Application of fluorescent probes to single vesicle analysis

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K14321
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Doshisha University
• Principal Investigator: Sakaba Takeshi 同志社大学, 脳科学研究科, 教授 (80609511)
• Co-Investigator(Renkei-kenkyūsha): KAWAGUCHI Shinya 京都大学, 産官学連携本部, 特定准教授 (00378530) ; MIDORIKAWA Mitsuharu 東京女子医科大学, 医学部, 准講師 (60632643)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: シナプス / 神経

Outline of Final Research Achievements

In mammalian CNS synapses, transmitter is released by fusion of synaptic vesicles to the plasma membrane. Because diameter of synaptic vesicles is small, it is difficult to look at dynamics of single synaptic vesicles. In this program, we have applied total internal reflection fluorescent microscopy to acutely-dissociated presynaptic terminals. By sparsely staining synaptic vesicles with FM1-43, we could monitor dynamics of synaptic vesicles such as fusion and tethering to the release sites at the calyx of Held synapse in the auditory brainstem as well as hippocampal mossy fiber synapse. Moreover, we found that tethering of synaptic vesicles to the release sites was not sufficient for vesicle fusion. In addition to tethering, molecular priming of synaptic vesicles was required for fusion events.

Free Research Field

神経科学