nitroarginine and 2-aminoethoxydiphenyl-borate

We studied pacing and neurotransmission in longitudinal (LM) and circular muscle (CM) in intestine of W/W++ and W/W(V) mice. Electrical field-stimulation (EFS) of nerves in LM segments was more inhibitory in W/W(V) mice than in W/W++ mice. No inhibitory input to CM segments of W/W(V) mice was found. The EFS, after nerve block, entrained segments of both W/W++ and mutant mice with 10 ms pulses, and entrained those of mutant mice more readily at 1 and 3 ms pulses. Pacing with external electrodes did not depend on interstitial cells of Cajal in the myenteric plexus (ICC-MP). 2-Aminoethoxydiphenyl borate (2-APB), putative antagonist at IP3 receptors, store-operated channels and the Sacro-endoplasmic reticulum Ca2+ ATPase pump, reduced frequency and amplitudes of pacing of LM segments from W/W(V) mice as it did in BALB/c mice. Thus, its actions may not require ICC-MP. SKF 96365, a putative inhibitor of store-operated channels, reduced frequencies and amplitudes of intestinal segments in W/W++ mice at 10 or 30 micromol L-1. This resulted from blocking L-Ca2+-channels. Thus, no evidence was found that store-operated channels play a role in pacing. In LM segments of W/W(V), SKF 96365 had no effects on frequency of contractions. We conclude, results from models of severely reduced systems may not be applicable to intact ICC networks.

Topics: Anesthetics, Local; Animals; Boron Compounds; Calcium Channel Blockers; Electric Stimulation; Enzyme Inhibitors; Imidazoles; In Vitro Techniques; Intestines; Mice; Mice, Inbred BALB C; Mice, Inbred Strains; Mice, Knockout; Muscle, Smooth; Myenteric Plexus; Nerve Block; Nitroarginine; Proto-Oncogene Proteins c-kit; Species Specificity; Tetrodotoxin

Intracellular recordings were made from isolated bundles of the circular muscle layer of guinea-pig gastric antrum and the responses produced by stimulating intrinsic nerve fibres were examined. After abolishing the effects of stimulating inhibitory nerve terminals with apamin and L-nitroarginine (NOLA), transmural nerve stimulation often evoked a small amplitude excitatory junction potential (EJP) and invariably evoked a regenerative potential. Neurally evoked regenerative potentials had similar properties to those evoked in the same bundle by direct stimulation. EJPs and neurally evoked regenerative potentials were abolished by hyoscine suggesting that both resulted from the release of acetylcholine and activation of muscarinic receptors. Neurally evoked regenerative potentials, but not EJPs, were abolished by membrane hyperpolarization, caffeine and chloride channel blockers. In the intact antrum, excitatory vagal nerve stimulation increased the frequency of slow waves. Simultaneous intracellular recordings of pacemaker potentials from myenteric interstitial cells (ICC(MY)) and slow waves showed that the onset of each pacemaker potential normally preceded the onset of each slow wave but vagal stimulation caused the onset of each slow wave to precede each pacemaker potential. Together the observations suggest that during vagal stimulation there is a change in the origin of pacemaker activity with slow waves being initiated by intramuscular interstitial cells (ICC(IM)) rather than by ICC(MY).

Topics: 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; Animals; Apamin; Biological Clocks; Boron Compounds; Electric Stimulation; Evoked Potentials; Excitatory Postsynaptic Potentials; Female; Guinea Pigs; In Vitro Techniques; Male; Membrane Potentials; Muscarinic Antagonists; Muscle, Smooth; Nerve Endings; Nitroarginine; Pyloric Antrum; Scopolamine; Stomach; Vagus Nerve