Detection of nitric oxide release from canine enteric neurons.
Previous pharmacological and immunohistochemical studies have suggested that nitric oxide (NO) is a mediator of enteric neuromuscular transmission in the canine proximal colon. The present study demonstrates the release of nitric oxide (NO) and/or its oxidation products (collectively termed NOx) following stimulation of intrinsic neurons with the nicotinic agonist 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP). Strips of muscularis externa, including the ganglionated myenteric and submucosal plexuses were suspended under tension in modified Krebs solution. DMPP (1-100 microM) produced concentration-dependent inhibition of circular muscle contractile activity and this effect was antagonized by L-nitroarginine methyl ester or L-nitroarginine (100 microM). Tetrodotoxin (TTX, 1 microM) significantly reduced mechanical responses to DMPP (10 microM) but had no effect on responses to high concentrations of DMPP (100 microM). Aliquots of the bathing medium were assayed for NOx after regeneration of NO from NO2- and/or NO3-. NO was measured as chemiluminescence produced by reaction with ozone. Detection was linear over the range 6-25,000 pmol of added NO2- or NO3-. In the absence of drugs, a basal release of 1113.5 +/- 100.4 pmol NOx/g tissue (n = 27) was detected after a 30-min incubation period. DMPP (1-100 microM) stimulated a concentration-dependent increase in NOx to 5099.9 +/- 430 pmol/g per 30 min (n = 17) at 100 microM. NOx release was inhibited by an inhibitor of nitric oxide synthase activity (N(G)-monomethyl-L-arginine) or by reduction in extracellular Ca2+ concentration. DMPP-stimulated NOx accumulation was significantly reduced, but not abolished by TTX (1 microM). These results provide further evidence that NO is released following stimulation of intrinsic neurons in canine proximal colon and support previous suggestions that nicotinic agonists may directly stimulate terminals of NO-releasing neurons.
Shuttleworth
C W
CW
Department of Pharmacology, University of Nevada School of Medicine, Reno 89557, USA.
Weinert
J S
JS
Sanders
K M
KM
Buxton
I L
IL
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D003106:Colon / Q000378:metabolism*; D004246:Dimethylphenylpiperazinium Iodide / Q000494:pharmacology; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D017615:Enteric Nervous System / Q000378:metabolism*; D005260:Female; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects*; D019331:NG-Nitroarginine Methyl Ester; D009569:Nitric Oxide / Q000378:metabolism*; D013779:Tetrodotoxin / Q000494:pharmacology
D009569:Nitric Oxide; D013779:Tetrodotoxin; D004246:Dimethylphenylpiperazinium Iodide; D001120:Arginine; D019331:NG-Nitroarginine Methyl Ester