Physiological and regenerative acetylcholine release from motor nerve: differential inhibitions by vesamicol and omega-agatoxin IVA.
Stimulation of mammalian motor neurons can elicit Ca(2+)-dependent regenerative release of acetylcholine and prolonged endplate depolarization when the enzymatic degradation of the neurotransmitter is inhibited. Unlike physiological phasic release of acetylcholine, the regenerative release is sensitive to L-type Ca2+ channel blockers. We studied the effects of vesamicol (an inhibitor of active transport of acetylcholine into synaptic vesicles) and omega-agatoxin IVA (a blocker of the motor nerve P-type Ca2+ channel) on these two types of acetylcholine release to compare the vesicle pools and Ca2+ channels responsible for the release. When coupled with repetitive stimulations, vesamicol decreased mean amplitude of miniature endplate potentials, resulting in a skewed distribution to lower amplitude, reduced quantal content of endplate potentials and decreased immediate available pool of acetylcholine. omega-Agatoxin IVA had no effect on miniature endplate potential but inhibited quantal content of endplate potential. The mean inhibitory concentration was around 5-10 nM. Vesamicol and omega-agatoxin IVA decreased the probability of triggering regenerative release. However, the magnitude and duration of regenerative release, once triggered, were not depressed by either agent. It appears that the majority of Ca2+ necessary for regenerative release is translocated via omega-agatoxin IVA-insensitive Ca2+ channels, which can be activated by prolonged depolarization of nerve terminals induced by accumulated acetylcholine. The results suggest that different Ca2+ channels are activated in the regenerative (L-type) and phasic (P-type) acetylcholine release, which utilize different pools of synaptic vesicles.
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D003964:Diaphragm / Q000187:drug effects / Q000294:innervation; D004558:Electric Stimulation; D005071:Evoked Potentials / Q000187:drug effects; D066298:In Vitro Techniques; D051379:Mice; D008813:Mice, Inbred ICR; D009045:Motor Endplate / Q000187:drug effects; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism*; D009388:Neostigmine / Q000494:pharmacology; D009416:Nerve Regeneration / Q000187:drug effects / Q000502:physiology*; D009467:Neuromuscular Depolarizing Agents / Q000494:pharmacology*; D010791:Phrenic Nerve / Q000187:drug effects; D010880:Piperidines / Q000494:pharmacology*; D013111:Spider Venoms / Q000494:pharmacology*; D020867:omega-Agatoxin IVA
D002121:Calcium Channel Blockers; D009467:Neuromuscular Depolarizing Agents; D010880:Piperidines; D013111:Spider Venoms; D020867:omega-Agatoxin IVA; D009388:Neostigmine; C006189:vesamicol; D000109:Acetylcholine