Application of photoactivated adenylyl cyclase to study the mechanisims of neurite formation by controlling temporal and spacial intracellular cAMP level in vitro

• Project/Area Number: 21590211
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: INOUE Akihiro 東京医科歯科大学, 大学院・医歯学総合研究科, 准教授 (80322080)
• Co-Investigator(Kenkyū-buntansha): NAKATA Takao 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (50218004)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 神経突起形成 / cAMP / 細胞内シグナル伝達 / 光活性化 / 光活性化アデニル酸シクラーゼ / エレクトロポレーション / メラノサイト / 神経突起伸展 / アデニル酸シクラーゼ / PAC

Research Abstract

Potencies of the photo-activated adenylyl cyclases(PAC) from euglena and bacteria for controlling intracellular cAMP concentration in vitro were analyzed biochemically with an cAMP-ELISA as well as cell-biologically with an Epac FRET sensor probe. It was also observed that melanosomes labeled with its associated protein Rab27a fused to fluorescent proteins were immediately moved toward peripheral regions of the B16 melanoma cells in response to an increase of the intracellular cAMP. We have also tried to utilize the PACs to study the morphological development and functions of neural cells in PC12 cells and hippocampal primary culture neurons.

Research Products

• [Journal Article] Preferential binding of a kinesin-1 motor to GTP-tubulin-rich microtubules underlies polarized vesicle transport (2011)
  • Author(s): Takao Nakata, Shinsuke Niwa, Yasushi Okada, Franck Perez, and Nobutaka Hirokawa.
  • Journal Title: The Journal of Cell Biology Volume: Vol.194 Issue: 2 Pages: 245-255
  • DOI: 10.1083/jcb.201104034
  • Peer Reviewed
• [Journal Article] シグナル分子の光制御技術と神経の形態形成 (2011)
  • Author(s): 中田隆夫
  • Journal Title: ブレインサイエンス・レビュー2011 Pages: 23-36
• [Journal Article] Kinase-dead knock-in mouse reveals an essential role of kinase activity of Ca2+/calmodulin-dependent protein kinase IIalpha in dendritic spine enlargement, long-term potentiation, and learning (2009)
  • Author(s): Yamagata Y, Kobayashi S, Umeda T, Inoue A, Sakagami H, Fukaya M, Watanabe M, Hatanaka N, Totsuka M, Yagi T, Obata K, Imoto K, Yanagawa Y, Manabe T, Okabe S
  • Journal Title: Journal of Neuroscience Volume: Vol.29 Pages: 7607-7618
  • Peer Reviewed
• [Journal Article] Critical involvement of Rho GTPase activity in the efficient transplantation of neural stem cells into the injured spinal cord (2009)
  • Author(s): Numano F, Inoue A, Enomoto M, Shinomiya K, Okawa A and Okabe S
  • Journal Title: Mol Brain Volume: Vol.2 Pages: 37-37
  • Peer Reviewed
• [Journal Article] 東京医科歯科大学の教授就任にあたって (2009)
  • Author(s): 中田隆夫
  • Journal Title: 解剖学雑誌 Volume: Vol.84 Pages: 26-27
  • NAID: 10026094976
• [Journal Article] Kinase-dead knock-in mouse reveals an essential role of kinase activity of Ca2+/calmodulin-dependent protein kinase IIalpha in dendritic spine enlargement, long-term potentiation, and learning (2009)
  • Author(s): Yamagata Y, et al.
  • Journal Title: Journal of Neuroscience 29 Pages: 7607-18
  • Peer Reviewed
• [Journal Article] Critical involvement of Rho GTPase activity in the efficient transplan tation of neural stem cells into the injured spinal cord (2009)
  • Author(s): Numano F, et al.
  • Journal Title: Molecular Brain 2 Pages: 37-37
  • Peer Reviewed
• [Presentation] Preferential binding of a kinesin-1 motor to GTP-tubulin-rich microtubules underlies polarized vesicle transport (2011)
  • Author(s): T. Nakata, S. Niwa, Y, Okada, F, Perez, N. Hirokawa
  • Organizer: The American Society for Cell Biology 2011 Annual Meeting
  • Place of Presentation: Denver, Colorado, USA
  • Year and Date: 2011-12-06
• [Presentation] The kinesin-1 motor protein KIF5 recognizes microtubule lattice structure emanated from GTP-tubulin in axons as directional cue.(キネシンは、GTPチュブリンに由来する微小管格子構造を認識し、軸索の方向を認識する) Neuroscience 2011 (2011)
  • Author(s): Takao Nakata, Shinsuke Niwa, Yasushi Okada, Franck Perez, Nobutaka Hirokawa
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2011-09-15
• [Presentation] The kinesin-1 motor protein KIF5 recognizes microtubule lattice structure emanated from GTP-tubulin in axons as directional cue.キネシンは、GTPチュブリンに由来する微小管格子構造を認識し、軸索の方向を認識する (2011)
  • Author(s): Takao Nakata, Shinsuke Niwa, Yasushi Okada, Franck Perez, Nobutaka Hirokawa
  • Organizer: Neuroscience 2011(第34回日本神経科学大会)
  • Place of Presentation: 横浜
  • Year and Date: 2011-09-15
• [Presentation] Rewiring vesicle transports to probe the mechanism of polarized transports in neurons.(神経細胞内の機能分子Trafficと高次脳機能そして疾患)、Neuro2010 (2010)
  • Author(s): Takao Nakata, Nobutaka Hirokawa
  • Organizer: 第33回日本神経科学大会、第53回日本神経化学大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2010-09-03
• [Presentation] Rewiring vesicle transports to probe the mechanism of polarized transports in neurons.神経細胞内の機能分子Trafficと高次脳機能そして疾患 (2010)
  • Author(s): Takao Nakata, Nobutaka Hirokawa
  • Organizer: Neuro2010(第33回日本神経科学大会、第53回日本神経化学大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2010-09-03
• [Presentation] Cdc42の光制御法の開発と培養細胞への応用 (2010)
  • Author(s): 角元利行、中田隆夫
  • Organizer: 第115回日本解剖学会総
  • Place of Presentation: 盛岡
• [Presentation] THE ROLE OF KINASE ACTIVITY OF Ca2+/CALMODULIN-DEPENDENT PROTEIN KINASE II IN HIPPOCAMPAL SYNAPTIC PLASTICITY AND LEARNING (2009)
  • Author(s): Yoko Yamagata, Shizuka Kobayashi, Tatsuya Umeda, Akihiro Inoue, Hiroyuki Sakagami, Masahiro Fukaya, Masahiko Watanabe, Nobuhiko Hatanaka, Masako Totsuka, Takeshi Yagi, Kunihiko Oabata, Keiji Imoto, Yuchio Yanagawa, Toshiya Manabe, Shigeo Okabe
  • Organizer: The 36th International Congress of Physiological Sciences
  • Place of Presentation: Kyoto, Japan
• [Presentation] カルモジュリン依存性プロテインキナーゼII & alpha ;活性は海馬シナプス可塑性の構造学的・機能学的・行動学的発現に不可欠 (2009)
  • Author(s): 山肩葉子、小林静香、梅田達也、井上明宏、阪上洋行、深谷昌弘、渡邉雅彦、畑中伸彦、戸塚昌子、八木健、小幡邦彦、井本敬二、柳川右千夫、真鍋俊也、岡部繁男
  • Organizer: 第32回日本神経科学大会
  • Place of Presentation: 名古屋
• [Presentation] カルモジュリン依存性プロテインキナーゼII&alpha ; 活性は海馬シナプス可塑性の構造学的・機能学的・行動学的発現に不可欠 (2009)
  • Author(s): 山肩葉子
  • Organizer: 第32回日本神経科学大会
  • Place of Presentation: 名古屋
• [Patent(Industrial Property Rights)] ポリペプチド、単離された核酸、組み換えベクター、遺伝子導入キット、形質転換体、および細胞内カルシウムシグナルの調節方法 (2012)
  • Inventor(s): 石井智浩、中田隆夫
  • Industrial Property Rights Holder: 東京医科歯科大学
  • Filing Date: 2012-02-16
  • Overseas
• [Patent(Industrial Property Rights)] ポリペプチド、単離された核酸、組み換えベクター、遺伝子導入キット、形質転換体、および細胞内カルシウムシグナルの調節方法 (2012)
  • Inventor(s): 石井智浩、中田隆夫
  • Industrial Property Rights Holder: 東京医科歯科大学
  • Filing Date: 2012-02-16
  • Overseas
• [Patent(Industrial Property Rights)] ポリペプチド、単離された核酸、組み換えベクター、遺伝子導入キット、形質転換体、および細胞内カルシウムシグナルの調節方法 (2012)
  • Inventor(s): 石井智浩、中田隆夫
  • Industrial Property Rights Holder: 東京医科歯科大学
  • Filing Date: 2012-02-16