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Synaptotagmin I is necessary for compensatory synaptic vesicle endocytosis in vivo

Poskanzer, K.E., Marek, K.W., Sweeney, S.T. and Davis, G.W. (2003) Synaptotagmin I is necessary for compensatory synaptic vesicle endocytosis in vivo. Nature, 426 (6966). pp. 559-563. ISSN 0028-0836

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Neurotransmission requires a balance of synaptic vesicle exocytosis and endocytosis1. Synaptotagmin I (Syt I) is widely regarded as the primary calcium sensor for synaptic vesicle exocytosis2, 3, 4, 5, 6. Previous biochemical data suggest that Syt I may also function during synaptic vesicle endocytosis7, 8, 9, 10, 11, 12, 13, 14, 15, 16; however, ultrastructural analyses at synapses with impaired Syt I function have provided an indirect and conflicting view of the role of Syt I during synaptic vesicle endocytosis3, 8, 9, 10, 14. Until now it has not been possible experimentally to separate the exocytic and endocytic functions of Syt I in vivo. Here, we test directly the role of Syt I during endocytosis in vivo. We use quantitative live imaging of a pH-sensitive green fluorescent protein fused to a synaptic vesicle protein (synapto-pHluorin) to measure the kinetics of endocytosis in sytI-null Drosophila. We then combine live imaging of the synapto-pHluorins with photoinactivation of Syt I, through fluorescein-assisted light inactivation, after normal Syt I-mediated vesicle exocytosis. By inactivating Syt I only during endocytosis, we demonstrate that Syt I is necessary for the endocytosis of synaptic vesicles that have undergone exocytosis using a functional Syt I protein.

Item Type: Article
Institution: The University of York
Academic Units: The University of York > Biology (York)
Depositing User: York RAE Import
Date Deposited: 11 Mar 2009 13:45
Last Modified: 11 Mar 2009 13:45
Published Version: http://dx.doi.org/10.1038/nature02184
Status: Published
Publisher: Nature Publishing Group
Identification Number: 10.1038/nature02184
URI: http://eprints.whiterose.ac.uk/id/eprint/7073

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