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Volume 17, Number 9, Issue of May 1, 1997 pp. 3074-3084
Copyright ©1997 Society for Neuroscience

Intracellular Acidification Reversibly Reduces Endocytosis at the Neuromuscular Junction

Received Nov. 21, 1996; revised Feb. 14, 1997; accepted Feb. 18, 1997.

Clark A. Lindgren², Dennis G. Emery¹, and Philip G. Haydon¹

¹ Laboratory of Cellular Signaling, Department of Zoology and Genetics, Iowa State University, Ames, Iowa 50011, and ² Department of Biology, Grinnell College, Grinnell, Iowa 50112

The close spatial and temporal coupling of endocytosis and exocytosis in nerve terminals has made it difficult to elucidate the mechanisms and the regulation of endocytosis per se. Despite significant advances in our knowledge of the molecules involved in endocytosis, it has not yet been possible to selectively manipulate endocytosis in nerve terminals. We report that the substitution of propionate for chloride in the saline bathing a lizard neuromuscular junction reduces internal pH and reversibly blocks activity-dependent endocytosis. When intraterminal pH is reduced by ~0.7 pH units, the uptake of FM1-43 in nerve terminals, but not activity-dependent destaining, is reduced. Normalization of intracellular pH by removing the propionate, raising extracellular pH, or adding ammonium chloride immediately restores FM1-43 uptake. Electron microscopy indicates that intracellular acidification reversibly reduces activity-dependent endocytosis in nerve terminals, because depolarization in propionate saline leads to a depletion of vesicles and the appearance of large intramembraneous infoldings.

Key words: exocytosis; endocytosis; FM1-43; synaptic vesicle; neuromuscular junction

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