Imaging single synaptic vesicles at CNS presynaptic terminals

• Project/Area Number: 15K18346
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Doshisha University
• Principal Investigator: Midorikawa Mitsuharu 同志社大学, 研究開発推進機構, 助教 (60632643)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 開口放出 / 全反射蛍光顕微鏡 / イメージング / シナプス小胞

Outline of Final Research Achievements

We used total internal reflection fluorescence microscopy at the live isolated calyx of Held terminal and visualized fusions and pre-fusion activities of single synaptic vesicles just beneath the plasma membrane.
We also measured vesicle membrane and protein retrieval at the same time and in the same cell at the calyx of Held from rats and mice, and found that there may be two distinct modes of membrane and vesicle protein retrieval depending on the activity strength. We also found an important signaling pathway to couple vesicle membrane and vesicle protein uptakes.

Research Products

• [Int'l Joint Research] マックスプランク実験医学研究所/マックスプランク生物物理化学研究所(ドイツ)
• [Journal Article] Distinct modes of endocytotic presynaptic membrane and protein uptake at the calyx of Held terminal of rats and mice. (2016)
  • Author(s): Okamoto Y, Lipstein N, Hua Y, Lin KH, Brose N, Sakaba T, Midorikawa M.
  • Journal Title: eLife Volume: 5
  • DOI: 10.7554/elife.14643
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Imaging Exocytosis of Single Synaptic Vesicles at a Fast CNS Presynaptic Terminal. (2015)
  • Author(s): Midorikawa M, Sakaba T.
  • Journal Title: Neuron Volume: 88 Issue: 3 Pages: 492-8
  • DOI: 10.1016/j.neuron.2015.09.047
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Live cell imaging of single synaptic vesicles at mammalian CNS presynaptic terminals. (2017)
  • Author(s): Midorikawa M
  • Organizer: 第94回日本生理学会大会シンポジウム
  • Place of Presentation: アクトシティ浜松(静岡県・浜松市)
  • Year and Date: 2017-03-28
  • Int'l Joint Research / Invited
• [Presentation] Dynamics of single synaptic vesicles at the presynaptic terminals of mammalian central synapses (2016)
  • Author(s): Mitsuharu Midorikawa and Takeshi Sakaba
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-07-20
  • Int'l Joint Research
• [Presentation] Calmodulin-Munc 13-1 signaling regulates the coordinated uptake of synaptic vesicle membrane and protein at the calyx of Held (2016)
  • Author(s): Yuji Okamoto, Takeshi Sakaba, and Mitsuharu Midorikawa
  • Organizer: 第93回日本生理学会大会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2016-03-22
• [Presentation] Visualizing kinetics of single synaptic vesicles at the calyx-type presynaptic terminal. (2015)
  • Author(s): Mitsuharu Midorikawa and Takeshi Sakaba
  • Organizer: 第38回 日本神経科学大会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-07-28