Imaging direct, dynamin-dependent recapture of fusing secretory granules on plasma membrane lawns from PC12 cells

被引:139
作者
Holroyd, P
Lang, T
Wenzel, D
De Camilli, P
Jahn, R [1 ]
机构
[1] Max Planck Inst Biophys Chem, Dept Neurobiol, D-37077 Gottingen, Germany
[2] Yale Univ, Sch Med, Howard Hughes Med Inst, New Haven, CT 06510 USA
[3] Yale Univ, Sch Med, Dept Cell Biol, New Haven, CT 06510 USA
关键词
D O I
10.1073/pnas.222677399
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
During exocytosis, secretory granules fuse with the plasma membrane and discharge their content into the extracellular space. The excicytosed membrane is then reinternalized in a coordinated fashion. A role of clathrin-coated vesicles in this process is well established, whereas the involvement of a direct retrieval mechanism (often called kiss and run) is still debated. Here we report that a significant population of docked secretory granules in the neuroendocrine cell line PC12 fuses with the plasma membrane, takes up fluid-phase markers, and is retrieved at the same position. Fusion allows for complete discharge of small molecules, whereas GFP-labeled neuropeptide Y (molecular mass approximate to35 kDa) is only partially released. Retrieved granules were preferentially associated with dynamin. Furthermore, recapture is inhibited by guanosine 5'-[gamma-thio]triphosphate and peptides known to block dynamin function. We conclude that secretory granules can be recaptured immediately after formation of an exocytotic opening by an endocytic reaction that is spatially and temporally coupled to soluble N-ethylmaleimide-sensitive factor attachment protein receptor(SNARE)-dependent fusion, but is not a reversal of the fusion reaction.
引用
收藏
页码:16806 / 16811
页数:6
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