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The Journal of Neuroscience, November 1, 2000, 20(21):7986-7993
Clathrin-Mediated Endocytosis near Active Zones in Snake Motor Boutons
Haibing Teng and Robert S. Wilkinson Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110
We have used the activity-dependent probe FM1-43 with electron microscopy (EM) to examine endocytosis at the vertebrate nerve-muscle synapse. Preparations were fixed after very brief neural stimulation at reduced temperature, and internalized FM1-43 was photoconverted into an electron-dense reaction product. To locate the reaction product, we reconstructed computer renderings of individual terminal boutons from serial EM sections. Most of the reaction product was seen in 40-60 nm vesicles. All of the labeled vesicles were clathrin-coated, and 92% of them were located within 300 nm of the plasma membrane, suggesting that they had undergone little processing after retrieval from their endocytic sites. The vesicles (and by inference the sites) were not dispersed randomly near the plane of the membrane but instead were clustered significantly near active zones. Additional reaction product was found within putative macropinosomes; these appeared to form from deep membrane invaginations near active zones. Thus two mechanisms of endocytosis were evident after brief stimulation. Endocytosis near active zones is consistent with the existence of local exo/endocytic cycling pools. This mechanism also might serve to maintain alignment of active zones with postsynaptic folds during periods of activity when vesicular and plasma membranes are interchanged.
Key words: clathrin; endocytosis; nerve terminal; neuromuscular junction; neurosecretion; optical probes; vesicle processing
Copyright © 2000 Society for Neuroscience 0270-6474/00/20217986-08$05.00/0
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