Abstract
Key components of vesicular neurotransmitter release, such as Ca
2+ influx and membrane recycling, are affected by cytosolic pH. We measured the pH-sensitive fluorescence of Yellow Fluorescent Protein transgenically expressed in mouse motor nerve terminals, and report that Ca
2+ influx elicited by action potential trains (12.5–100 Hz) evokes a biphasic pH change: a brief acidification (∼13 nM average peak increase in [H
+]), followed by a prolonged alkalinization (∼30 nM peak decrease in [H
+]) that outlasts the stimulation train. The alkalinization is selectively eliminated by blocking vesicular exocytosis with botulinum neurotoxins, and is prolonged by the endocytosis-inhibitor dynasore. Blocking H
+ pumping by vesicular H
+-ATPase (with folimycin or bafilomycin) suppresses stimulation-induced alkalinization and reduces endocytotic uptake of FM1-43. These results suggest that H
+-ATPase, known to transfer cytosolic H
+ into prefused vesicles, continues to extrude cytosolic H
+ after being exocytotically incorporated into the plasma membrane. The resulting cytosolic alkalinization may facilitate vesicular endocytosis.
► Action potentials evoke an acidification-alkalinization sequence in motor terminals ► Alkalinization requires fusion of synaptic vesicles, and recovers during endocytosis ► Terminals alkalinize by incorporating vesicular H
+-ATPase into plasma membrane ► H
+ pumping by vesicular H
+-ATPase facilitates endocytosis of vesicle membranes
