NAIST 奈良先端科学技術大学院大学 バイオサイエンス領域


Ultrafast recycling of synaptic vesicles

演題 Ultrafast recycling of synaptic vesicles
講演者 Dr.Shigeki Watanabe
Department of Cell Biology, Johns Hopkins School of Medicine
使用言語 English
日時 2016年3月8日(火曜日) 11:00~12:00
場所 Bio Large Seminar Room
内容 Neurons can fire at extremely high rates. To sustain neurotransmission, synaptic vesicles must be recycled locally at synapses. Two models for synaptic vesicle endocytosis have been put forward based on the morphological studies in frog neuromuscular junctions. Heuser and Reese proposed that endocytosis occurs via a slow mechanism using clathrin scaffolds. Ceccarelli and his coworkers proposed a fast mechanism, kiss-and-run. Since then, many studies have sought to identify the mechanism for synaptic vesicle endocytosis. However, instead of resolving the issue, conflicting evidence has accumulated over the years. The molecular studies have suggested clathrin and clathrin-associated proteins are essential. However, the kinetics studies have suggested that both forms co-exist. Our data identify a third pathway that is fast, but requires clathrin to regenerate vesicles. To investigate how endocytosis takes place, we developed a method, ‘flash-and-freeze’ fixation that couples optogenetic stimulation with rapid high-pressure freezing and captures endocytosis at millisecond temporal resolution. To our surprise, vesicle membrane is recovered via ultrafast endocytosis within 100 ms following a single stimulus. The large endocytic vesicles then fuse to form an endosome and are resolved by clathrin into synaptic vesicles in 5-6 s. When experiments are performed at 20ºC instead of 37ºC, ultrafast endocytosis fails, and clathrin regenerates synaptic vesicles directly from plasma membrane. These results suggest that recycling of synaptic vesicles is normally a rapid two-step process: ultrafast endocytosis that removes excess membrane from the surface and then clathrin-mediated biogenesis of synaptic vesicles from endosomes.
問合せ先 分子医学細胞生物学
末次 志郎 (