Post-fusion structural changes and their roles in exocytosis and endocytosis of dense-core vesicles.

Post-fusion structural changes and their roles in exocytosis and endocytosis of dense-core vesicles.
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DOI:
10.1038/ncomms4356
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发表时间:
2014
影响因子:
16.6
通讯作者:
Wu, Ling-Gang
Wu, Ling-Gang
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Chiang, Hsueh-Cheng;Shin, Wonchul;Zhao, Wei-Dong;Hamid, Edaeni;Sheng, Jiansong;Baydyuk, Maryna;Wen, Peter J.;Jin, Albert;Momboisse, Fanny;Wu, Ling-Gang

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囊泡与质膜融合产生Ω形膜轮廓。它的孔被认为是扩张直到变平(完全塌陷),然后是经典的内吞作用以取回囊泡。或者,孔可能关闭(吻和运行),但触发机制和内吞作用仍然知之甚少。在这里,使用共聚焦和STED成像致密核心囊泡,我们发现,融合产生的Ω-配置文件可能会扩大或缩小,同时保持囊泡膜蛋白。融合产生的Ω-分布的闭合(其产生各种大小的囊泡)是介导约1-30 s内的快速和缓慢内吞作用的主要机制。强钙内流触发动力蛋白介导的闭合。弱钙内流不会促进闭合,但通过收缩而不是完全塌陷促进Ω-曲线与质膜的合并。这些结果建立了称为Ω-外吞-内吞作用的模型,其中融合产生的Ω-分布可以收缩以与质膜合并,尺寸改变,或尺寸改变然后响应于钙而闭合,这是取回致密核心囊泡的主要机制。
Vesicle fusion with the plasma membrane generates an Ω-shaped membrane profile. Its pore is thought to dilate until flattening (full-collapse), followed by classical endocytosis to retrieve vesicles. Alternatively, the pore may close (kiss-and-run), but the triggering mechanisms and its endocytic roles remain poorly understood. Here, using confocal and STED imaging of dense-core vesicles, we find that fusion-generated Ω-profiles may enlarge or shrink while maintaining vesicular membrane proteins. Closure of fusion-generated Ω-profiles, which produces various sizes of vesicles, is the dominant mechanism mediating rapid and slow endocytosis within ~1–30 s. Strong calcium influx triggers dynamin-mediated closure. Weak calcium influx does not promote closure, but facilitates the merging of Ω-profiles with the plasma membrane via shrinking rather than full-collapse. These results establish a model, termed Ω-exo-endocytosis, in which the fusion-generated Ω-profile may shrink to merge with the plasma membrane, change in size, or change in size then close in response to calcium, which is the main mechanism to retrieve dense-core vesicles.
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