@article{b909531a4a73449b96ff3e2777295acf,
title = "Synaptic vesicles transiently dock to refill release sites",
abstract = "Synaptic vesicles fuse with the plasma membrane to release neurotransmitter following an action potential, after which new vesicles must {\textquoteleft}dock{\textquoteright} to refill vacated release sites. To capture synaptic vesicle exocytosis at cultured mouse hippocampal synapses, we induced single action potentials by electrical field stimulation, then subjected neurons to high-pressure freezing to examine their morphology by electron microscopy. During synchronous release, multiple vesicles can fuse at a single active zone. Fusions during synchronous release are distributed throughout the active zone, whereas fusions during asynchronous release are biased toward the center of the active zone. After stimulation, the total number of docked vesicles across all synapses decreases by ~40%. Within 14 ms, new vesicles are recruited and fully replenish the docked pool, but this docking is transient and they either undock or fuse within 100 ms. These results demonstrate that the recruitment of synaptic vesicles to release sites is rapid and reversible.",
author = "Kusick, {Grant F.} and Morven Chin and Sumana Raychaudhuri and Kristina Lippmann and Adula, {Kadidia P.} and Hujber, {Edward J.} and Thien Vu and Davis, {M. Wayne} and Jorgensen, {Erik M.} and Shigeki Watanabe",
note = "Funding Information: We are indebted to S. Li, Q. Gan, K. Itoh, D. Lubsanjav, C. Zhang and S. Markert for help with cell culture and freezing and for stimulating discussions. We also thank M. Delanoy and B. Smith for technical assistance with electron microscopy and K. T. DiNapoli for developing R code to randomize images. We thank P. Wurzinger and C. Tomova at Leica for the design and manufacture of the middle plate, M. A. Herman for initial tests of using a capacitor for field stimulation, H. Goldschmidt for help validating the stimulation device using pHluorin imaging, and N. Livingston for help with voltage imaging. We also thank the Marine Biological Laboratory and their neurobiology course for supporting the initial set of experiments (course supported by National Institutes of Health grant R25NS063307). S.W. and this work were supported by start-up funds from the Johns Hopkins University School of Medicine, Johns Hopkins Discovery funds and the National Science Foundation (1727260), and the National Institutes of Health (1DP2 NS111133-01 and 1R01 NS105810-01A1) awarded to S.W. S.W. is an Alfred P. Sloan fellow, a McKnight Foundation Scholar and a Klingenstein and Simons Foundation scholar. E.M.J. is an Investigator of the Howard Hughes Medical Institute. G.F.K. was supported by a grant from the National Institutes of Health to the Biochemistry, Cellular and Molecular Biology program of the Johns Hopkins University School of Medicine (T32 GM007445) and is a National Science Foundation Graduate Research Fellow (2016217537). The EM ICE high-pressure freezer was purchased partly with funds from an equipment grant from the National Institutes of Health (S10RR026445) awarded to S. C. Kuo. Publisher Copyright: {\textcopyright} 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2020",
month = nov,
day = "1",
doi = "10.1038/s41593-020-00716-1",
language = "English (US)",
volume = "23",
pages = "1329--1338",
journal = "Nature neuroscience",
issn = "1097-6256",
publisher = "Nature Publishing Group",
number = "11",
}