Study on mechanism of rotational movement of node cilia in the mouse embryo by ultra high voltage electron microscopy

• Project/Area Number: 24770209
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology
• Research Institution: Osaka University
• Principal Investigator: SHINOHARA Kyosuke 大阪大学, 生命機能研究科, 助教 (20527387)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 繊毛 / 発生・分化

Outline of Final Research Achievements

Determination of left-right asymmetry in mouse embryos is established by a leftward flow that is generated by clockwise rotation of node cilia. Here we show exposure to the microtubule-stabilizing drug paclitaxel/Taxol markedly changes the motion pattern and ultrastructure of node cilia. In general, the node cilia show clockwise rotation. However, Taxol-treated node cilia show rotation in the random direction. We next analyze ultrastructure of node cilia by electron tomography. the node cilia harbor a regular arrangement of nine doublet microtubules, but taxol treatment randomizes the arrangement: a couple of microtubules move from the peripheral to the center of axoneme. To clarify relation between motion and structure of cilia, we carry out computer simulation of ciliary motion based on the electron tomography data. The computer simulation reveals that the regular arrangement of doublet microtubules is essential for stable unidirectional rotation of the node cilia.

Research Products

• [Journal Article] Pih1d3 is required for cytoplasmic preassembly of axonemal dynein in mouse sperm. (2014)
  • Author(s): Dong, F., Shinohara, K., Nabeshima, R., Botilde, Y., Asai, Y., Fukumoto, A., Hasegawa, T., Matsuo, M., Takeda, H., Shiratori, H., Nakamura T., and Hamada, H.
  • Journal Title: J. Cell Biol. Volume: 204 Issue: 2 Pages: 203-213
  • DOI: 10.1083/jcb.201304076
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Asymmetric rotational stroke in mouse node cilia during left-right determination (2013)
  • Author(s): A. Takamastu, T. Ishikawa, K. Shinohara, H. Hamada
  • Journal Title: Physical Review E Volume: 87 Issue: 5 Pages: 50701-50701
  • DOI: 10.1103/physreve.87.050701
  • Peer Reviewed
• [Journal Article] Hydrodynamic phase locking in mouse node cilia (2013)
  • Author(s): A. Takamastu, K. Shinohara, T. Ishikawa, H. Hamada
  • Journal Title: Physical Review Letters Volume: 110 Issue: 24 Pages: 248107-248107
  • DOI: 10.1103/physrevlett.110.248107
  • Peer Reviewed
• [Journal Article] Cluap1 localizes preferentially to the base and tip of cilia and is required for ciliogenesis in the mouse embryo (2013)
  • Author(s): Botilde, Y., Yoshiba, S., Shinohara, K., Hasegawa, T., Nishimura, H., Shiratori, H. and Hamada, H.
  • Journal Title: Developmental Biology Volume: 381 Issue: 1 Pages: 203-212
  • DOI: 10.1016/j.ydbio.2013.05.024
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The dynamic right-to-left Translocation of Cerl2 is involved in the regulation and termination of nodal activity in the mouse node (2013)
  • Author(s): Jose Manuel Ina cio, Sara Marques, Tetsuya Nakamura, Kyosuke Shinohara, Chikara Meno, Hiroshi Hamada, Jose Anto nio Belo
  • Journal Title: PLoS One Volume: 8
  • Peer Reviewed
• [Journal Article] Cilia at the Node of Mouse Embryos Sense Fluid Flow for Left-Right Determination via Pkd2 (2012)
  • Author(s): Yoshiba S, Shiratori H, Kuo IY, Kawasumi A, Shinohara K, Nonaka S, Asai Y, Sasaki G, Belo JA, Sasaki H, Nakai J, Dworniczak B, Ehrlich BE, Pennekamp P, Hamada H
  • Journal Title: Science Volume: 338(6104):226-3 Issue: 6104 Pages: 226-231
  • DOI: 10.1126/science.1222538
  • Peer Reviewed
• [Journal Article] Fluid flow and interlinked feedback loops establish left-right asymmetric decay of Cerl2 mRNA in the mouse embryo (2012)
  • Author(s): Nakamura, T., Saito, D., Kawasumi, A., Shinohara, K., Asai, Y., Takaoka, K., Dong, F., Takamatsu, A., Belo, J.A., Mochizuki, A., and Hamada, H.
  • Journal Title: Nat.Communic Volume: 3 Issue: 1 Pages: 1322-1322
  • DOI: 10.1038/ncomms2319
  • Peer Reviewed
• [Presentation] Unexpectedly fragile architecture of nodal cilia that determine left-right asymmetry in the mouse embryo (2014)
  • Author(s): Kyosuke Shinohara
  • Organizer: Cilia 2014
  • Place of Presentation: Paris, France
  • Year and Date: 2014-11-18 – 2014-11-21