EBs were treated with bone morphogenetic proteins, the expression of HFH-4 was decreased, and the ciliated cells were scarcely differentiated. Previous methods for inducing ciliated cells in vitro from embryonic or adult tissues involved an air-liquid interface. The system used in this study more closely mimics the normal development of ciliated cells; thus, an added advantage of the system is as a tool for studying the differentiation mechanism of normal ciliated epithelial cells.

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  • Increase in intracellular pH induces phosphorylation of axonemal proteins for activation of flagellar motility in starfish sperm International journal

    Ayako Nakajima, Masaya Morita, Akihiro Takemura, Shinji Kamimura, Makoto Okuno

    Journal of Experimental Biology   208 ( 23 )   4411 - 4418   2005.12

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    Increased intracellular pH ([pH](i)) activates dynein in sea urchin and mammalian sperm and induces activation of flagellar motility. It is thought that cAMP-dependent protein phosphorylation is associated with motility activation through increasing [pH](i), but little attention has been given to the cAMP-independent phosphorylation also induced by the [pH](i) increase. The present study demonstrates that the increase in [pH](i) in starfish sperm induces the phosphorylation of axonemal proteins and activation of flagellar motility independently of cAMP. Flagellar motility of intact sperm was activated when the [pH](i) was raised by addition of NH4Cl. Histidine, which is known to activate motility of starfish sperm, also raised the [pH](i) during the motility activation. In addition, motility of demembranated sperm flagella was activated in a pH-dependent manner without cAMP. These results indicate that in starfish sperm it is the increase in [pH](i) that induces activation of flagellar motility. Moreover, phosphorylation of axonemal proteins (of molecular mass 25, 32 and 45 kDa) was observed during the pH-dependent and cAMP-independent motility activation of demembranated sperm. This suggests that the increase in [pH](i) regulates flagellar motility via cAMP-independent phosphorylation of axonemal proteins.

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  • A new ratio-imaging technique to measure intracellular pH of sea-urchin sperm flagella

    Daisuke Takao, Shinji Kamimura

    ZOOLOGICAL SCIENCE   22 ( 12 )   1447 - 1447   2005.12

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  • Increase in intracellular pH induces phosphorylation of axonemal proteins for activation of flagellar motility in starfish sperm Reviewed

    Nakajima, A., Morita, M., Takemura, A., Kamimura, S., Okuno, M.

    Journal of Experimental Biology   208 ( 23 )   4411 - 4418   2005.11

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  • Differentiation of ciliated cellsin vitro

    Y. Nishimura, T. S. Hamazaki, S. Komazaki, S. Kamimura, M. Asashima

    MECHANISMS OF DEVELOPMENT   122   S132 - S132   2005.9

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  • New open aquarium system to breed larvae of water beetles (Coleoptera: Dytiscidae)

    Toshio Inoda, Shinji Kamimura

    Coleopterists Bulletin   58 ( 1 )   37 - 43   2004.3

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    For conservation purposes and to supply rare insects for laboratory use, a system for artificial breeding is crucial. However, in the case of carnivorous freshwater insects such as diving beetles, constant conditions in aquariums are difficult to maintain due to their high rate of food consumption. Furthermore, surface rippling caused by the pumping system for water circulation hinders the respiration of small larvae. We developed a new open aquarium system without water circulation that was successfully applied to the rearing of larvae of diving beetles, Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae). In comparison to conventional methods, a high proportion of larvae developed into adult insects. The size of reared adults was almost the same as those of field-collected adults. The new method could be applied to the conservation and breeding of other rare species, such as water beetles and water bugs.

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  • New open aquarium system to breed larvae of water beetles (Coleoptera: Dytiscidae)

    Inoda, T., Kamimura, S.

    Coleopterists Bulletin   58 ( 1 )   37 - 43   2004.3

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    For conservation purposes and to supply rare insects for laboratory use, a system for artificial breeding is crucial. However, in the case of carnivorous freshwater insects such as diving beetles, constant conditions in aquariums are difficult to maintain due to their high rate of food consumption. Furthermore, surface rippling caused by the pumping system for water circulation hinders the respiration of small larvae. We developed a new open aquarium system without water circulation that was successfully applied to the rearing of larvae of diving beetles, Dytiscus sharpi (Wehncke) (Coleoptera: Dytiscidae). In comparison to conventional methods, a high proportion of larvae developed into adult insects. The size of reared adults was almost the same as those of field-collected adults. The new method could be applied to the conservation and breeding of other rare species, such as water beetles and water bugs.

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  • Diameter Oscillation of Axonemes in Sea-Urchin Sperm Flagella

    Sakakibara, H.M., Kunioka, Y., Yamada, T., Kamimura, S.

    Biophysical Journal   86 ( 1 I )   346 - 352   2004.1

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    The 9+2 configuration of axonemes is one of the most conserved structures of eukaryotic organelles. Evidence so far has confirmed that bending of cilia and flagella is the result of active sliding of microtubules induced by dynein arms. If the conformational change of dynein motors, which would be a key step of force generation, is occurring in a three-dimensional manner, we can easily expect that the microtubule sliding should contain some transverse component, i.e., a motion in a direction at a right angle to the longitudinal axis of axonemes. Using a modified technique of atomic force microscopy, we found such transverse motion is actually occurring in an oscillatory manner when the axonemes of sea-urchin sperm flagella were adhered onto glass substrates. The motion was adenosine triphosphate-dependent and the observed frequency of oscillation was similar to that of oscillatory sliding of microtubules that had been shown to reflect the physiological activity of dynein arms (S. Kamimura and R. Kamiya. 1989. Nature. 340: 476-478; 1992. J. Cell Biol. 116: 1443-1454). Maximal amplitude of the diameter oscillation was around 10 nm, which was within a range of morphological change observed with electron microscopy (F.D. Warner. 1978. J. Cell Biol. 77: R19-R26; N. C. Zanetti, D. R. Mitchell, and F. D. Warner. 1979. J. Cell Biol. 80: 573-588).

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  • Diameter Oscillation of Axonemes in Sea-Urchin Sperm Flagella

    Hajime M. Sakakibara, Yuki Kunioka, Takenori Yamada, Shinji Kamimura

    Biophysical Journal   86 ( 1 I )   346 - 352   2004.1

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    The 9+2 configuration of axonemes is one of the most conserved structures of eukaryotic organelles. Evidence so far has confirmed that bending of cilia and flagella is the result of active sliding of microtubules induced by dynein arms. If the conformational change of dynein motors, which would be a key step of force generation, is occurring in a three-dimensional manner, we can easily expect that the microtubule sliding should contain some transverse component, i.e., a motion in a direction at a right angle to the longitudinal axis of axonemes. Using a modified technique of atomic force microscopy, we found such transverse motion is actually occurring in an oscillatory manner when the axonemes of sea-urchin sperm flagella were adhered onto glass substrates. The motion was adenosine triphosphate-dependent and the observed frequency of oscillation was similar to that of oscillatory sliding of microtubules that had been shown to reflect the physiological activity of dynein arms (S. Kamimura and R. Kamiya. 1989. Nature. 340: 476-478; 1992. J. Cell Biol. 116: 1443-1454). Maximal amplitude of the diameter oscillation was around 10 nm, which was within a range of morphological change observed with electron microscopy (F.D. Warner. 1978. J. Cell Biol. 77: R19-R26; N. C. Zanetti, D. R. Mitchell, and F. D. Warner. 1979. J. Cell Biol. 80: 573-588).

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  • Asymmetric Mandibles of Water-Scavenger Larvae Improve Feeding Effectiveness on Right-Handed Snails

    Inoda, T., Hirata, Y., Kamimura, S.

    American Naturalist   162 ( 6 )   811 - 814   2003.12

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    DOI: 10.1086/378903

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  • Asymmetric Mandibles of Water-Scavenger Larvae Improve Feeding Effectiveness on Right-Handed Snails

    Toshio Inoda, Yoshiyuki Hirata, Shinji Kamimura

    American Naturalist   162 ( 6 )   811 - 814   2003.12

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    DOI: 10.1086/378903

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  • Effects of Vomeronasal Organ Removal on the Sperm Motility in Male Mice

    Koyama, S., Kamimura, S.

    Zoological Science   20 ( 11 )   1355 - 1358   2003.11

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    Odors play important roles in the communication of house mice. They release behaviors and prime changes of the physiological conditions of other individuals. In our previous study, we showed that sperm motility was lowered in the subordinate mice comparing with dominant mice. Our hypothesis is that the lowered sperm motility was due to some primer effects by odor substances derived from dominant mice. To test the hypothesis, we destroyed the vomeronasal organ (VNO) of male mice (VNX male) at 5 weeks of age and paired them with intact male mice (Experimental Group). As control group males, intact male mice were kept in pairs (Control Group). At 15 weeks of age, the sperm motility and weights of reproductive organs, and social dominance was analyzed. The subordinate VNX males were found to have high sperm motility comparable to the dominant males. It was suggested that there is male-to-male primer effects, mediated by VNO, that suppress sperm motility of the subordinate mice.

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  • Study on the development of sperm motility and social dominance of male mice

    Koyama, S., Kamimura, S.

    Physiology and Behavior   80 ( 2-3 )   267 - 272   2003.11

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    It has been shown that sperm motility and other parameters related to the reproductive activity varied depending on the social status of male mice. In order to clarify whether such variation is derived from inborn difference or depends on any conditions during maturation, we investigated developmental change of sperm motility, reproductive organs, and body size of MY male mice. We also investigated how the establishment of social dominance of male mice during maturation was correlated with sperm motility. It was found that sperm motility was significantly higher during puberty than in adulthood, although there already existed relatively large statistical variance. The correlation between sperm motility and the social status was revealed to start after 10 weeks of age. It was suggested that a certain inborn difference of sperm motility became enlarged due to environmental factors experienced by male mice during maturation. (C) 2003 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.physbeh.2003.07.001

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  • Study on the development of sperm motility and social dominance of male mice

    Sachiko Koyama, Shinji Kamimura

    Physiology and Behavior   80 ( 2-3 )   267 - 272   2003.11

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    It has been shown that sperm motility and other parameters related to the reproductive activity varied depending on the social status of male mice. In order to clarify whether such variation is derived from inborn difference or depends on any conditions during maturation, we investigated developmental change of sperm motility, reproductive organs, and body size of MY male mice. We also investigated how the establishment of social dominance of male mice during maturation was correlated with sperm motility. It was found that sperm motility was significantly higher during puberty than in adulthood, although there already existed relatively large statistical variance. The correlation between sperm motility and the social status was revealed to start after 10 weeks of age. It was suggested that a certain inborn difference of sperm motility became enlarged due to environmental factors experienced by male mice during maturation. (C) 2003 Elsevier Inc. All rights reserved.

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  • Effects of Vomeronasal Organ Removal on the Sperm Motility in Male Mice

    Sachiko Koyama, Shinji Kamimura

    Zoological Science   20 ( 11 )   1355 - 1358   2003.11

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    Odors play important roles in the communication of house mice. They release behaviors and prime changes of the physiological conditions of other individuals. In our previous study, we showed that sperm motility was lowered in the subordinate mice comparing with dominant mice. Our hypothesis is that the lowered sperm motility was due to some primer effects by odor substances derived from dominant mice. To test the hypothesis, we destroyed the vomeronasal organ (VNO) of male mice (VNX male) at 5 weeks of age and paired them with intact male mice (Experimental Group). As control group males, intact male mice were kept in pairs (Control Group). At 15 weeks of age, the sperm motility and weights of reproductive organs, and social dominance was analyzed. The subordinate VNX males were found to have high sperm motility comparable to the dominant males. It was suggested that there is male-to-male primer effects, mediated by VNO, that suppress sperm motility of the subordinate mice.

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  • 水薄膜内での精子遊泳観察 Reviewed

    上村慎治

    日本機械学会年次大会講演論文集   ( 6 )   135 - 138   2003.6

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  • Imaging of the fluorescence spectrum of a single fluorescent molecule by prism-based spectroscopy

    Yoshikazu Suzuki, Tomomi Tani, Kazuo Sutoh, Shinji Kamimura

    FEBS Letters   512 ( 1-3 )   235 - 239   2002.2

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    We have devised a novel method to visualize the fluorescence spectrum of a single fluorescent molecule using prism-based spectroscopy. Equiping a total internal reflection microscope with a newly designed wedge prism, we obtained a spectral image of a single rhodamine red molecule attached to an essential light chain of myosin. We also obtained a spectral image of single-pair fluorescence resonance energy transfer between rhodamine red and Cy5 in a double-labeled myosin motor domain. This method could become a useful tool to investigate the dynamic processes of biomolecules at the single-molecule level. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

    DOI: 10.1016/S0014-5793(02)02269-X

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  • Imaging of the fluorescence spectrum of a single fluorescent molecule by prism-based spectroscopy

    Suzuki, Y., Tani, T., Sutoh, K., Kamimura, S.

    FEBS Letters   512 ( 1-3 )   235 - 239   2002.2

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    We have devised a novel method to visualize the fluorescence spectrum of a single fluorescent molecule using prism-based spectroscopy. Equiping a total internal reflection microscope with a newly designed wedge prism, we obtained a spectral image of a single rhodamine red molecule attached to an essential light chain of myosin. We also obtained a spectral image of single-pair fluorescence resonance energy transfer between rhodamine red and Cy5 in a double-labeled myosin motor domain. This method could become a useful tool to investigate the dynamic processes of biomolecules at the single-molecule level. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

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  • Effects of social dominance and female odor on sperm activity of male mice

    S Koyama, S Kamimura

    CHEMICAL SIGNALS IN VERTEBRATES 9   9 ( 9 )   403 - 410   2001

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  • Effects of social dominance and female odor on sperm activity of male mice

    S Koyama, S Kamimura

    CHEMICAL SIGNALS IN VERTEBRATES 9   9   403 - 410   2001

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  • Influence of social dominance and female odor on the sperm activity of male mice

    Sachiko Koyama, Shinji Kamimura

    Physiology and Behavior   71 ( 3-4 )   415 - 422   2000.11

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    In mammals, sperm activity is known to be varied largely according to individuals though physiological reasons have not been clarified yet. In our previous study [Koyama S, Kamimura S. Lowered sperm motility in mice of subordinate social status. Physiol Behav 1999;65:665-669.], we showed that sperm motility was higher in the dominant mice than the subordinate mice, by which it was suggested that social factors could affect sperm activity in mammals. In the present study, we investigated how the observed influence of social dominance would be modified by the existence of females. From 5 to 15 weeks of age, male mice were pair housed and were kept under three different housing conditions: (1) with females; (2) with bedding soiled by females; and (3) control group. The social dominance of the paired males was determined by resident-intruder tests that were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of organs, level of serum testosterone and corticosterone were determined. It was revealed that sperm density was higher and weight of preputial glands was heavier in dominants than in subordinates when they were kept with females or female bedding, In the subordinates, however, there were no differences among the three housing conditions; that is, there were no female effects on the subordinates. On the other hand, sperm motility was high in the dominants of control group, low in the subordinates, and lower in the dominants that were kept with females. The dominants of the males that were kept with females showed high aggressiveness, and there were negative correlationships to be seen between aggressiveness and sperm motility. It was suggested that: (1) Female odor promotes spermatogenesis of the dominants, but it does not promote that of the subordinates. (2) Sperm motility is more affected by social dominance than by female odor. (3) Excessive aggressiveness has negative influence on sperm motility. (C) 2000 Elsevier Science Inc. All rights reserved.

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  • Influence of social dominance and female odor on the sperm activity of male mice

    Koyama, S., Kamimura, S.

    Physiology and Behavior   71 ( 3-4 )   415 - 422   2000.11

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    In mammals, sperm activity is known to be varied largely according to individuals though physiological reasons have not been clarified yet. In our previous study [Koyama S, Kamimura S. Lowered sperm motility in mice of subordinate social status. Physiol Behav 1999;65:665-669.], we showed that sperm motility was higher in the dominant mice than the subordinate mice, by which it was suggested that social factors could affect sperm activity in mammals. In the present study, we investigated how the observed influence of social dominance would be modified by the existence of females. From 5 to 15 weeks of age, male mice were pair housed and were kept under three different housing conditions: (1) with females; (2) with bedding soiled by females; and (3) control group. The social dominance of the paired males was determined by resident-intruder tests that were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of organs, level of serum testosterone and corticosterone were determined. It was revealed that sperm density was higher and weight of preputial glands was heavier in dominants than in subordinates when they were kept with females or female bedding, In the subordinates, however, there were no differences among the three housing conditions; that is, there were no female effects on the subordinates. On the other hand, sperm motility was high in the dominants of control group, low in the subordinates, and lower in the dominants that were kept with females. The dominants of the males that were kept with females showed high aggressiveness, and there were negative correlationships to be seen between aggressiveness and sperm motility. It was suggested that: (1) Female odor promotes spermatogenesis of the dominants, but it does not promote that of the subordinates. (2) Sperm motility is more affected by social dominance than by female odor. (3) Excessive aggressiveness has negative influence on sperm motility. (C) 2000 Elsevier Science Inc. All rights reserved.

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  • Dynein-ADP as a force generating intermediate revealed by the rapid reactivation of microtubule sliding in sand-dollar sperm flagella after the photolysis of caged ATP Reviewed

    Tani, T, Kamimura, S

    Biophys. J.   ( 77 )   1518 - 1527   1999.9

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    DOI: 10.1016/S0006-3495(99)76999-7

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  • Dynein-ADP as a force generating intermediate revealed by the rapid reactivation of microtubule sliding in sand-dollar sperm flagella after the photolysis of caged ATP

    Tani, T, Kamimura, S

    Biophys. J.   ( 77 )   1518 - 1527   1999.9

  • 生物光学顕微鏡の基礎

    上村慎治

    生命科学を拓く新しい光技術 (シリーズ・光が拓く生命科学)   28 - 46   1999.4

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  • CHEMO-MECHAMCAL COUPLING OF SLIDING BY AXONEMAL DYNEIN(Physiology)(Proceedings of the Seventieth Annual Meeting of the Zoological Society of Japan) :

    Kamimura S., Tani T.

    Zoological science   16   96 - 96   1999

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  • Dynein-ADP as a force generating intermediate revealed by the reactivation of sand-dollar spera flagella after the photolysis of caged ATP.

    Tani T., Kamimura S.

    Biophysics   38 ( 2 )   S73   1998.9

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  • Reactivation of sea-urchin sperm flagella induced by rapid photolysis of caged ATP

    Tani, T., Kamimura, S.

    Journal of Experimental Biology   201 ( 10 )   1493 - 1503   1998.5

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    Sea-urchin sperm flagella in a state of rigor were reactivated by rapid photolysis of caged ATP, After a time lag of 11-17 ms, all bends in the axonemes present during rigor began to be propagated towards the tip as if their propagation had not been interrupted. This result suggests that the site-specific activity of dyneins along the length of the axoneme is preserved even during rigor states when ATP is absent and that regulation of the activity can be restarted immediately with a new cycle of ATP turnover. During the starting transient, pre-existing rigor waves in the distal region were propagated without a change in the maximal shear angle until they disappeared at the tip. This was more evident when the rapid reactivation was triggered in high-viscosity solution, in which only the form of new bends was greatly affected by viscous load. After reactivation, the velocity of microtubule sliding increased and reached a plateau within 28 ms. This time course reflects the rate of force generation by dynein in situ.

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  • Reactivation of sea-urchin sperm flagella induced by rapid photolysis of caged ATP

    Tomomi Tani, Shinji Kamimura

    Journal of Experimental Biology   201 ( 10 )   1493 - 1503   1998.5

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    Sea-urchin sperm flagella in a state of rigor were reactivated by rapid photolysis of caged ATP, After a time lag of 11-17 ms, all bends in the axonemes present during rigor began to be propagated towards the tip as if their propagation had not been interrupted. This result suggests that the site-specific activity of dyneins along the length of the axoneme is preserved even during rigor states when ATP is absent and that regulation of the activity can be restarted immediately with a new cycle of ATP turnover. During the starting transient, pre-existing rigor waves in the distal region were propagated without a change in the maximal shear angle until they disappeared at the tip. This was more evident when the rapid reactivation was triggered in high-viscosity solution, in which only the form of new bends was greatly affected by viscous load. After reactivation, the velocity of microtubule sliding increased and reached a plateau within 28 ms. This time course reflects the rate of force generation by dynein in situ.

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  • 光学顕微鏡入門 - 虫眼鏡から超顕微鏡へ

    上村慎治

    バイオイメージング (シリーズ・ニューバイオフィジックス)   29 - 48   1998.4

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  • CHEMO-MECHANICAL COUPLING AND FORCE-GENERATING INTERMEDIATE OF AXONEMAL DYNEIN REVEALED BY A RAPID REACTIVATION EXPERIMENT WITH CAGED-ATP(Physiology)(Proceedings of the Sixty-Ninth Annual Meeting of the Zoological Society of Japan) :

    Tani T., Kamimura S.

    Zoological science   15   100 - 100   1998

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  • Lowered sperm motility in subordinate social status of mice

    Sachiko Koyama, Shinji Kamimura

    Physiology and Behavior   65 ( 4-5 )   665 - 669   1998

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    The correlation between social status and sperm motility of mice was investigated. From 5 to 15 weeks of age, mice were kept under two housing conditions, i.e., in pairs or in isolation. The social dominance in the paired mice was determined with the resident-intruder tests, which were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of reproductive organs, and serum testosterone were determined. It was revealed that the sperm motility of dominant mice was significantly higher than that of the subordinates. The sperm motility of the isolated mice was also significantly higher than the subordinates. It was suggested that the subordinate social status lowered sperm motility. (C) 1999 Elsevier Science Inc.

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  • Lowered sperm motility in subordinate social status of mice

    Koyama, S., Kamimura, S.

    Physiology and Behavior   65 ( 4-5 )   665 - 669   1998

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    The correlation between social status and sperm motility of mice was investigated. From 5 to 15 weeks of age, mice were kept under two housing conditions, i.e., in pairs or in isolation. The social dominance in the paired mice was determined with the resident-intruder tests, which were carried out from 8 to 15 weeks of age. At the end of 15 weeks of age, sperm activity, weights of reproductive organs, and serum testosterone were determined. It was revealed that the sperm motility of dominant mice was significantly higher than that of the subordinates. The sperm motility of the isolated mice was also significantly higher than the subordinates. It was suggested that the subordinate social status lowered sperm motility. (C) 1999 Elsevier Science Inc.

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  • 光学顕微鏡で計測する 細胞のミクロ探検 -見えないものを見る-

    上村慎治

    57 - 72   1997.4

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    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (other)   Publisher:日学選書B 日本学術会議事務局・日本動物学会関東支部編集、財団法人 日本学術協力財団発行  

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  • 高分解能顕微鏡で捉えた分子モーターの揺らぎ

    上村慎治

    生体分子モーターのしくみ シリーズ ・ ニューバイオフィジックス   144 - 157   1997.4

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  • 光学顕微鏡(1)-光学顕微鏡の基礎と位相差顕微鏡

    上村慎治

    5 ( 2 )   15 - 22   1997.4

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  • 高解像度運動解析

    上村慎治

    細胞   27 ( 8 )   1994.4

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  • Nanometer scale vibration in mutant axonemes of Chlamydomonas

    Yagi, T., Kamimura, S., Kamiya, R.

    Cell Motility and the Cytoskeleton   29 ( 2 )   177 - 185   1994

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    Flagellar axonemes of sea urchin sperm display high frequency (200-400 Hz) vibration with nanometer scale amplitudes in the presence of ATP [Kamimura and Kamiya, 1992: J. Cell Biol. 116:1443 -1454]. To investigate how various axonemal components affect the vibration, we examined vibration in wild-type and mutant axonemes of Chlamydomonas. At 1 mM ATP, wild-type axonemes underwent vibration at 100-650 Hz with amplitudes of 4-40 nm. This vibration was similar to, but less regular than, that in sea urchin sperm. Axonemes of the mutants ida1 and ida4 lacking part of the inner arm dynein underwent vibrations indistinguishable from that of wild-type. The mutant oda 1 lacking the entire outer arm underwent vibration at about half the wild-type frequency. Unexpectedly, the paralyzed mutants pf18 lacking the central pair and pf14 lacking the radial spokes displayed vibration with significantly higher frequencies and smaller amplitudes than those in the wild-type vibration. These results indicate that the high-frequency vibration is common to many kinds of mutant axonemes that lack various axonemal substructures, but that its manner is sensitive to the presence of outer arm dynein and the central pair/radial spoke system. Simultaneous measurements of amplitude and frequency in wild-type and mutant axonemes suggest that the velocity of microtubule sliding in vibrating axonemes is lower than the velocity of sliding under load-free conditions. The velocity is particularly low in pf18. A possible mechanism is proposed to explain the lower sliding velocity and vibration amplitude in the pf18 axoneme, based on an assumption that central pair/radial spoke system may work to regulate the switching of two antagonizing forces within the axoneme. (C) 1994 Wiley-Liss, Inc.

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  • Nanometer scale vibration in mutant axonemes of Chlamydomonas

    Toshiki Yagi, Shinji Kamimura, Ritsu Kamiya

    Cell Motility and the Cytoskeleton   29 ( 2 )   177 - 185   1994

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    Flagellar axonemes of sea urchin sperm display high frequency (200-400 Hz) vibration with nanometer scale amplitudes in the presence of ATP [Kamimura and Kamiya, 1992: J. Cell Biol. 116:1443 -1454]. To investigate how various axonemal components affect the vibration, we examined vibration in wild-type and mutant axonemes of Chlamydomonas. At 1 mM ATP, wild-type axonemes underwent vibration at 100-650 Hz with amplitudes of 4-40 nm. This vibration was similar to, but less regular than, that in sea urchin sperm. Axonemes of the mutants ida1 and ida4 lacking part of the inner arm dynein underwent vibrations indistinguishable from that of wild-type. The mutant oda 1 lacking the entire outer arm underwent vibration at about half the wild-type frequency. Unexpectedly, the paralyzed mutants pf18 lacking the central pair and pf14 lacking the radial spokes displayed vibration with significantly higher frequencies and smaller amplitudes than those in the wild-type vibration. These results indicate that the high-frequency vibration is common to many kinds of mutant axonemes that lack various axonemal substructures, but that its manner is sensitive to the presence of outer arm dynein and the central pair/radial spoke system. Simultaneous measurements of amplitude and frequency in wild-type and mutant axonemes suggest that the velocity of microtubule sliding in vibrating axonemes is lower than the velocity of sliding under load-free conditions. The velocity is particularly low in pf18. A possible mechanism is proposed to explain the lower sliding velocity and vibration amplitude in the pf18 axoneme, based on an assumption that central pair/radial spoke system may work to regulate the switching of two antagonizing forces within the axoneme. (C) 1994 Wiley-Liss, Inc.

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  • 運動をナノメートル精度で測る -顕微計測-

    上村慎治

    実験生物物理(石渡信一編,石川他著)   31 - 44   1993.4

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  • Tubulin protofilaments and kinesin-dependent motility. Reviewed

    Kamimura, S, Mandelkow, E

    J. Cell Biol.   ( 116 )   1443 - 1454   1992.8

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    DOI: 10.1083/jcb.118.4.865

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  • Tubulin protofilaments and kinesin-dependent motility

    S. Kamimura, E. Mandelkow

    Journal of Cell Biology   118 ( 4 )   865 - 875   1992.8

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    Microtubules are built of tubulin subunits assembled into hollow cylinders which consist of parallel protofilaments. Thus, motor molecules interacting with a microtubule could do so either with one or several tubulin subunits. This makes it difficult to determine the structural requirements for the interaction. One way to approach the problem is to alter the surface lattice. This can be done in several ways. Protofilaments can be exposed on their inside (C-tubules or "sheets"), they can be made antiparallel (zinc sheets), or they can be rolled up (duplex tubules). We have exploited this polymorphism to study how the motor protein kinesin attached to a glass surface interacts and moves the various tubulin assemblies.
    Microtubules glide over the surface along straight paths and with uniform velocities. In the case of C-tubules, approximately 40% glide similarly to microtubules, but a major fraction do not glide at all. This indicates (a) that a full cylindrical closure is not necessary for movement, and (b) that the inside surface of microtubules does not support gliding. With zinc sheets, up to 70% of the polymers move, but the movement is discontinuous, has a reduced speed, and follows along a curved path. Since zinc sheets have protofilaments alternating in orientation and polarity, this result suggests that in principle a single protofilament can produce movement, even when its neighbors cannot. Duplex microtubules do not move because they are covered with protofilaments coiled inside out, thus preventing the interaction with kinesin. The data can be explained by assuming that the outside of one protofilament represents the minimal track for kinesin, but smooth gliding requires several parallel protofilaments. Finally, we followed the motion of kinesin-coated microbeads on sea-urchin sperm flagella, from the flagellar outer doublet microtubules to the singlet microtubule tips extending from the A-tubules. No change in behavior was detected during the transition. This indicates that even if these microtubules differ in surface lattice, this does not affect the motility.

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  • High-frequency vibration in flagellar axonemes with amplitudes reflecting the size of tubulin

    S. Kamimura, R. Kamiya

    Journal of Cell Biology   116 ( 6 )   1443 - 1454   1992.3

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    Flagellar axonemes of sea urchin sperm display high-frequency (approximately 300 Hz) vibration with nanometer-scale amplitudes in the presence of ATP (Kamimura, S., and R. Kamiya. 1989. Nature (Lond.). 340:476-478). The vibration appears to represent normal mechanochemical interaction between dynein and microtubules because the dependence of the frequency on MgATP concentration is similar to that of the axonemal motility, and because it is inhibited by micromolar concentrations of vanadate. In this study a two-dimensional photo-sensor was used to characterize this phenomenon in detail. Several new features were revealed. First, the vibration was found to be due to a back-and-forth movement of the doublet microtubules along the axonemal length. Two beads attached to different parts of the same axoneme vibrated in unison, i.e., synchronized exactly in phase. This suggested that the outer doublet can be regarded as a stiff rod in vibrating axonemes. Second, evidence was obtained that the amplitude of the vibration reflected the number of active dynein arms. Third, under certain conditions, the vibration amplitude took stepwise values of 8 X N + 4 nm (N = 0, 1, 2, 3, or 4), indicating that the amplitude of microtubule sliding was limited by the size of tubulin dimer (8 nm) or monomer (4 nm). To explain this phenomenon, a model is presented based on an assumption that the force production by dynein is turned off when dynein is subjected to tensile force; i.e., dynein is assumed to be equipped with a feedback mechanism necessary for oscillation.

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  • High-frequency vibration in flagellar axonemes with amplitudes reflecting the size of tubulin

    Kamimura, S., Kamiya, R.

    Journal of Cell Biology   116 ( 6 )   1443 - 1454   1992

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    Flagellar axonemes of sea urchin sperm display high-frequency (~300 Hz) vibration with nanometer-scale amplitudes in the presence of ATP (Kamimura, S., and R. Kamiya. 1989. Nature (Lond.). 340: 476-478). The vibration appears to represent normal mechanochemical interaction between dynein and microtubules because the dependence of the frequency on MgATP concentration is similar to that of the axonemal motility, and because it is inhibited by micromolar concentrations of vanadate. In this study a two-dimensional photo- sensor was used to characterize this phenomenon in detail. Several new features were revealed. First, the vibration was found to be due to a back- and-forth movement of the doublet microtubules along the axonemal length. Two beads attached to different parts of the same axoneme vibrated in unison, i.e., synchronized exactly in phase. This suggested that the outer doublet can be regarded as a stiff rod in vibrating axonemes. Second, evidence was obtained that the amplitude of the vibration reflected the number of active dynein arms. Third, under certain conditions, the vibration amplitude took stepwise values of 8 x N + 4 nm (N = 0, 1, 2, 3, or 4), indicating that the amplitude of microtubule sliding was limited by the size of tubulin dimer (8 nm) or monomer (4 nm). To explain this phenomenon, a model is presented based on an assumption that the force production by dynein is turned off when dynein is subjected to tensile force
    i.e., dynein is assumed to be equipped with a feedback mechanism necessary for oscillation.

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  • 微小管

    上村慎治

    バイオメカニクスシリーズ,細胞のバイオメカニクス   143 - 177   1990.4

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  • 光学顕微鏡によるnm精度での運動の解析

    上村慎治

    30 ( 4 )   39 - 41   1990.4

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  • High-frequency, nanometre-scale vibration in 'quiescent' flagellar axonemes. Reviewed

    Kamimura, S, Kamiya, R

    Nature   340 ( 340 )   476 - 478   1989.8

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    DOI: 10.1038/340476a0

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  • High-frequency nanometre-scale vibration in 'quiescent' flagellar axonemes

    Shinji Kamimura, Ritsu Kamiya

    Nature   340 ( 6233 )   476 - 478   1989.8

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    THE movement of cilia and flagella is based on the interaction between dynein arms and microtubules coupled with ATP hydrolysis. Although it is established that dynein arms cause adjacent microtubules to slide, little is known about the elementary process underlying the force production. To look more closely at the mechano-chemical conversion mechanism, we recently developed an optical method for measuring a nanometre-scale displacement with a time-reslution better than 1 ms. We now report the detection of high frequency ( ∼ 300 Hz) vibration of sub-nanometre ampli-tude in non-beating flagellar axonemes. This vibration could reflect the movement of individual activated dynein arms.

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  • 分子間力を直接測定する新しい方法

    上村慎治

    38 ( 8 )   632 - 633   1989.4

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  • 光学顕微鏡で細胞運動を高精度で見る

    上村慎治

    4 ( 5 )   56 - 58   1989.4

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  • 細胞運動測定法 nm精度での顆粒運動の解析

    上村慎治

    39 ( 5 )   493 - 495   1988.4

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    DOI: 10.11477/mf.2425905198

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  • Direct measurement of nanometric displacement under an optical microscope

    Shinji Kamimura

    Applied Optics   26 ( 16 )   3425 - 3427   1987.8

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    A novel method has been developed to measure nanometric displacement under a conventional optical microscope. The magnified image of a pinhole was divided into two parts using a prism-shaped mirror. The difference of light intensity between the divided images was determined, which was proportional to displacement of the pinhole. Using a 5-Mm diam pinhole, the accuracy to determine displacement was -1 nm. Instead of a pinhole, polystyrene microbeads were used in the new method. Displacement of the microbeads was also measured with nanometric accuracy. This technique could be used to probe nanometric phenomena using optical microscopes. © 1987 Optical Society of America.

    DOI: 10.1364/AO.26.003425

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  • DIRECT MEASUREMENT OF NANOMETRIC DISPLACEMENT UNDER AN OPTICAL MICROSCOPE

    S KAMIMURA

    APPLIED OPTICS   26 ( 16 )   3425 - 3427   1987.8

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  • 新たな細胞運動系、キネシン・チュ-ブリン系の発見

    上村慎治

    27 ( 4 )   52 - 54   1987.4

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    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (other)   Publisher:生物物理  

    DOI: 10.2142/biophys.27.a180

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    Other Link: https://jlc.jst.go.jp/DN/JALC/00364705832?from=CiNii

  • Turbidimetric studies on microtubule sliding using the stopped-flow-light-scattering method

    Kamimura, S., Nakanishi, M., Yano, M., Shimizu, H.

    Experimental Cell Research   163 ( 1 )   186 - 190   1986.3

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    DOI: 10.1016/0014-4827(86)90571-9

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  • Turbidimetric studies on microtubule sliding using the stopped-flow-light-scattering method

    Shinji Kamimura, Mamoru Nakanishi, Masafumi Yano, Hiroshi Shimizu

    Experimental Cell Research   163 ( 1 )   186 - 190   1986.3

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    The turbidity of axonemes during active sliding of microtubules was analysed using the stopped-flow-light-scattering method with high time resolution. Flagella of sea-urchin spermatozoa were demembranated and used after a brief treatment with trypsin. The turbidity of the suspension of flagellar axonemes during ATP-induced disintegration was measured and its time course fitted to a single exponential function which yielded the rate of disintegration, R(l/sec). R coincided well with the velocity of microtubule sliding, V(μfx186-1 sec) as determined by cinematomicrographic analysis [13], i.e., R = 0.22 × V, r = 0.9973. It indicates that turbidimetry is a useful method with which to learn the sliding velocity of microtubules. From the dependency of R on temperature, Q10 of the sliding velocity was estimated to be 2.0-2.3 at 43-820 μM of MgATP. © 1986.

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  • ATP hydrolysis coupled to microtubule sliding in sea-urchin sperm flagella

    Shinji Kamimura, Masafumi Yano, Hiroshi Shimizu

    Journal of Biochemistry   97 ( 5 )   1509 - 1515   1985.5

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    Using sea urchin (Hemicentrotus pulcherimus) sperm flagella, ATP hydrolysis coupled to sliding movement of microtubules was investigated. Flagellar axonemes were pretreated with trypsin and the microtubules induced to slide by addition of ATP (50-1,000 μM) at 0-20°C. Motion-dependent hydrolysis of ATP was observed immediately after the addition of ATP, the rate of which was higher than that of steady state hydrolysis in axonemes without trypsin-treatment, or after complete disintegration. The rate of hydrolysis of ATP divided by the sliding velocity of microtubules reflects the ATP consumption necessary per unit distance of microtubule sliding. This parameter varied according to the experimental conditions in that it increased when the ATP concentration or temperature was decreased. Our results suggest that there is not a strict stoichiometric relationship between ATP hydrolysis and sliding distance in the dynein-tubulin system, indicating that the mechanochemical coupling is different from that in beating axonemes. © 1985, by the Japanese Biochemical Society.

    DOI: 10.1093/oxfordjournals.jbchem.a135206

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  • ATP hydrolysis coupled to microtubule sliding in sea-urchin sperm flagella

    Kamimura, S., Yano, M., Shimizu, H.

    Journal of Biochemistry   97 ( 5 )   1509 - 1515   1985

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  • Dynamic aspects of microtubule sliding in sperm flagella

    K. Takahashi, S. Kamimura

    Journal of Submicroscopic Cytology   15 ( 1 )   1 - 3   1983

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  • Dynamic aspects of microtubule sliding in sperm flagella

    Takahashi, K., Kamimura, S.

    Journal of Submicroscopic Cytology   15 ( 1 )   1 - 3   1983

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  • MICROTUBULE SLIDING IN REACTIVATED FLAGELLA

    K TAKAHASHI, C SHINGYOJI, S KAMIMURA

    SYMPOSIA OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY   ( 35 )   159 - 177   1982

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  • Microtubule sliding in reactivated flagella.

    K. Takahashi, C. Shingyoji, S. Kamimura

    Symposia of the Society for Experimental Biology   35 ( 35 )   159 - 177   1982

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    Recent experimental studies of microtubule sliding in demembranated sea urchin sperm flagella are described. A local iontophoretic application of ATP to a Triton-extracted flagellum elicits a local bending response whose form is in exact conformity with the predictions of the sliding microtubule model. Cinematographic analysis of the microtubule sliding initiated by treating fragments of demembranated flagella with trypsin in the presence of ATP reveals that the speed of sliding is almost constant. This implies that the speed does not depend on the number of dynein arms participating in the generation of sliding force. The distribution of apparent sliding velocities indicates that there is no difference in sliding velocity among the doublets. The sliding velocity depends on MgATP concentration in a manner consistent with Michaelis-Menten kinetics. The sliding velocity of doublets in trypsin-treated axonemes is close to the maximum velocity of relative sliding taking place between adjacent doublets in beating flagella reactivated at the same MgATP concentration.

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  • Direct measurement of the force of microtubule sliding in flagella

    Shinji Kamimura, Keiichi Takahashi

    Nature   293 ( 5833 )   566 - 568   1981

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    The movement of eukaryotic cilia and flagella is caused by ATP-driven active sliding between the doublet microtubules1-3, and the force for the sliding is believed to be generated by the dynein arms of the A-tubule interacting with the B-tubule of the adjoining doublet. To understand the mechanochemical basis of this force-generating reaction and to correlate it with the overt motile behaviour of cilia or flagella, it is important to quantify the force exerted by the dynein arms. Attempts have been made to estimate this force from the bending moment generated by the whole organelle4,5, but the estimation was of necessity hypothetical because the mechanism by which sliding is coupled with bending is poorly understood. Microtubule sliding without bending can be induced in vitro if trypsin-2 or elastase 6-treated axonemes are perfused with a solution containing ATP. By attaching glass microneedles to such axonemes, we have now directly determined the sliding force at various ATP concentrations. © 1981 Nature Publishing Group.

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  • Direct measurement of the force of microtubule sliding in flagella

    Kamimura, S., Takahashi, K.

    Nature   293 ( 5833 )   566 - 568   1981

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    DOI: 10.1038/293566a0

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Presentations

  • Novel image processing technique for the analysis of life span and activity in Drosophila melanogaster

    R. Hyodo, R. Aoki, J. Ou, H. Katoh, S. Kamimura

    The 94th Annual Meeting of the Xoological Sociery of Japan  2023.9 

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  • Characteristics of green pigment derived from Clypeaster japonicus

    A.SHiroyama, T. Nishida, M. Kohno, H. Katoh, S. Kamimura

    The 94th Annual Meeting of the Xoological Sociery of Japan  2023.9 

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  • Properties of green pigments isolated from sea biscuits, Clypeaster japonicus.

    Tomoya Nishida, Mitsuko Kawano, Yusuke Kato, Shinji Kamimura

    The 93rd Annual Meeting of the Zoological Sociery of Japan 1C23-1630  ( Waseda University )   2022.9  the Zoological Sociery of Japan

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  • Green pigment-producing cells in Clypeaster japonicus embryo.

    Hiroto Amano, Mitsuko Kawano, Yusuke Kato, Shinji Kamimura

    The 93rd Annual Meeting of the Zoological Sociery of Japan IC24-1645  ( Waseda University )   2022.9  the Zoological Sociery of Japan

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  • Determination of longevity and activity magnitude by image processing inDrosophila melanogaster.

    Ryoma Aoki, Ritsuki Hyodo, Haruna Kato, Syousei Ou, Shinji Kamimura

    The 93rd Annual Meeting of the Zoological Sociery of Japan 3H36-1530  ( Waseda University )   2022.9  the Zoological Sociery of Japan

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  • Novel isolated ciliary dynein structure revealed by negative stain E

    Y. Lei, H. Imai, R. Yamamoto, R. Shimo, S. Kamimura, T. Yagi, N. Kajimura, M. Hirose, T. Kato, K. Mitsuoka, T. Kon

    第59回 日本生物物理学会年会  ( On Line )   2021.11  the Biophisical Society of Japan

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    Dyneins are phylogenetically classified into three subfamilies, ciliary, cytoplasmic and intraflagellar transport (IFT) dyneins. It is widely known that cytoplasmic and IFT dyneins adopt an inactivated conformation called the phi structure. In contrast, such a distinct inactivation state was not evident for ciliary dynein. Here we report that one of the ciliary dynein species has a novel structure that is very similar to the phi structure. Based on the structural analysis, we propose a novel shutdown mechanism common to all three subfamilies of dynein.

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  • Structural dynamics of native microtubules on cooling: anisotropic and hysteretic structural changes depending on temperature

    S. Kamimura, T. Yagi, Y. Kondo, J. Estévez-Gallego, D. Lucena-Agell, J. F. Díaz, H. Iwamoto

    The 59th Annual Meeting of the Biophysical Society of Japan  ( On Line )   2021.11  the Biophysical Society of Japan

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    Microtubules (MT) are involved in essential cellular functions. One known characteristic of them is rapid disassembly to tubulin subunits upon cooling, a process whose molecular basis is not yet understood. We hypothesize that the conformational changes in tubulin accumulate strain of MT that triggers disassembly. To test this hypothesis, we analyzed the X-ray fiber diffraction patterns of native MTs during rapid cooling from 37 to 10℃ (<20s). We found that shape changes of MT on cooling was anisotropic, i.e., the magnitude of shrinkage was different between longitudinal and diameter directions. Detailed time-course analysis showed that the shrinkage and expansion are hysteretic. The present study would help us to understand how MTs become instable at low temperatures.

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  • Bioconvection of the harmful algae Chattonella

    Mina Nakahara, Atsuto Kobayashi, Shinji Kamimura

    第43回エアロ・アクアバイオメカニズム学会講演会  ( ネット開催 )   2021.3  Society of Aero Aqua Bio-mechanisms

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    Chattonella is marine algae that causes HAB at Seto Inland Sea in Japan. When collected in a shallow petri dish, they start gradual accumulation in a few minutes and forms specific spotted or branching patterns with nonhomogeneous cell distributions. Since this phenomenon of bioconvection is expected to be tightly correlated with HAB mechanisms, we are executing the image analysis of accumulation behavior of Chattonella using a spatial statistical technique. Examples to show novel quantitative descriptions of bioconvection we found are shown.

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  • Variogram and correlogram assay of cell motility: Bioconvection in harmful algae Chattonella

    Mina Nakahara, Atsuto Kobayashi, Shinji Kamimura

    The Society of Biophysics of Japan  ( Online )   2020.9  The 58th Annual Meeting of the Biophysical Society of Japan

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    Chattonella is single-celled marine algae that cause harmful algal blooms (HAB). When collected in a shallow petri dish, they start gradual accumulation to form specific patterns with a non-uniform distribution. This phenomenon of bioconvection is considered to be closely related to the HAB formation mechanism in fields. In order to understand the mechanism of HAB development, we executed the image analysis of collective swimming behavior of Chattonella marina var. ovata (Raphidophyceae) using spatial statistical techniques as well as high-speed-video microscopy. Here, we present how this quantitative analysis, variogram or correlogram, an empirically tool in geostatics, can be useful to describe the pattern of bioconvection.

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  • English]:Dynamic changes of tubulin dimer configurations on a scale of sub-second revealed by high flux X-ray fiber diffracti Invited

    Shinji Kamimura

    he 57th Annual Meeting of the Biophysical Society of Japan  ( Seagaia Convention Center )   2019.9  Biophysical Society of Japan

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    Microtubules are assembled from tubulin dimers that are arranged in a semi-crystal lattice with high regularity. Dynamics of tubulin dimer within microtubules is thus expected to be directly affecting the stability of microtubules, however, it has been difficult to quantitatively describe such properties. By analyzing the time course of pattern changes in X-ray fiber diffractions from aligned microtubules with a high-flux synchrotron beam of BM40XL (SPring-8), we found microtubules showed rapid elongation in the axial tubulin repeat and the concomitant increase of structural flexibility with a time constant of about 0.2 s after applying paclitaxel, microtubule-stabilizer. Contrasting effects were found for laulimalide, another type of microtubules stabilizer.

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  • etermination of accurate axial tubulin periodicity in native microtubules under physiological conditions

    Shinji Kamimura

    The 90th Annual Meeting of Zoological Society of Japan  ( Osaka City University )   2019.9  Zoological Society of Japan

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  • 微小管内のチューブリン分子の安定性・可塑性・柔軟性を目安にした微細動態解析

    上村慎治, 今井洋, 八木俊樹, 岩本裕之

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019.3  日本動物学会関東支部

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    微小管内にはチューブリン分子が、結晶のように整然とラセン状格子配置している。これまで抗がん剤として、種々の微小管安定化剤が開発されて来ているが、微小管構造の安定性とチューブリンの分子構造変化とを関連づけて調べる良い方法がなかった。我々はX線繊維回折法を用いて、タキソールとラウリマライドの2種の安定化剤の効果の明確な違いを明らかにできたので報告する。両者ともに秒単位で微小管構造の変化を引き起こすことがわかったが、チューブリン分子の縦・横方向の相互作用に関して異なる効果を示すことも明らかとなった。

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  • ラフィド藻シャトネラの遊泳停止と赤潮発生機構

    中原 美奈, 成田 大樹, 和田 祐子, 鈴木 雄大, 田中 昂輝, 今井 洋, 上村慎治

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019.3  日本動物学会関東支部

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    シャトネラ(Chattonella marina var. obata)は、栄養塩類や海水温の条件が整うと急速に増殖・集積する赤潮の原因となるラフィド藻である。小型シャーレ内に培養した高密度のシャトネラを静置すると、数分内に自発的な集積を開始し、時間経過と共に集団で移動する生物対流現象を示すことがわかった。これが実験室内で再現される赤潮現象であると考えられる。この現象は細胞の密度を上げることで促進され、海水中のCa2+濃度を減らすことで抑制されることがわかった。位相差顕微鏡を使ったシャトネラの遊泳行動観察から、細胞密度を上昇させると互いにぶつかり合う頻度が増加し、その機械的な刺激によって短時間の鞭毛打停止反応が起こり、同時に数秒間遊泳停止することがわかった。この鞭毛打停止反応は海水中のCa2+濃度を減らすことで抑制されることもわかった。ここで発見された鞭毛停止反応が、赤潮における細胞集積反応の1つの重要な要因になっていると考えられる。

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  • タコノマクラの作る緑色色素の分泌機構と役割

    加藤佑亮, 中村洋輝, 鬼頭玲賀, 河野美都子, 中原美奈, 上村慎治

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019.3  日本動物学会関東支部

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    タコノマクラ(Clypeaster japonicus)は、KCl神経刺激や殻部損傷によって緑色の色素を体外へ分泌するため、この色素は生体防御機構と深く関わっていると考えられる。その色素の特性は、Goodwin & Fox(1955)の吸収スペクトルに関する報告しかない。本研究で緑色色素の特性を詳細に調べた結果、親水性の高い高分子量の成分であることがわかり、他種のウニから採取されるナフトキノン系の赤色色素であるechinochromeやspinochrome等とは大きく異なる点、さらに、スカシカシパンにも共通する特性の色素があることがわかった。4腕幼生でもすでに生成機能が備わっており、腕部の機械的な刺激により内胚葉組織付近で分泌される点、その分泌にAchが関わっていることもわかった。

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  • キンギョ精子の運動活性化について

    加木下原自, 奥野誠, 上村慎治

    (社)日本動物学会第71回関東支部大会  ( 中央大学理工学部 )   2019.3  日本動物学会関東支部

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    魚類の精子は、生殖器内と異なる環境に暴露されことで運動が誘発される。今回、キンギョの精子ではイオンや糖の種類にかかわらず、240 mOsmより高い浸透圧で運動が停止することがわかり、運動停止に関しては、K+濃度よりも、高浸透圧が重要であることが示唆された。また、精子の細胞膜を除去し再活性化を試みたところ、ATPが存在すればCaCl2やcAMPを添加していない溶液にても精子の再活性化がみられ、Ca2+、cAMPともに、精子運動開始の細胞内シグナル伝達において必須でない可能性が示唆された。

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  • Biochemical and structural characterization of taxoid microtubule-stabilizing agents

    Estevez-Gallego J, Kamimura S, Balaguer-Perez, F, Lucena-Agell D, Diaz J-F

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018.5 

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  • Structural basis for the GTP activation of tubulin

    Dlaz, Jose-Fernando

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018.5 

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  • Dynamic changes in microtubule structure observed on a time scale of seconds by X-ray fiber diffraction

    Kamimura S, Imai H, Yagi T, Iwamoto H

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018.5 

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    Microtubules are key components of the cytoskeleton in eukaryotic cells. Their dynamic conversions between assembled microtubules and free tubulin dimers in cytoplasm are precisely controlled along with the states of cell activities. One of the most fundamental questions of us is how the microtubule dynamics and its structural properties are related to the chemical states of tubulin dimers during GTP-hydrolysis, with the existence of other MAPs and tubulin-binding chemicals.|rn||rn| X-ray fiber diffraction is one of the most powerful techniques to collect the structural information of native microtubules in physiological solutions without fixation, crystallization or freezing. In the present study, we examined the structural changes of microtubules on a time scale of seconds by combining our original technique for shear-flow alignment of microtubules [Sugiyama et al, 2009; Oiwa et al., 2009; Kamimura et al., 2016] with a high-flux beam line at SPring-8 (BL40XU). Our observations clearly showed that binding with paclitaxel induced the decrease of mean microtubule diameter as well as the increase of axial tubulin repeat within a second, indicating configuration changes of tubulin di

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  • Conformational switching of microtubule and cooperative binding of kinesin-1 for polarized transport

    Shima Tomohiro

    EMBO Symposium: Microtubules from Atom to Complex Systems  2018.5 

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  • Biochemical and structural characterization of taxoid microtubule-stabilizing agents

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018 

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  • Structural basis for the GTP activation of tubulin

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018 

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  • Dynamic changes in microtubule structure observed on a time scale of seconds by X-ray fiber diffraction

    EMBL Symposium: Microtubules: From Atoms to Complex Systems  2018 

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  • Conformational switching of microtubule and cooperative binding of kinesin-1 for polarized transport

    EMBO Symposium: Microtubules from Atom to Complex Systems  2018 

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  • Conformational switching of microtubule and cooperative binding of kinesin‐1 as a base for polarized transport (B115)

    Shima T, Morikawa M, Kaneshiro J, Kambara T, Kamimura S, Yagi T, Iwamoto H, Uemura S, Shigematsu H, Ichimura T, Watanabe TM, Nitta R, Okada Y, Hirokawa N

    ASCB-EMBO Meeting 2017  2017.12 

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    Kinesin-1, the founding member of kinesin superfamily proteins, is known to use only a subset of microtubules for transport in living cells. This biased use of microtubules is proposed as the guidance cue for polarized transport in neurons, but the underlying mechanisms are still poorly understood. Here we report that there is a positive feedback in the binding of kinesin-1 to the GDP-microtubule, which spontaneously produces high affinity microtubules from other low-affinity microtubules. This high affinity state requires the binding of kinesin-1 in the nucleotide-free state. Microtubules return to the initial low affinity state by washing out the binding kinesin-1 or by the binding of AMPPNP to kinesin-1. X-ray fiber diffraction, fluorescence speckle microscopy and second harmonic generation microscopy, as well as cryo-EM, collectively demonstrated that the binding of nucleotide-free kinesin to GDP-microtubule changes the conformation of GDP-microtubule to a conformation close to the GMPCPP-microtubule.

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  • “Double wave” of Octopus sperm flagella under high viscous condition

    Yuuko Wada, Yoshihiro Mogami, Shinji Kamimura

    Annual Meeting of Zoological Society of Japan  2017.9 

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    マダコは体内受精と体外受精の中間的な生殖戦略を取る。交尾は行わず、オスは精莢と呼ばれるカプセル入りの精子をメスに手渡し、精子はメスの体表上で精莢から放出され、その後、貯精嚢へと移動する。メスの貯精嚢内で精子は数週間保存され適宜使われる。単離した精莢から海水中に放出された精子を通常海水中で観察すると、頭部を折り曲げその場で回転する運動と、頭部を振って前進する運動とを繰り返しながら絡み合って束を形成する。マダコ精子は約330 μmの長い鞭毛をもち、通常海水中で鞭毛の屈曲は根元と先端では顕著だが、根元から100-200 μmの中間部分ではあまり見られない。海水中に分子モーターダイニンの阻害剤であるシリオブレビン(文献1)を加えると、シリオブレビンAでは波形変化は見られなかったがシリオブレビンDでは後半部分が極端に大きく屈曲した(日本動物学会第86回新潟大会)。|rn| 2%メチルセルロース(1500 cPs)を加えて粘度を上げた海水中で

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  • プラナリアにおける腹部繊毛の形態観察

    Zoological Society of Japan  2017.9 

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  • “Double wave” of Octopus sperm flagella under high viscous condition

    Annual Meeting of Zoological Society of Japan  2017 

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  • Conformational switching of microtubule and cooperative binding of kinesin‐1 as a base for polarized transport (B115)

    ASCB-EMBO Meeting 2017  2017 

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  • Flagellar wave form of Octopus vulgaris sperm in high viscosity medium

    Yuuko Wada, Shinji Kamimura

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016.11 

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    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan|rn||rn||rn||rn|Reproduction strategy of octopus is just the intermediate between internal and external fertilization. During mating, male octopus deliver pieces of spermatophore to females. Spermatozoa, after being released from the spermatophores placed on the body surface of females, move or swim towards the spermathicae, sperm storage organs in female oviducts. In spite of detailed descriptions so far on such unique behaviors of Octopus mating, there has been few reports investigating the sperm physiology of Octopus vulgaris. Their flagellar length was about 330 μm with a head of about 25 μm long. We found the manner of flagellar bend propagation was different from usual flagellar motions. It seemed to be propagating along the entire length of flagellar shaft in a non-continuous manner, i.e., smooth bends propagated along the proximal region of ca. 100 μm and then diminished at a middle part of the flagellum (100-200 μm from the base) where flagellum look almost straight, then flagellar bends reappeared around 200 μm from the base. We also found there was difference in the wave

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  • Effects of high pressure on the motility of sea urchin sperm flagella

    Hiroshi Imai, Masayoshi Nishiyama, Yoshie Harada, Shinji Kamimura

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016.11 

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    Sea urchin spermatozoa released from males swim towards eggs in seawater for fertilization in the intertidal zone where sea urchins live. There are many other species living in the deep sea, or some of them accidentally may fall down to the deep sea by gravity after storm. Thus, the fertilization could be occurring under the stress of high or various different pressures. However, there are no report describing how the sperm swimming and flagellar motility is affected under such high pressure conditions.|rn|In order to understand the effects of pressure on motility of sea urchin spermatozoa, we examined sperm motility of sea urchin, Anthocidaris crassispina, under high pressure of seawater in the presence or in the absence of 10 mM CaCl2. Without Ca2+, consistently with previous studies, we observed the circular swimming path of sea urchin sperm on the glass surface under the condition of atmospheric pressure. The path of circular swimming was similar to that of spermatozoa observed in the seawater containing 10 mM CaCl2. We interpreted that the pumping activity of Ca2+ channels or other transporting systems (e.g. Na+/Ca2+ exchanger) can maintain the intracellular Ca2+ concentration

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  • Regulation of cilia and flagella bending movements through the change of axoneme diameter

    Toshiki Yagi, Shinji Kamimura, Hiroyuki Iwamoto

    The 54th Annual Meeting of the Biophysical Society of Japan  2016.11 

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  • Direct flow analysis around the beating flagella of sea-urchin sperm cells supporting the slender body theory of micro-swimmers

    Miyashiro D, Wada Y, Shihira-Ishikawa I, Miya-waki A, Kamimura S

    31st International Congress on High-speed Imaging and Photonics  2016.11 

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    For the efficient chemotaxis and fertilization, it should be a crucial problem for animal spermatozoa to swim in the medium without disturbing the concentration gradient of chemical attractants derived from unfertilized eggs. Precise analysis and description of micro-flows occurring around swimming sperm cells was expected to help us to understand the chemotaxis phenomena more in details, however, there were many technical prob-lems to be solved. In the present study, using an in-situ storage image sensor (ISIS) equipped on a differential interference contract (DIC) microscope with a high-intensity xenon-lamp illumination, we successfully record-ed clear images of swimming sea-urchin sperm cells at 6,000 fps in a medium containing micro-particles with 0.1 μm diameter to directly visualize medium flows around bending flagella. We processed the acquired DIC images and traced precise motions of sperm flagella as well as moving free micro-particles with a 0.3-ms time-resolution.|rn|By comparing with theoretical flows expected from the slender body theory (SBT), we concluded that ob-served flows around micro-swimmers such as beating flagella under microscopes are classified as stokes f

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  • Swimming motility of deep-sea bacteria measured by high-pressure microscopy

    Masayoshi Nishiyama, Chiaki Kato, Hiroshi Imai, Shinji Kamimura, Yoshie Harada

    The 54th Annual Meeting of the Biophysical Society of Japan  2016.11 

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  • Determination of accurate axial tubulin repeat in GDP-microtubules.

    Kamimura S, Imai H, Yagi T, Shima T, Okada Y, Iwamoto H

    54th Meeting of the Biophysical Society of Japan  2016.11 

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    The axial repeat of tubulin molecules in microtubules is roughly 4 nm and has conventionally been used as one of the most convenient standard scales to estimate molecular or organelle size for electron microscopy. However, in our previous report, the axial repeat varied depending on experimental conditions, e.g., temperature, with microtubule stabilizer and GTP-hydrolysis states. It was also shown that the tubulin axial repeat in porcine brain GTP-microtubules, which are mainly composed of GDP-tubulin, was almost constant with a low coefficient of thermal expansion of 3.7×10-5/degree. Here, we determined the accurate repeat of GDP-tubulin molecules after careful revision of camera length, X-ray wavelength as well as the angles of specimen, beam and detector tilting.

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  • New features of the structural dynamics of native microtubules revealed X-ray fiber diffraction

    22nd International Congress of Zoology (ICZ) & the 87th meeting of the Zoological Society of Japan  2016.11 

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    X線繊維回折法には、生体試料中の周期構造を直接反映したの回折信号を、< 0.1nmもの高精度でリアルタイム解析できる点で、他手法にはない優れた利点がある。本研究では、一般に重合・脱重合の平衡状態を繰り返すために、動的な構造ゆらぎも大きいと想像される微小管を使った解析結果を紹介する。GTP加水分解や微小管安定化剤添加による構造変化が見られることがわかったが、通常のポリマーには見られない特性、負の熱膨張特性が観察されることがわかった。細胞骨格の示すのユニークな特性について議論したい。

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  • A novel function of kinesin-I: changing microtubule conformation that accelerates successive kinesin binding.

    Tomohiro Shima, Manatsu Morikawa, Junichi Kaneshiro, Taketoshi Kambara, Shinji Kamimura, Toshiki Yagi, Hiroyuki Iwamoto, Taro Ichimura, Tomonobu Watanabe, Sotaro Uemura, Ryo Nitta, Yasushi Okada, Nobutaka Hirokawa

    The 54th Annual Meeting of the Biophysical Society of Japan  2016.10 

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  • X-ray fiber diffraction revealing the dynamic changes of native microtubule structure

    大阪大学蛋白質研究所セミナー 第6回分子モーター討論会「分子モーター研究の最前線」  2016.7 

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  • X-ray fiber diffraction analysis of microtubule revealing the dynamic changes of axial tubulin repeats in native microtubules

    S. Kamimura, Y. Fujita, Y. Wada, T. Yagi, T. Shima, Y. Okada, H. Iwamoto

    2016 EMBL Symposia Microtubules, Microtubules: From Atoms to Complex Systems  2016.5 

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    One of the most fundamental questions on the molecular mechanisms of microtubule assembly-disassembly dynamics is how the structural stability of microtubules is correlated with the variation of molecular conformation of tubulin dimers within microtubules. To address this issue, we applied our technique for the rapid shear-flow alignment of biological filaments in a physiological buffer medium, enabling us to acquire the structural periodicity data from native microtubules by X-ray fiber diffraction under solution conditions. We could classified microtubules into three main groups on the basis of distinct mean axial tubulin repeats and microtubule diameters that varied depending on GTP hydrolysis and the content of paclitaxel. The paclitaxel effects to induce the elongation of the mean axial repeat of tubulin was observed to be completed within 30 s. It was also suggested that this elongation effects were appearing in a cooperative manner from the observation of microtubules with subthreshold paclitaxel content. Another interesting feature we found was the variation in the temperature dependency of axial tubulin repeat in GTP-, GMPCPP- and GTPγS-microtubules with/without paclitaxe

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  • X線繊維回折で探る微小管構造の動態

    大阪大学蛋白質研究所セミナー 第6回分子モーター討論会「分子モーター研究の最前線」  2016 

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  • キネシンによる微小管の構造変化

    日本生物物理学会 第54回年会  2016 

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  • 高圧力顕微鏡法による深海微生物の遊泳運動観察

    日本生物物理学会 第54回年会  2016 

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  • 微小管内GDP-チューブリンの精密な周期決定

    第54回 日本生物物理学会  2016 

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  • X線繊維回折で解き明かす微小管構造の動態

    第87回 日本動物学会(第22回 国際動物学会)  2016 

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  • X-ray fiber diffraction analysis of microtubule revealing the dynamic changes of axial tubulin repeats in native microtubules

    2016 EMBL Symposia Microtubules, Microtubules: From Atoms to Complex Systems  2016 

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  • Flagellar wave form of Octopus vulgaris sperm in high viscosity medium

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016 

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  • Effects of high pressure on the motility of sea urchin sperm flagella

    The 22nd International Congress of Zoology & the 87th meeting of the Zoological Society of Japan  2016 

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  • Regulation of cilia and flagella bending movements through the change of axoneme diameter

    The 54th Annual Meeting of the Biophysical Society of Japan  2016 

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  • Direct flow analysis around the beating flagella of sea-urchin sperm cells supporting the slender body theory of micro-swimmers

    31st International Congress on High-speed Imaging and Photonics  2016 

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  • Microtubule dynamics revealed by the X-ray fiber diffraction analysis

    Shinji Kamimura, Hiroyuki Iwamoto

    Biophysical Society of Japan. #53 Annual meeting  2015.9 

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    Microtubules play a variety of functions in cells. Recent studies using high-resolution cry-electron micrography and X-ray fiber diffraction revealed that multiple conformational states of tubulin give rise to structural polymorphism in microtubule lattice, which may be crucial for diverse physiological functions of microtubules. Multiple conformations of tubulin also underlie dynamic instability of microtubules, where the balance between assembly and disassembly is stochastically switched in a nucleotide-dependent manner. In this session, we introduce the forefront in the field of microtubule, aiming to share new findings and insights about the conformational dynamics of tubulin and microtubule.

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  • X-ray fiber diffraction analysis revealed a highly flexible state of GTPγS-tubulin in the microtubules

    Shinji Kamimura, Yosuke Fujita, Yuuko Wada, Tomohiro Shima, Yasushi Okada, Toshiki Yagi, Hiroyuki Iwamoto

    #53 Annual Meeting, Biophysical Society of Japan (Kanazawa)  2015.9 

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    Our major question is how tubulin states are related to the stability of microtubules. To address the question, we used our novel technique for the quick shear-flow alignment of biological filaments, which enabled us to acquire fine X-ray fiber diffraction signals from native microtubules in a few seconds. We found that microtubules could be classified into three major groups with distinct axial periodicities of tubulin, which varied depending on the temperature of solution, the state of GTP-hydrolysis and the contents of microtubule stabilizers. It was also revealed that the GTPγS-tubulin showed the widest variation in the longitudinal tubulin periodicity in situ, suggesting a highly flexible state of GTP-tubulin in the microtubules.

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  • 柔軟性に富んだ微小管内GTPγS-チューブリン分子

    日本生物物理学会 第53回年会(金沢)  2015 

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  • Microtubule dynamics revealed by the X-ray fiber diffraction analysis

    Biophysical Society of Japan. #53 Annual meeting  2015 

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  • Length variations of tubulin molecules within native axonemal microtubules

    上村慎治, 岩本裕之

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014.11 

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    Tubulin is one of the major polypeptides that construct the axonemes of cilia/flagella. Thus, the mechanical or chemical properties of tubulin in axonemes would be optimized for the bending motion or against mechanical stresses given by external liquid. In the present study, with an X-ray fiber diffraction technique, the length changes or variations of tubulin dimer periodicity within microtubules (MT) was investigated with a high precision (~ 0.01 nm). We found there are two main states of tubulin periodicity within usual porcine brain MTs (intracellular singlet MTs), i.e., short and long configurations with a length change by about 3% depending on temperature and taxol binding (MT stabilizer). However, the tubulin repeat within sea-urchin sperm flagella showed just the intermediate of the two states of brain MTs. Here we postulate a new hypothesis, the mechanical capacity of tubulin molecules inside axonemes, which would serve a certain allowance of bending compression-decompression inside MTs.

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  • Coordinated activity of ciliary beats and muscle contractions is required for the gliding motion in Planarian

    Gentarou Sugita, Shinji Kamimura

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014.11 

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    The gliding motion of flatworms (Planarian, Platyhelminthes) is primarily generated by the effective strokes of ventral cilia. The maximum motion rate of ca. 3-5 mm/s appears to be one of the fastest rates reported for the ciliary motility. Our assumption is there would be fine optimization or regulation of the motile mechanisms to accomplish such a high gliding performance. In the present study, we used different types of specific motility blockers, blebbistatin/BDM for muscular myosin II and ciliobrevin for axonemal dyneins. Blebbistatin blocked peristaltic motions and depressed the half-cylindrical shapes of flatworm body (making flatworms more planar). BDM also blocked gliding motions. We also fund ciliobrevin completely inhibited the gliding motion in minutes, and induced peristaltic shape changes. From our observations, it was suggested that both muscle contraction and ciliary motion, some coordination between them, would be required for the efficient motion of gliding.

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  • X-ray diffraction study of aligned collagen fiber

    Yasunobu Sugimoto, Sakurako Hayashi, Sayaka Hayashi, Nobuhisa Watanabe, Shinji Kamimura, Takanori Kihara

    2014.9 

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  • Inhibition of ciliobrevin D on motility of sea urchin sperm flagella

    Wada Yuuko, Baba Shoji, Kamimura Shinji

    日本動物学会  2014.9 

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    ウニ精子にダイニン特異的な低分子阻害剤cliobrevin A(HPI4)を投与すると鞭毛打周波数が濃度依存的に低下し、高濃度では根元の非対称性が大きな運動となることを前回大会で報告した。今回、合成アナログのciliobrevin D をウニ精子に投与したときの阻害効果をciliobrevin A と比較した。周波数への効果はciliobrevin A と同様だが、波形への効果として鞭毛全長にわたり運動非対称性がさらに増し、鞭毛が頭部の周りに巻きつくものが多数みられた。ダイニン種の違いによる阻害効果の差という観点で考察を加える。

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  • Dynamic changes of the structure of double helix DNA in shear flow

    藤田洋介, 中澤亘將, 岩本裕之, 上村慎治

    日本生物物理学会  2014.9 

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  • Dynamic changes of the structure of double helix DNA in shear flow

    藤田洋介, 中澤亘將, 岩本裕之, 上村慎治

    日本生物物理学会  2014.9 

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  • Paclitaxel添加直後の微小管構造変化

    清原恵, 中澤亘將, 藤田洋介, 和田祐子, 八木俊樹, 岩本 裕之

    日本動物学会  2014.9 

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  • Molecular machinery regulating photomovement of Euglena

    Shimabukuro, SAP, ミドリムシにおける光運動制御マシナリーの解明|rn|Molecular machinery regulating photomovement of Euglena|rn, 岩崎 憲, 宮崎 直幸, 伊関 峰生, 長谷川 浩司, 成田 哲博

    日本生物物理学会  2014.9 

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  • ミドリムシにおける光運動制御マシナリーの解明

    日本生物物理学会  2014 

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  • プラナリア滑走運動における繊毛・筋運動協調関係

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014 

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  • X-ray diffraction study of aligned collagen fiber

    2014 

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  • Inhibition of ciliobrevin D on motility of sea urchin sperm flagella

    2014 

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  • Dynamic changes of the structure of double helix DNA in shear flow

    2014 

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  • Length variations of tubulin molecules within native axonemal microtubules

    6th International Symposium on Aero-aqua Bio-Mechanisms (ISABMEC2014)  2014 

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  • Dynamic changes in axial tubulin repeats in native microtubules revealed by X-ray fiber diffraction

    Dynein 2013 International Workshop (Kobe)  2013.10 

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  • Dynamic changes of the axial pitch of tubulin repeat in live microtubules revealed by x-ray fiber diffraction

    Shinji Kamimura, Yosuke Fujita, Yuuko Wada, Hiroyuki Iwamoto

    The 51st Annual Meeting of the Biophysical Society of Japan  2013.10 

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  • X ray fiber diffraction of microtubules: Tubulin pitch variations depending on medium temperature, GTP hydrolysis and taxol binding indicating dynamic

    Kamimura Shinji, Fujita Yosuke, Wada Yuuko, rn|Iwamoto Hiroyuki

    The 84th Annual Meeting of Zoological Society of Japan  2013.9 

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    溶液内でtubulin dimerは、curved/straight の2 状態をとると考えられている。微小管内でのtubulin の動的構造変化は、微小管の安定性に大きく影響すると考えられるが、まだ、その詳細を調べた研究報告はない。X 線微小管繊維回折の解析から、taxol 濃度、温度、GTP 加水分解に依存してtubulin ピッチが変わることがわかった。微小管内でtubulin dimer がshort/long の2 状態をとる可能性が考えられる。

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  • Effects of dynein specific inhibitor, ciliobrevin on the flagellar motility of sea urchin spermatozoa.

    Yuuko Wada, Shinji Kamimura

    The 84th Annual Meeting of the Zoological Society of Japan  2013.9 

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    細胞質ダイニンの特異的な阻害剤であるciliobrevin の軸糸ダイニンに対する効果を調べた。この物質は低分子で膜透過性を持つので、除|rn|膜していないウニ精子に直接投与した場合に効果を見ることが期待された。予想通り、ciliobrevin は濃度依存的にウニ精子の鞭毛打周波数を下げ、このとき波形の変化も観察された。つまり、内腕、外腕ダイニンの両方に効果があることが示された。

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  • ウニ精子鞭毛運動におけるciliobrevinの効果

    公益社団法人 日本動物学会 第84回大会  2013 

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  • 微小管X 線繊維回折:チューブリンピッチの動的変化

    日本製物物理学会第51回年会(京都)  2013 

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  • Xray fiber diffraction of microtubules: Tubulinpitch variations depending on medium temperature, GTPhydrolysis and taxol binding indicating dynamic

    The 84th Annual Meeting of Zoological Society of Japan  2013 

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  • Dynamic changes in axial tubulin repeats in native microtubules revealed by X-ray fiber diffraction

    Dynein 2013 International Workshop (Kobe)  2013 

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  • Direct imaging of the world of low Reynolds number hydrodynamics

    D. Miyashiro, Y. Wada, I. Shihira-Ishikawa, A. Miyawaki, S. Kamimura

    The Fifth International Symposium on Aero Aqua Bio-mechanisms, ISABMEC 2012 Taipei  2012.8 

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    To clarify the fluid dynamics of swimming microorganisms, a technique to visualize fluid flows under an optical microscope with appropriate spatial and temporal resolution is required. In the present study, we used sea-urchin spermatozoa that have long flagella with planar beat patterns and are suited for precise beat form analysis. Differential inter ference microscopy (DIC) images of the beating sperm flagella and particles included in the medium were recorded with a rate of 6,000 fps. As has been theoretically expected, spermatozoa were observed to swim in a stable medium without perturbation of surrounding fluids. This is the first direct evidence to show how fluid flows around whipping flagella under the condition of low Reynolds number.

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  • 微小管の非等方的冷却収縮

    上村慎治, 岩本裕之

    第10回 日本生物物理学関東支部会  2012.3  日本生物物理学会

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    タンパク分子の構造は,けっして均一で等方的なものではないために,冷却時には非等方的な収縮を示すと予測される。そのような物性はタンパク分子の機能や構造の安定性を理解する上で重要な知見となるが,直接調べることは難しかった。本研究では,流動配向させたウシ脳微小管を15秒で37から17℃まで急速冷却し,同時にX線繊維回折法により構造変化を追跡することに成功した。GDPチューブリンからなる微小管は,低温で脱重合する直前まで,長軸方向・直径方向に,それぞれ60,220×10-6/Kの熱膨張係数を示し,非等方的な冷却収縮を示すことがわかった。

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  • Direct imaging of the world of low Reynolds number hydrodynamics

    The Fifth International Symposium on Aero Aqua Bio-mechanisms, ISABMEC 2012 Taipei  2012 

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  • Mechanical properties of flagellar motility: Prokaryotes versus Eukaryotes

    Kamimura, S

    FASEB Summer Research Conference, Saxtons River, Vermont  2010.7 

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  • A new function of primary cilia: a cell-signaling enhancer

    Takao, D, Kamimura, S

    FASEB Summer Research Conference, Saxtons River, Vermont  2010.7 

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  • Mechanical properties of flagellar motility: Prokaryotes versus Eukaryotes

    FASEB Summer Research Conference, Saxtons River, Vermont  2010 

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  • A new function of primary cilia: a cell-signaling enhancer

    FASEB Summer Research Conference, Saxtons River, Vermont  2010 

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009.11 

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  • X-ray fiber diffraction analysis of axoneme and microtubules structure using a novel shear-flow fiber-aligning technique.

    上村慎治

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009.11 

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    4th International Symposium on Aero Aqua Bio-Mechanisms., 2009.8.30, Shanghai(China)  2009.8 

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    4th International Symposium on Aero Aqua Bio-Mechanisms., 2009.8.30, Shanghai(China)  2009 

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  • Mechanical properties of flagellar motility: Prokaryotes versus eukaryotes.

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009 

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  • X-ray fiber diffraction analysis of axoneme and microtubules structure using a novel shear-flow fiber-aligning technique.

    International Workshop Dynein 2009, 2009.11.3, Kobe(Japan)  2009 

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  • A new microscope optics for laser darkfield illumination applied to high precision measurement of specimen displacement.

    Noda, N, Kamimura, S

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008.3 

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    With conventional light microscopy, precision in the measurement of the displacement of a specimen depends on the signal-to-noise ratio when we measure the light intensity of magnified images. This implies that, for the improvement of precision, getting b

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  • Dynein arm arrangement in sea-urchin sperm flagellar axonemes revealed by small-angle X-ray diffraction analysis.

    Kamimura, S, Iwamoto, H

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008.3 

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  • A new microscope optics for laser darkfield illumination applied to high precision measurement of specimen displacement.

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008 

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  • Dynein arm arrangement in sea-urchin sperm flagellar axonemes revealed by small-angle X-ray diffraction analysis.

    Biophys. Soc. 52nd Ann. Meeting/ 16th IUPAB Int. Biophys. Congress/Biophysical Society  2008 

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  • Dynamic change of axonemeal structure and dynein arm arrangement in sea-urchin sperm flagella revealed by small-angle X-ray diffraction analysis.

    Kamimura, S, Iwamoto, H

    FASEB Summer Research Conference (The Biology of Cilia & Flagella)/FASEB organization  2007.8 

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  • Dynein arm arrangement in flagellar axonemes and its dynamic change revealed by small-angle X-ray diffraction analysis.

    Kamimura S, Iwamoto, H

    The Molecular Motor Conference/Fujihara Foundation of Science  2007.8 

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  • Dynamic change of axonemeal structure and dynein arm arrangement in sea-urchin sperm flagella revealed by small-angle X-ray diffraction analysis.

    FASEB Summer Research Conference (The Biology of Cilia & Flagella)/FASEB organization  2007 

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  • Dynein arm arrangement in flagellar axonemes and its dynamic change revealed by small-angle X-ray diffraction analysis.

    The Molecular Motor Conference/Fujihara Foundation of Science  2007 

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  • X-ray diffraction from dyenin motors and microtubules in sea -urchin sperm flagellar axonemes.

    Kamimura, S

    生物物理,日本生物物理学会  2006.11 

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  • 3-D measurement of the nanometer displacement of microspheres under optical mixcroscope

    Noda, N, Kamimura, S

    生物物理,日本生物物理学会  2006.11 

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  • 新しい流動配向法によるウニ精子鞭毛X線回折の観察

    上村 慎治, 杉山 貴紹, 杉本 泰伸, 若林克三

    社団法人日本動物学会第77回大会,日本動物学会  2006.9 

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  • X-ray diffraction from dyenin motors and microtubules in sea -urchin sperm flagellar axonemes.

    2006 

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  • 3-D measurement of the nanometer displacement of microspheres under optical mixcroscope

    2006 

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  • 鞭毛軸糸高速微小振動のナノメーター計測

    野田直紀, 上村慎治

    生物物理,日本生物物理学会  2005.11 

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  • AFMを用いたウニ精子鞭毛軸糸における高速微小振動の検出

    榊原肇

    生物物理,日本生物物理学会  2001.10 

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  • 急峻なATP濃度上昇に伴うウニ精子除膜鞭毛モデルの運動

    谷 知己, 上村慎治

    生物物理,日本生物物理学会  1996.11 

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Works

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Awards

  • ZOOLOGICAL SCIENCE Award 2011

    2011.9   Zoological Socisty of Japan   Single-Cell Electroporation of Fluorescent Probes into Sea Urchin Sperm Cells and Subsequent FRAP Analysis

    Daisuke Takao, Shinji Kamimura

  • ZOOLOGICAL SCIENCE Award 2011

    2011  

  • 井上研究奨励賞

    1985.12   井上科学振興財団  

    上村慎治

Research Projects

  • 微小管内チューブリン分子ラセン配置の柔軟性と可塑性

    2019.4 - 2021.3

    基盤研究(C)(一般) 

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    Grant amount: \4290000 ( Direct Cost: \4290000 、 Indirect Cost: \990000 )

    微小管は、真核生物のさまざまな細胞機能に深く関わる細胞骨格である。シンクロトロンの高輝度X線源を使ったX線繊維回折法を使って、微
    小管構造の柔軟性・可塑性を正確に定量的な評価を行うことで、微小管の生理機能をより深く理解できると共に、抗がん剤としての微小管安定
    化剤の薬理効果を客観的に数値化できる利点がある。これまでの電子顕微鏡技術ではわからなかった情報、動的な構造変化、その変化の時定数
    、構造の熱ゆらぎの評価を行う新手法として確立することで、抗がん剤の網羅的な探索研究を推進する。

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  • Clarification of the coupling mechanism between polymerization and GTP hydrolysis by using tubulin mutant

    Grant number:17H03668  2017.4 - 2020.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)  Institute of Physical and Chemical Research

    Muto Etsuko

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    Grant amount: \17030000 ( Direct Cost: \13100000 、 Indirect Cost: \3930000 )

    Nucleation of microtubule is essential for cellular activities, but its mechanism is unknown because of the difficulty involved in capturing rare stochastic events in the early stage of polymerization. Combining the rapid flush negative stain electron microscopy and kinetic analysis, we demonstrated that the formation of straight oligomers with critical size is essential for nucleation. Both GDP- and GTP-tubulin assemble the single-stranded oligomers with a broad range of curvature, but upon nucleation of GTP-tubulin, the distribution of curvature is shifted to produce a minor population of straight oligomer. Our results support a model in which GTP binding generates a minor population of straight oligomers compatible with lateral association and further growth to microtubules. Our study suggests that cellular factors involved in nucleation promote it via stabilization of straight oligomers.

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  • Analysis of microtubule structure dynamics by X-ray fiber diffraction.

    2016.4 - 2019.3

    文部科学省  科学研究費補助金(日本学術振興会・文部科学省) 

    上村慎治

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    Grant type:Competitive

    Grant amount: \3800000

    "X-ray fiber diffraction is one of the most direct techniques to analysed the structure of biological fibers and molecule in medium, i.e., a native state under physiological state mimicking cytosol conditions. Using our own aligning method (Sugiyama et al., 2009; Oiwa et al., 2009; Kamimura et al., 2016) we are now describing how the microtubule structure (tubulin axial repeat, mean diameter, protofilament numbers) are dynamically modified depending on chemical (cation, tubulin binding chemicals etc) and physical (temperature, shear forces etc) conditions."

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  • 細胞骨格動態の温度特性解析

    2015 - 2018

    基礎科学研究 

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    Grant type:Competitive

    哺乳類脳微小管は、非常に高い温度感受性を有する点で、他の細胞骨格とは大きく性質を異にしている。また、GTP加水分解状態によっては、負の熱膨張係数(NTE)を持つ点も、当研究室の仕事から明らかになってきた。溶液条件で、この温度特性がどのように変化するのがを解明することで、微小管の特異的な低温感受性の生物学的な意味を理解することを目指す。

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  • Temperature dependent dynamics of cytoskeletal components in eukaryote cells.

    2015 - 2018

    Basic Science Research Program 

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    Grant type:Competitive

    Microtubules are key components of the cytoskeleton in eukaryotic cells. The dynamics between assembled microtubules and free tubulin dimers in the cytoplasm is closely related to the active shape changes of microtubule networks. One of the most unique features of microtubules is its quite high sensitivity to lowered temperatures. Mammalian brain microtubules disassemble in second by lowering the medium temperature below 20ºC. Other interesting properties of microtubules are negative-thermal expansion depending on the condition of GTP-hydrolysis by tubulin dimers. To understand the detailed mechanism of such temperature-sensitivity, we are analyzing the conformation changes of tubulin-molecules within microtubule by X-ray fiber diffraction.

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  • チューブリン・微小管の動的構造解析を利用した抗ガン作用物質の網羅的検索

    2014 - 2018

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  • Efficient survey of anti-cancer drugs by a novel X-ray fiber diffraction technique to analyze dynamic configuration changes of tubulin molecules within native microtubules.

    2014 - 2018

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  • X線繊維回折法を使った微小管構造に対する諸薬剤の効果

    2012 - 2017

    ライフサイエンス基礎科学研究 

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    微小管を安定化させるPaclitaxel(taxol)は、通常、微小管内のtubulin dimerに作用し、protofilament間の結合を強めることによって、微小管の脱会合が抑えられると考えられる。これまでの研究から、tubulinの縦方向の周期が長くなり、平均protofilament数が徐々に減少するとの報告もある。本研究では、BL45XU-SAXS(SPring-8)でのX線繊維回折法によって、構造の経時変化を分単位で追跡することで、動的な構造変化がわかった。

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  • Tubulin fine structures in native microtubules revealed by X-ray fiber diffarction

    2012 - 2017

    Basic Science Research on Life Science 

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    Grant type:Competitive

    Microtubules (MT) are key components of the cytoskeleton in eukaryotic cells. One of the most undamental questions is how MT dynamics is associated with the molecular conformation of tubulin within MTs. To address the issues, we applied our new technique for the rapid shear-flow alignment of biological filaments, which enabled us to acquire X-ray fiber diffraction data in seconds from oriented native MTs under various physiological conditions. We found that the mean periodicity of tubulin dimers in MTs were elongated from 3.85 to 3.95 nm in 1 min. Diameter changes appeared to be occurring in a 10-20 min timecourse. It is suggested paclitaxel induces quick elongation of tubulin dimers in MTs.

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  • 3次元ピコメートル計測法による軸糸ダイニン動態の解析

    2007.8 - 2009.3

    文部科学省  科学研究費補助金(日本学術振興会・文部科学省)-特定領域研究 

    上村慎治

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    Grant type:Competitive

    Grant amount: \5000000

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  • 生理学的な条件下での鞭毛・微小管の構造解析

    2007 -  

    ライフサイエンス基礎科学研究 

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    Grant type:Competitive

    新規の流動配向法を開発し、生体の繊維構造(微小管・軸糸・アクチン繊維)の動的な変化がはじめて追跡できるようになった。この手法で、これまで不明であった生体繊維の機能解明を目指している。

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  • Structure analysis of axonemes and microtub ule under physiological conditions.

    2007 -  

    Basic Science Research on Life Science 

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    Grant type:Competitive

    Our novel technique to align biological fiber in a physiological buffer medium enabled us to start new investigation on the dynamic chnages of various biological fibers. We are now revealing new features of biologically important fiber structures.

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  • Precise micro-flow analyzes for the insight into the swimming mechanism of microorganisms

    Grant number:13450096  2001 - 2004

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)  The University of Tokyo

    KAMIMURA Shinji, TAKANO Yasunari, KOBAYASHI Shunichi, SUDA Hitoshi, SHINOHARA Ken-ichi

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    Grant amount: \13000000 ( Direct Cost: \13000000 )

    The project started in order to invent a new micro-machine mechanism through the studies of mimetics or understanding microorganism swimming motion under low Reynolds number conditions. Kamimura, the leader of the project, improved a new real-time analyzer of flagellar beat frequency using a novel photo-sensing device. He used the equipment to study the real-time change of flagellar beat frequency of sea-urchin or trout spermatozoa, as-well as the mean velocity of random particle Brownian motion under an optical microscope. He also invented a new method to observe sperm motion in a very thin (<100nm) water film. Takano executed the mathematical simulations of micro-flows. He compared the real bending rigidity of Vibrio flagella with that obtained by his own estimation with Kirchhoff Rod Theory as well as by the calculation of bending and twisting moment of the structures. He also showed the morphology variation of flagellar shaft of Salmonella flagella that are composed of protofilaments of two (R/L) types. Calculation of real bacterial motion is now tried by using slender-body theory for flagella and boundary element methods for bacterial cell bodies. Kobayashi has done computer and model simulation of eukaryotic flagellar motility. His computer simulation showed the propulsive velocity of flagella depends on intrinsic resistance for filament shearing and bending. The simulated model was practically tried in large scale models, where elastic fins (microtubules) were slid to each other by magnetic power units mimicking the sliding activity of dynein motor units. Suda executed studies on molecular mechanism of force generation by molecular motors. He first showed sliding motion could be mimicked by a liquid droplet placed on a surface when it had a surface tension gradient. He also executed the analysis how myosin (myosin coated surface) generates force along with its configuration changes. Using the same experimental system he analyzed load-dependency of surface attractant and repulsive forces. Shinohara started studies on new functional π-conjugated polymers. He showed how new STM/AFM techniques were valuable and powerful tools in the field of polymer sciences. He observed a single polymer molecule and showed its spectral fluctuation was coupled with Brownian motion in a molecular scale.

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  • Detection and manipulation of single bio-motor molecules : Technical innovation and applications.

    Grant number:09279104  1997 - 2000

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Priority Areas (A) 

    KAMIMURA Shinji, SHIMAMOTO Nobuo, YANAGIDA Toshio, KINOSHITA Kazuhiko, OIWA Kazuhiro, UYEDA Taro q.p.

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    Grant amount: \195400000 ( Direct Cost: \195400000 )

    During grant support for the project we executed several different kinds of research works on the biophysical and biochemical features of bio-motors. In particular our efforts were paid to develop novel techniques that enable us to approach directly to single molecular events. K.K. developed a novel technique to detect molecular orientation from fluorescence polarization, which was applied to observe helical motions of actin filaments on myosin fibers. T.Y. improved his technique to detect a single ATP molecule during force development by myosin and the applied experiments showed ca.100 ms of delay for the force generation after ATP binding suggesting an existence of energy-storing state of myosin. He also improved micro glass-needle technique to analyze force-velocity relationship of a single myosin under various experimental conditions. S.K. developed a new technique to record FRET of single molecules with real time resolution (ca.10 Hz time resolution). New trials using various kinds of other bio-motors were also carried out during the project. N.S. showed that sliding motion of E. coli trp-repressor on DNA strands enhanced its affinity towards specific DNA sequences and has started single-molecule analysis. Inter-molecular collision among RNA polymerases was also shown by N.S. to be very crucial for the initial formation of short RNA fragments. K.K found that the rotation steps of F1-ATPases being labeled with fluorescent actin was just 120 degrees and the observed speed was as high as 100 Hz. Although the following approaches were not such experiments using single bio-motors but new studies on various types of motor proteins were carried out. S.K. applied a caged-ATP technique to analyze force develapment by axonemal dynein and showed that dynein-ADP was the force-generating intermediate of during ATP hydrolysis. K.O. analyzed detailed features of inner dynein of Chlamydomonas. He found that dynein subspecies f was a motor molecule with an exceptionally high duty-ratio and that subspecies c was a processive motor. T.Q.P.U. found co-operativity between two head domains of myosin from the analysis of single-headed myosin motors. He also found cross bridge could be highly stabilized by G680V mutation in Dictyostelium myosin. Other novel techniques were developed during the project, e.g. novel fluorescent ATP analogues to detect the conformation changes of myosin (K.O.), an artificial system to reconstruct the active directional flow by bio-motors (T.Q.P.U.), a new AFM technique using sensitive-probes to detect surface charge differences (T.Y.), AFM detection of bio-oscillatory motion (S.K.) should be still preliminary but further studies for the applications should be awaited.

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  • Structure-Based Understanding of Diversity and Similarity of Cellular Motors

    Grant number:09279101  1997 - 2000

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Priority Areas (A)  The University of Tokyo

    SUTOH Kazuo, TOYOSHIMA Yoko, KAMIYA Ritsu, KAMIMURA Shinji, OSAWA Fumio, EBASHI Setsuro

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    Grant amount: \126600000 ( Direct Cost: \126600000 )

    This research was aimed at elucidating the characteristic functional properties of Motor proteins are involved in a variety of cellular processes such as cell locomotion, cell division, phagocytosis, endocytosis and vesicle transport. These motor proteins consist of three super-families ; myosin, kinesin and dynein. Although all of these proteins have the common function to change the chemical energy released by ATP hydrolysis to mechanical energy such as filament sliding and force, their in vitro and in vivo motor functions are surprisingly diverse. We have organized the reseairch project to understand the molecular mechanism of the diverse motor functions. To carry out the project, the following three research groups have been organized. (1) Searching for new motor proteins. (2) Dynamics of motor functions. (3) Single molecule imaging and manipulation. Besides these organized groups, individual researchers have been also recruited. To coordinate these researches, we have organized a meeting and a international conference each year. We have also devised various ways to enhance collaboration among the researchers in this project. This 4 year project has finally produced large number of important publications including many collaborative papers to which a number of members of this project have contributed.

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  • Comprehensive Research for Effective Promotion of Zoology in Japan

    Grant number:08304046  1996 - 1998

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)  Institute of biologival sciences, University of Tsukuba

    OKADA Masukichi, KAMIMURA Shinji, KANZAKI Ryohei, SHICHIDA Yoshinori, KAWAMURA Kazuo, ABE Shin-ichi

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    Grant amount: \5100000 ( Direct Cost: \5100000 )

    (1) In fall of 1996, information about activities on the issue of science education was obtained by means of a questionnaire survey among heads of all branches of Zoological Society of Japan. In March of 1997, founding of Biology Union was proposed to aim exchange of informations, appeals and enlightenments toward external world.
    (2) 1997, homepage committee was organized as a subcommittee of the present one. The address of the homepage opened is http : //wwwsoc.nacsis.ac.jp/zsj/index-j.html. (3) In spring of 1997, a questionnaire survey on a review system of grants for scientific research was performed. In Oct. first forum on grants for scientific research was performed in Nara : Dr. C.Kai (Tokyo Univ.) and Mr. K.Endo (formerly the Ministry of Education, Culture, and Sports of Japan) presented lectures on the grant system. In May of 1998, second forum on grants for scientific research was performed in Yonago : Dr. M.Hayashi (Ochanomizu Univ.) and Mr. K.Miyajima (the Ministry of Education, Culture, and Sports of Japan) presented lectures on comparison of grant system for scientific research between Japan and United States. In Sep. third forum on grants for scientific research was performed in Hiroshima : Dr. N.Taniguchi (Osaka Univ.) and Mr. K.Endo (Japanese Society for Promotion of Science) and Dr. K.Masumoto (the Science Council of Japan) were invited as panelists to discuss about the issues about appropriate number of reviewers and scale of projects, and release of the judgements to applicants.
    (4) In Apr. of 1998, a proposal for promotion of zoology in Japan was presented : Enlightenments toward students, pos docs and teachers, improvements of several points to activate the annual meeting of Zoological Society of Japan, founding of P.R.committee for intensive P.R.acitivity in and out of the zoological society , and support of research activities by foreign students toward promotion of zoology in Asia, were proposed. (5) In Jan. of 1999, final meeting was held in Kyoto to summarize the activities of the third committe for future planning and discuss several issues remained. Proposal to the Ministry of Education, Culture and Sports of Japan on grant system for scientific research were also discussed.

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  • New investigation of intermolecular interaction of proteins using picometer precision microscope.

    Grant number:07558096  1995 - 1997

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (A)  The University of Tokyo

    KAMIMURA Shinji

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    Grant amount: \13600000 ( Direct Cost: \13600000 )

    In the present study we developed a new system of optical microscope having horizontal axis placed on an optical bench. There are many advantages to use the horizontal setup for the microscope, i.e.mechanical stability, flexible magnification using variable tube length of the microscope. Flexible setup for any other optical elements to be placed between objective and ocular lenses since there is no tube and stable micromanipulation is also possible. After improving time-resolution of the electronics for position detector, nm-pm precision of the measurement of specimen displacement with l0kHz rate was revealed to be possible. Two kinds of applications using the apparatus were carried out during the project. One is a precise analysis of the oscillatory motion of reactivated flagellar axonemes of sed-urchin spermatozoa. There revealed to be two distinct phases of motion ; a plateau phase which period depends on ATP concentration, and a quick sliding phase. The former and the latter would be related to ATP rebinding and dynein-tubulin interacting phase (power stroke state), respectively. The second experiment was to analyze microtubule sliding and force development during rapid UV-photolysis of caged-ATP.The observation suggest vanadate-insensitive dynein-ADP complex would be corresponding to the power-stroke intermediate of dynein.

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  • Study on kinesin motility with a nm-pm precision technique.

    Grant number:06044064  1994    

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for international Scientific Research  the University of Tokyo

    KAMIMURA Shinji, TRINCZEK Bernhard, MANDELKOW Eckhard

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    Grant amount: \5200000 ( Direct Cost: \5200000 )

    Microtubule is one of the most important cytoskeletal components which support many cell functions including intra-cellular active transportation, cell motility and cell-shapes. We have carried out two main research works during this project. The first, a new assay system was developed to check the motile activity of kinesin-head domains. In this work we have compared the activity with that of native kinesin purified from pig brain. It was suggested that motor domain with 340 amino acid (from N-terminal) purified from transformed E.coli has the motion velocity of about one tenth of native kinesin. This new assay system can be applied to test any other different type of kinesin-heads. This work was done mainly in the laboratory in Max-Plank Institute in Hamburg. The second work was to improve the microscope system to measure a fine motion under optical microscope. Especially during the project we have improved the mechanical stability of optical microscope and measuring precision. Conventional microscopes have been designed to improve the quality of images but mechanical stability is not sufficient for our purpose to measure the nm-scale movement. We have designed a new optical microscope which has a horizontal optical axis. With combination of high-intensity laser-light source we have succeeded to get around 10 pm precision with better than 100kHz time resolution. The new microscope system will be used to analyze intra-molecular interaction between MAPs /motor proteins and microtubule proteins. The work of microscope improvement has been done mainly in the laboratory of Japan.

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  • 鞭毛軸系高速振動現象における力学・化学カップリングの解析

    Grant number:04740399  1992    

    日本学術振興会  科学研究費助成事業  奨励研究(A)  東京大学

    上村 慎治

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    Grant amount: \1000000 ( Direct Cost: \1000000 )

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  • 滑り運動の多様性と統一性

    Grant number:01657001  1989    

    日本学術振興会  科学研究費助成事業  重点領域研究  名古屋大学

    神谷 律, 豊島 陽子, 茶圓 茂, 真行寺 千佳子, 上村 慎治, 今栄 康雄

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    Grant amount: \22000000 ( Direct Cost: \22000000 )

    本研究は生体運動の共通原理を探るという立場から、細菌鞭毛、真核細胞鞭毛、原形質流動など筋肉以外の生体運動の機構を追及した。方法として、運動に関与した構造一つづつの動態を直視することを重視した。豊島は単離した骨格筋の太い繊維上をアクチン繊維が滑走するという試験管内運動系を開発し、アクチン繊維が双極性のミオシン繊維のどちらの半分とでも相互作用して滑ることができるという現象を観察した。これはアクチンとミオシンの相互作用にはこれまでの想像を超えた自由度があることを意味している。今栄は細菌鞭毛モーターの共有結果性阻害剤を開発し、その存在下で1個のモーターの挙動を追跡したところ、回転運動が段階的に阻害されていくことを見いだした。運動に関与したイオンチヤンネル1つづつが阻害されていく過程を捉えたものと考えられる。真行寺は真核生物の鞭毛に外力を加えてその応答を見る研究を行った。その結果、鞭毛の打つ面を人為的に回転させることができることを発見した。微小管滑り運動の調節という観点からきわめて興味深い現象である。上村は空間精度1nm、時間分解能1msecで運動を検出できる装置を開発し、上村と神谷はそれによって鞭毛軸糸内の微小な構造の揺らぎを検出する試みを行った。その結果、一見運動を停止している軸糸が、ATP存在下で、振幅1nm、周期300Hzという高速微小振動を行う現象を発見した。その振動数は個々のダイニンの動きを反映している可能性がある。今後、この現象の意味が明らかになれば、運動の分子機構の解明に大いに役立つ実験系になるであろう。この方法をアクチンーミオシン系など他の生体運動系に適用し、微少な運動の揺らぎをさまざまな条件下で検出/解析することにより、化学ー力学変換過程に関する新情報が得られる可能性がある。現在そのような試みが茶圓によって準備されている。来年度からの成果が待たれる。

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  • ベン毛などの運動の制御機構・基本構造・力学的な特性が生物進化でどんな意味付けができるのか探っています。

    1984 -  

    基礎科学研究 

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    Grant type:Competitive

    真核生物の鞭毛・繊毛の運動の制御機構・基本構造・力学的な特性を調べ、この運動機構が、生物進化の上でどのような意味付けができるのか探っています。特に、真核生物は何故すべてが鞭毛・繊毛を持つに至ったのか(その子孫であるのか)、その利点と欠点を理解することで、真核生物の誕生の過程が解明できると考えています。

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  • Motility mechanisms and mechanical properties of eukaryote falagellum and its evolutional aspects.

    1984 -  

    Basic Science Research Program 

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    Grant type:Competitive

    We are now investigating the mechanisms of regulation, structural and mechanical properties of eukaryotic cilia and flagella, expecting such studies would lead us to understand the evolutionary history of eukaryotes. A major question is why all the eukaryotes (or our ancient organisms) had to have cilia and flagella. We can know the answer to this question after the fundamental understanding of the advantage and disadvantage of the motile system of cilia and flagella.

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  • 2024   Animal Molecular Physiology   Department

  • 2024   Experimental Course of Cellular Biochemistry   Department

  • 2024   Graduation Thesis Ⅰ   Department

  • 2024   Graduation Thesis Ⅱ   Department

  • 2024   Basic Biology   Department

  • 2024   Applied Biology   Department

  • 2024   Basic Biotechnology   Department

  • 2024   English for Bioscience - Advanced Course 1   Department

  • 2024   English for bioscience - advanced course 2   Department

  • 2024   English for bioscience - basic course   Department

  • 2024   Biological Experiments 1   Department

  • 2024   Natural History Fieldwork   Department

  • 2024   Molecular Biophysics   Graduate school

  • 2024   Doctoral Research Ⅰ   Graduate school

  • 2024   Doctoral Research Ⅱ   Graduate school

  • 2024   Doctoral Research Ⅲ   Graduate school

  • 2024   Doctoral Research Ⅳ   Graduate school

  • 2024   Doctoral Research Ⅴ   Graduate school

  • 2024   Doctoral Research Ⅵ   Graduate school

  • 2024   Special lecture on Biological Sciences   Graduate school

  • 2024   Master's Research Ⅰ   Graduate school

  • 2024   Master's Research Ⅲ   Graduate school

  • 2024   Master's Research Ⅱ   Graduate school

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Committee Memberships

  • 2009 -  

    (社)日本動物学会   評議員