Characterization of docking and fusion of synaptic-like microvesicles in PC12 cells using TIRFM  

作　　者：ZHOU Wei ZHU Dan LIANG Tao LI ChenHong WU ZhengXing 

机构地区：[1]Key Laboratory of Molecular Biophysics/Ministry of Education, and Joint Laboratory of Institute of Biophysics, Huazhong University of Science and Technology, Wuhan 430074, China

出　　处：《Chinese Science Bulletin》2007年第22期3089-3096,共8页

基　　金：Supported by the National Natural Science Foundation of China (Grant Nos. 30670502 and 30470646) ;the Major State Basic Research Program of China (Grant No. 2006CB503908)

摘　　要：Neurotransmitters are released by the fusion of synaptic vesicles with presynaptic membrane,which has been extensively studied. The analysis of single vesicle fusion kinetics reveals that there exist fusion modes of "kiss and run" and "kiss and stay" which may be favored by neurons especially during strong firing beside full fusion. Pre-fusion steps of translocation,docking and priming along the exo-cytotic pathway play important roles in neurotransmitter release and its regulation. In the present report,we used dual-color imaging of VAMP2-pHluorin and VAChT-TDimer2 under total internal reflection fluorescence microscope(TIRFM) to monitor the docking and fusion of synaptic-like microvesicles(SLMVs) in PC12 cells stimulated by high K+. Our results show that "kiss and run" is a dominative fu-sion mode in PC12 cells under high K+-challenge,and the dwell time of SLMVs is prolonged by the high K+ stimulation that suggests an enhancement of vesicle priming.Neurotransmitters are released by the fusion of synapUc vesicles with presynaptic membrane, which has been extensively studied. The analysis of single vesicle fusion kinetics reveals that there exist fusion modes of ＂kiss and run＂ and ＂kiss and stay＂ which may be favored by neurons especially during strong firing beside full fusion. Pre-fusion steps of translocation, docking and priming along the exocytotic pathway play important roles in neurotransmitter release and its regulation. In the present report, we used dual-color imaging of VAMP2-pHluorin and VAChT-TDimer2 under total internal reflection fluorescence microscope （TIRFM） to monitor the docking and fusion of synaptic-like mlcrovesicles （SLMVs） in PC12 cells stimulated by high K^＋. Our results show that ＂kiss and run＂ is a dominative fusion mode in PC12 cells under high K^＋-challenge, and the dwell time of SLMVs is prolonged by the high K^＋ stimulation that suggests an enhancement of vesicle priming.

关 键 词：PC12细胞 微泡 分析方法 神经传递素 

分 类 号：Q25[生物学—细胞生物学]