Live Imaging of membrane repair by high sensitive multi-photon laser microscope in vivo system.

2014 Fiscal Year Final Research Report

• Project/Area Number: 22390374
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: Kagawa University
• Principal Investigator: MIYAKE Katsuya 香川大学, 医学部, 准教授 (30219745)
• Co-Investigator(Kenkyū-buntansha): EGAMI Youhei 香川大学, 医学部, 助教 (80432780) ; MATSUDA Chie 独立行政法人産業技術総合研究所, バイオメディカル部門, 主任研究員 (50344099) ; KANAGAWA Motoi 神戸大学, 医学研究科, 助教 (00448044) ; TANAKA Toru 城西大学, 薬学部, 准教授 (60217049) ; FUKAI Naomi 奥羽大学, 歯学部, 教授 (60134681) ; YOSHIYAMA Masahiro 岡山大学, 歯学部, 教授 (10201071)
• Co-Investigator(Renkei-kenkyūsha): ARAKI Shinichi 香川大学, 医学部, 教授 (10202748)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Keywords: 細胞膜修復 / 細胞膜損傷 / Annexin / MICAL / dysferlin / 筋ジストロフィー / 二光子 / ライブイメージング

Outline of Final Research Achievements

Plasma membrane disruption is a common form of cell injury in mammalian tissues under physiological conditions. We expressed MICAL1-, Annexins-,MG53-, dyferlin-GFPs in culture cells or in vivo, and then subjected them to a plasma membrane disruption created by a two-photon laser. Subsequent confocal imaging with a high sensitive detector unit revealed more striking wave and faster (second time-scale) accumulation of MICAL1-GFP at the disruption site comparing to MG53 or dysferlin-GFPs, followed by actin-RFP depolymerization (second time-scale). We also observed, for the first time in culture cells or living skeletal muscle cells responding to a membrane disruption, subsequent confocal imaging revealed striking accumulation of annexins (A1, A2, A4, A5, A6, A7, S100A10, S100A11)-GFPs at the disruption site. The membrane repair mechanism is now well learned at the cellular and molecular level by multi-photon laser microscope with a high sensitive detector unit.

Free Research Field

細胞生物学・組織学・解剖学