• Top page
  • Timetable
  • Per session
  • Per presentation
  • How to
  • Meeting Planner



Gene Expression and Translational Regulation

開催日 2014/9/11
時間 18:15 - 19:00
会場 Room J(313+314)
Chairperson(s) 奥野 浩行 / Hiroyuki Okuno (京都大学大学院医学研究科 メディカルイノベーションセンター / Medical Innovation Center, Kyoto University Graduate School of Medicine, Japan)
重本 隆一 / Ryuichi Shigemoto (IST Austria, Austria)

High resolution analysis of presynaptic protein localizations in the parallel fiber-Purkinje cell synapses

  • O1-J-6-2
  • 重本 隆一 / Ryuichi Shigemoto:1 別府 薫 / Kaoru Beppu:2 松井 広 / Ko Matsui:2 渡辺 雅彦 / Masahiko Watanabe:3 坂本 寛和 / Hirokazu Sakamoto :4 並木 繁行 / Shigeyuki Namiki :4 廣瀬 謙造 / Kenzo Hirose:4 原田 春美 / Harumi Harada:1 
  • 1:IST Austria, Austria / IST Austria, Austria 2:東北大学大学院医学系研究科 / Tohoku University Graduate School of Medicine, Japan 3:北海道大学医学研究科 / Hokkaido University School of Medicine, Japan 4:東京大学大学院医学系研究科 / The University of Tokyo, Graduate School of Medicine, Japan 

Activation of the presynaptic voltage-dependent calcium channels (VDCCs) triggers vesicular release of neurotransmitters. The function of VDCCs at the presynaptic active zone is controlled by direct or indirect interactions with various types of presynaptic molecules. We previously reported localization of P/Q-type VDCC subunit Cav2.1 making small clusters within the active zone of the parallel fiber-Purkinje cell (PF-PC) synapses (Indriati et al., 2013). Although a recent proteomic study reported hundreds of molecules associated to VDCCs at the presynaptic active zone, their precise distribution and relation to the VDCC clusters are unclear. In this study, we used freeze-fracture replica immunogold labeling to visualize the ultrastructural localizations of presynaptic molecules, RIM, Munc13-1, CASK, Neurexin and the SNARE proteins relative to the Cav2.1 clusters. Gold particles for RIM and Munc13-1 were concentrated at the presynaptic active zones of PF-PC synapses making small clusters and colocalized with Cav2.1, whereas CASK and Neurexin were distributed more diffusely in the active zone. SNARE proteins were distributed all over the presynaptic elements and not particularly concentrated in the active zone. The distance between immunogold particles for RIM and Cav2.1 was the shortest among those molecules, supporting a direct interaction of RIM with the P/Q-type calcium channels (Kaeser et al., 2011). To analyze potential modification of these distribution patterns accompanied with neuronal activity, we examined transgenic rats expressing channelrhodopsin-2 in cerebellar granule cells with 2-minute continuous light stimulation in vivo. We found that the Cav2.1 distribution changed shortly after the stimulation and recovered within 4 hours.

Indriati DW et al., Quantitative localization of Cav2.1 (P/Q-type) voltage-dependent calcium channels in Purkinje cells. J Neurosci. (2013) 33(8): 3668-3678.
Kaeser PS et al., RIM Proteins Tether Ca2+ Channels to Presynaptic Active Zones via a Direct PDZ-Domain Interaction. Cell (2011) 144(2): 282-295.

Copyright © Neuroscience2014. All Right Reserved.