Nicotinic receptor-evoked release of acetylcholine and somatostatin in the myenteric plexus is coupled to calcium influx via N-type calcium channels.
Entry of extracellular calcium (Ca++) via voltage-gated Ca++ channels is essential for neurotransmitter release. In this study, we examined whether nicotinic receptor-stimulated release of acetylcholine (ACh) and somatostatin (S14) are coupled to calcium influx via distinct calcium channel subtypes in the myenteric plexus. Isolated ganglia from the guinea pig ileal myenteric plexus were prepared and placed in perfusion chambers under standard conditions. The ganglionic agonist dimethylphenylpiperazinium (DMPP, 10(-6) to 10(-3) M) stimulated the release of [3H]ACh in a concentration-dependent manner. This release was blocked by hexamethonium or Ca(++)-free medium containing 1 mM EGTA and was antagonized by omega-conotoxin, a preferential N calcium channel blocker, but was not affected by nifedipine (L channel antagonist) or nickel (T calcium channel antagonist). DMPP-evoked release of somatostatin was also antagonized by omega-conotoxin, but was not affected by nifedipine or nickel. These observations indicate that neurosecretion of ACh and S14 evoked by DMPP is mediated by calcium entry via voltage-sensitive N-type Ca++ channels. To provide additional evidence that nicotinic receptor stimulation is associated with Ca++ entry via the N-type Ca++ channels, we examined the intracellular calcium [Ca++]i concentration of the myenteric plexus neurons using fura-2 microspectrofluorometry. Basal [Ca++]i of single ileal myenteric neurons was 65 +/- 5 nM. Perfusion with DMPP (10(-6) to 10(-3) M) caused a rapid, transient elevation in [Ca++]i which was abolished by Ca(++)-free medium containing 1 mM EGTA.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor.
T
T
Takahashi
Y
Y
Tsunoda
Y
Y
Lu
J
J
Wiley
C
C
Owyang
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D006168:Guinea Pigs; D008297:Male; D009197:Myenteric Plexus / Q000187:drug effects* / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000502:physiology; D013004:Somatostatin / Q000378:metabolism*
D002120:Calcium Channel Agonists; D002121:Calcium Channel Blockers; D011978:Receptors, Nicotinic; D013004:Somatostatin; D000109:Acetylcholine; D002118:Calcium