Synaptotagmin I is necessary for compensatory synaptic vesicle endocytosis in vivo

Kira E. Poskanzer, Kurt W. Marek, Sean T. Sweeney and Graeme W. Davis ()
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Kira E. Poskanzer: University of California, San Francisco
Kurt W. Marek: University of California, San Francisco
Sean T. Sweeney: University of California, San Francisco
Graeme W. Davis: University of California, San Francisco

Nature, 2003, vol. 426, issue 6966, 559-563

Abstract: Abstract 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.

Date: 2003
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DOI: 10.1038/nature02184

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