Spatiotemporal visualization of intracellular calcium signal for regulation of sperm function

• Project/Area Number: 26840061
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: University of Tsukuba
• Principal Investigator: SHIBA Kogiku 筑波大学, 生命環境系, 助教 (70533561)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2014: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: 鞭毛運動 / 精子 / カルシウム

Outline of Final Research Achievements

This research was carried out to elucidate sperm function in the various stages of fertilization. Especially intracellular calcium signaling was focused. The following results were obtained. (1) To visualize sperm function spatiotemporally the system for sperm stimulation by using caged compounds were constructed. (2) Experiments using caged ATP revealed that calaxin is involved in propagation and switching of flagellar waveforms. (3) Soluble adenylyl cyclase critically plays a role in the regulation of flagellar motility and Ca2+-dependent regulation of flagellar waveform during chemotactic movement.

Research Products

• [Journal Article] Flagellar waveforms of gametes in the brown alga Ectocarpus siliculosus. (2016)
  • Author(s): Kinoshita N, Shiba K, Inaba K, Fu G, Nagasato C and Motomura T.
  • Journal Title: European Journal of Phycology Volume: 51 Issue: 2 Pages: 139-148
  • DOI: 10.1080/09670262.2015.1109144
  • NAID: 120006220013
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Branchial cilia and sperm flagella recruit distinct axonemal components (2015)
  • Author(s): Konno A, Shiba K, Cai C, Inaba K
  • Journal Title: PLoS One Volume: 10 Issue: 5 Pages: 10-10
  • DOI: 10.1371/journal.pone.0126005
  • Peer Reviewed / Open Access
• [Journal Article] Sperm calcineurin inhibition prevents mouse fertility with implications for male contraceptive (2015)
  • Author(s): Haruhiko Miyata, Yuhkoh Satouh, Daisuke Mashiko, Masanaga Muto, Kaori Nozawa, Kogiku Shiba, Yoshitaka Fujihara, Ayako Isotani, Kazuo Inaba, Masahito Ikawa
  • Journal Title: Science Volume: 350 Issue: 6259 Pages: 442-445
  • DOI: 10.1126/science.aad0836
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Lactic acid is a sperm motility inactivation factor in the sperm storage tubules. (2015)
  • Author(s): Matsuzaki M, Mizushima S, Hiyama G, Hirohashi N, Shiba K, Inaba K, Suzuki T, Dohra H, Ohnishi T, Sato Y, Kohsaka T, Ichikawa Y, Atsumi Y, Yoshimura T, Sasanami T.
  • Journal Title: Sci Rep. Volume: 5 Issue: 1 Pages: 17643-17643
  • DOI: 10.1038/srep17643
  • NAID: 120007128467
  • Peer Reviewed / Open Access
• [Journal Article] Chemotactic response with a constant delay-time mechanism in Ciona spermatozoa revealed by a high time resolution analysis of flagellar motility. (2015)
  • Author(s): Miyashiro, D. Shiba, K., Miyashita, T., Baba, S., Yoshida, M., and Kamimura, S.
  • Journal Title: Biology Open Volume: 4 Issue: 2 Pages: 109-118
  • DOI: 10.1242/bio.20137351
  • Peer Reviewed / Open Access
• [Journal Article] Distinct roles of soluble and transmembrane adenylyl cyclases in the regulation of flagellar motility in Ciona sperm. (2014)
  • Author(s): Shiba K, Inaba K.
  • Journal Title: Int J Mol Sci. Volume: 15 Issue: 8 Pages: 13192-13208
  • DOI: 10.3390/ijms150813192
  • NAID: 120007136613
  • Peer Reviewed / Open Access
• [Journal Article] Species and gamete-specific fertilization success of two sea urchins under near future levels of pCO2. (2014)
  • Author(s): Sung CG, Kim TW, Park YG, Kang SG, Inaba K, Shiba K, Choi TS, Moon SD, Litvin S, Lee KT, Lee JS.
  • Journal Title: Journal of Marine Systems. Volume: 137 Pages: 67-73
  • DOI: 10.1016/j.jmarsys.2014.04.013
  • Peer Reviewed
• [Journal Article] The birth of quail chicks after intracytoplasmic sperm injection (2014)
  • Author(s): Mizushima, S., Hiyama, G., Shiba, K., Inaba, K., Dohra, H., Ono, T., Shimada, K. and Sasanami, T.
  • Journal Title: Development Volume: 141 Issue: 19 Pages: 3799-3806
  • DOI: 10.1242/dev.111765
  • NAID: 120005479934
  • Peer Reviewed / Open Access
• [Presentation] Sperm calcineurin is necessary for midpiece flexibility and male fertility (2016)
  • Author(s): Haruhiko Miyata, Yuhkoh Satouh, Daisuke Mashiko, Masanaga Muto, Kaori Nozawa, Kogiku Shiba, Yoshitaka Fujihara, Ayako Isotani, Kazuo Inaba, Masahito Ikawa
  • Organizer: Biophysical Society 60th Annual Meeting
  • Place of Presentation: Los Angeles Convention Center, Los Angeles, CA, USA.
  • Year and Date: 2016-02-27
  • Int'l Joint Research
• [Presentation] Sperm-specific calcineurin is necessary for midpiece flexibility and male fertility (2016)
  • Author(s): Haruhiko Miyata, Yuhkoh Satouh, Daisuke Mashiko, Masanaga Muto, Kaori Nozawa, Kogiku Shiba, Yoshitaka Fujihara, Ayako Isotani, Kazuo Inaba, Masahito Ikawa
  • Organizer: 国際シンポジウム "生殖細胞のエピゲノムダイナミクスとその制御"
  • Place of Presentation: 京都大学百周年時計台記念館, 京都府、京都市
  • Year and Date: 2016-02-17
  • Int'l Joint Research
• [Presentation] 鞭毛非対称波の伝播におけるカラクシンの役割 (2016)
  • Author(s): 柴小菊, 馬場昭次, 藤原英史, 稲葉一男
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: キャンパスプラザ京都, 京都府、京都市
  • Year and Date: 2016-01-08
• [Presentation] 炭酸脱水酵素/CO2 を介したカレイ類系統特異的な精子運動阻害 (2015)
  • Author(s): 佐野 マリコ，柴 小菊，稲葉 一男
  • Organizer: 日本動物学会第86回大会
  • Place of Presentation: 新潟コンベンションセンター朱鷺メッセ, 新潟県、新潟市
  • Year and Date: 2015-09-17
• [Presentation] RNAseq データを用いたプロテオミクスによる海産無脊椎動物繊毛の多様性機構に関する研究 (2015)
  • Author(s): 稲葉 一男，柴 小菊，柴田 大輔，Wenjie Zuo，城倉 圭，牧野 由美子，山口 勝司，重信 秀治
  • Organizer: 日本動物学会第86回大会
  • Place of Presentation: 新潟コンベンションセンター朱鷺メッセ, 新潟県、新潟市
  • Year and Date: 2015-09-17
• [Presentation] 紅茶高分子ポリフェノールMAF によるムラサキウニ精子の運動活性化 (2015)
  • Author(s): 菊池 絢子，柴 小菊，中野 賢太郎，小澤 哲夫，稲葉 一男，沼田 治
  • Organizer: 日本動物学会第86回大会
  • Place of Presentation: 新潟コンベンションセンター朱鷺メッセ, 新潟県、新潟市
  • Year and Date: 2015-09-17
• [Presentation] 低酸素状態は乳酸産生および精子の運動停止に関与する (2015)
  • Author(s): 松崎 芽衣，柴 小菊，稲葉 一男，道羅 英夫，笹浪 知宏
  • Organizer: 日本動物学会第86回大会
  • Place of Presentation: 新潟コンベンションセンター朱鷺メッセ, 新潟県、新潟市
  • Year and Date: 2015-09-17
• [Presentation] カタユウレイボヤ精子鞭毛における内腕ダイニンのキャラクタリゼーション (2015)
  • Author(s): Osamu Kutomi, Katsutoshi Mizuno, Keiko Hirose, Lixy Yamada, Hitoshi Sawada, Kogiku Shiba, Kazuo Inaba
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学角間キャンパス, 石川県、金沢市
  • Year and Date: 2015-09-13
• [Presentation] Inhibition of sperm-specific calcineurin for male contraception (2015)
  • Author(s): Haruhiko Miyata, Yuhkoh Satouh, Daisuke Mashiko, Masanaga Muto, Kaori Nozawa, Kogiku Shiba, Yoshitaka Fujihara, Ayako Isotani, Kazuo Inaba, Masahito Ikawa
  • Organizer: Gordon Research Conference: Fertilization and the Activation of Development
  • Place of Presentation: Holderness, NH, USA.
  • Year and Date: 2015-07-19
  • Int'l Joint Research
• [Presentation] The role of soluble adenylyl cyclase in the regulation of flagellar motility in Ciona sperm (2015)
  • Author(s): Kogiku Shiba, Kazuo Inaba
  • Organizer: The 8th International Tunicate Meeting
  • Place of Presentation: Aomori City Cultural Hall, Aomori, Japan.
  • Year and Date: 2015-07-13
  • Int'l Joint Research
• [Presentation] Two novel subunits of axonemal dyneins from Ciona intestinalis, as potentially key regulators for ciliary motility (2015)
  • Author(s): Kazuo Inaba, Osamu Kutomi, Katsutoshi Mizuno, Kogiku Shiba, Keiko Hirose
  • Organizer: The 8th International Tunicate Meeting
  • Place of Presentation: Aomori City Cultural Hall, Aomori, Japan.
  • Year and Date: 2015-07-13
  • Int'l Joint Research
• [Presentation] 多機能運動装置ハプトネマが示す新規微小管系運動のメカニズム (2015)
  • Author(s): 阿閉耕平、柴小菊、中山剛、石田健一郎、稲葉一男
  • Organizer: 生体運動班会議２０１５
  • Place of Presentation: 東京都豊島区、学習院大学
  • Year and Date: 2015-01-07 – 2015-01-09
• [Presentation] ホヤ精子鞭毛の新規タンパク質DYBLUPとf/I1ダイニンとの相互作用 (2015)
  • Author(s): 久冨理、広瀬恵子、柴小菊、稲葉一男
  • Organizer: 生体運動班会議２０１５
  • Place of Presentation: 東京都豊島区、学習院大学
  • Year and Date: 2015-01-07 – 2015-01-09
• [Presentation] ホヤ精子における可溶性アデニル酸シクラーゼの役割 (2014)
  • Author(s): 柴小菊，稲葉一男
  • Organizer: 日本動物学会第85回大会
  • Place of Presentation: 仙台市、東北大学川内北キャンパス
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] カタユウレイボヤ精子鞭毛における内腕ダイニンf の単離・解析 (2014)
  • Author(s): 久冨理，水野克俊，広瀬恵子，柴田大輔，山田力志，澤田均，柴小菊，稲葉一男
  • Organizer: 日本動物学会第85回大会
  • Place of Presentation: 仙台市、東北大学川内北キャンパス
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] チューブリンポリグルタミン酸化酵素Ttll9 欠損マウス精子に見られる特異な構造異常と運動異常の解析 (2014)
  • Author(s): 紺野在，柴小菊，池上浩司，稲葉一男，瀬藤光利
  • Organizer: 日本動物学会第85回大会
  • Place of Presentation: 仙台市、東北大学川内北キャンパス
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] 乳酸はウズラ精子貯蔵管における精子の運動停止に関与する (2014)
  • Author(s): 松崎芽衣，水島秀成，柴小菊，稲葉一男，笹浪知宏
  • Organizer: 日本動物学会第85回大会
  • Place of Presentation: 仙台市、東北大学川内北キャンパス
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] The role of soluble adenylyl cyclase in the regulation of flagellar motility in Ciona sperm. (2014)
  • Author(s): Kogiku Shiba, Kazuo Inaba
  • Organizer: The 12th International Symposium on Spermatology
  • Place of Presentation: Newcastle, Australia
  • Year and Date: 2014-08-08 – 2014-08-10
• [Presentation] Molecular characterization of axonemal dyneins of sperm flagella in the Ascidian Ciona intestinalis. (2014)
  • Author(s): Kazuo Inaba, Osamu Kutomi, Katsutoshi Mizuno, Keiko Hirose, Kogiku Shiba, Lixy Yamada, Hitoshi Sawada
  • Organizer: The 12th International Symposium on Spermatology
  • Place of Presentation: Newcastle, Australia
  • Year and Date: 2014-08-08 – 2014-08-10
• [Presentation] Sperm chemotaxis is mediated by calcium extrusion via plasma membrane Ca2+-ATPase and Na+/Ca2+ exchanger. (2014)
  • Author(s): Kaoru Yoshida, Kogiku Shiba, Akira Nakashima, Ayako Sakamoto, Shigeru Matsunaga, Kazuo Inaba, Manabu Yoshida.
  • Organizer: The 12th International Symposium on Spermatology
  • Place of Presentation: Newcastle, Australia
  • Year and Date: 2014-08-08 – 2014-08-10
• [Presentation] Phylogenetic aspects of flagellar outer arm dynein. (2014)
  • Author(s): Hideo Mohri, Miyoko Kubo-Irie, Masaru Irie, Kogiku Shiba, Kazuo Inaba.
  • Organizer: The 12th International Symposium on Spermatology
  • Place of Presentation: Newcastle, Australia
  • Year and Date: 2014-08-08 – 2014-08-10
• [Book] Flagellar Mechanics and Sperm Guidance (Cosson J, Ed) (2015)
  • Author(s): Inaba K, Kutomi O, Shiba K, Cosson J.
  • Total Pages: 432
  • Publisher: Bentham Science Publishers
• [Book] 動植物の受精学-共通機構と多様性（澤田均編） (2014)
  • Author(s): 稲葉一男、柴小菊、吉田学
  • Total Pages: 340
  • Publisher: 化学同人
• [Remarks] 筑波大学下田臨海実験センター発表論文
  • URL: http://www.shimoda.tsukuba.ac.jp/ronbun.html
• [Remarks] マリンバイオ共同推進機構（JAMBIO）研究業績
  • URL: http://www.shimoda.tsukuba.ac.jp/~jambio/kenkyu.html
• [Remarks] 筑波大学下田臨海実験センター稲葉研究室発表論文
  • URL: http://www.shimoda.tsukuba.ac.jp/~inaba/publish.html
• [Remarks] 筑波大学下田臨海実験センター、発表論文
  • URL: http://www.shimoda.tsukuba.ac.jp/ronbun.html
• [Remarks] 筑波大学下田臨海実験センター稲葉研究室、発表論文
  • URL: http://www.shimoda.tsukuba.ac.jp/~inaba/publish.html