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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Role Of Rab27 In Synaptic Transmission At The Squid Giant Synapse
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Role Of Rab27 In Synaptic Transmission At The Squid Giant Synapse

机译：Rab27在鱿鱼巨突触中突触传递中的作用。

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摘要

Small GTPase Rab is a member of a large family of Ras-related proteins, highly conserved in eukaryotic cells, and thought to regulate specific type(s) and/or specific step(s) in intracellular membrane trafficking. Given our interest in synaptic transmission, we addressed the possibility that Rab27 (a close isoform of Rab3) could be involved in cytosolic synaptic vesicle mobilization. Indeed, preterminal injection of a specific antibody against squid Rab27 (anti-sqRab27 antibody) combined with confocal microscopy demonstrated that Rab27 is present on squid synaptic vesicles. Electrophys-iological study of injected synapses showed that the anti-sqRab27 antibody inhibited synaptic release in a stimulation-dependent manner without affecting presynaptic action potentials or inward Ca~(2+) current. This result was confirmed in in vitro synaptosomes by using total internal reflection fluorescence microscopy. Thus, syn-aptosomal Ca~(2+)-stimulated release of FM1-43 dye was greatly impaired by intraterminal anti-sqRab27 antibody. Ultrastructural analysis of the injected giant preterminal further showed a reduced number of docked synaptic vesicles and an increase in nondocked vesicular profiles distant from the active zone. These results, taken together, indicate that Rab27 is primarily involved in the maturation of recycled vesicles and/or their transport to the presynaptic active zone in the squid giant synapse.
机译：小GTPase Rab是Ras相关蛋白大家族的成员，在真核细胞中高度保守，被认为可以调节细胞内膜运输中的特定类型和/或特定步骤。鉴于我们对突触传递的兴趣，我们解决了Rab27（Rab3的紧密亚型）可能参与胞浆突触小泡动员的可能性。确实，针对鱿鱼Rab27的特异性抗体（抗-sqRab27抗体）的预末端注射与共聚焦显微镜相结合证明，Rab27存在于鱿鱼突触小泡上。注射突触的电生理研究表明，抗sqRab27抗体以刺激依赖性方式抑制突触释放，而不影响突触前动作电位或内向Ca〜（2+）电流。通过使用全内反射荧光显微镜在体外突触体中证实了该结果。因此，末端内抗sqRab27抗体极大地损害了由同胞体Ca〜（2+）刺激的FM1-43染料的释放。注射的巨型前末端的超微结构分析进一步显示，停靠的突触小泡数量减少，并且远离活动区的无坞囊泡轮廓增加。这些结果加在一起，表明Rab27主要参与回收的囊泡的成熟和/或它们向鱿鱼巨突触中突触前活性区的运输。

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