Evidence for altered circular smooth muscle cell function in lower esophageal sphincter ofW/Wvmutant mice

Abstract

Nitrergic neurotransmission to gut smooth muscle is impaired in W/Wvmutant mice, which lack intramuscular interstitial cells of Cajal (ICC-IM). In addition, these mice have been reported to have smaller amplitude unitary potentials (UPs) and a more negative resting membrane potential (RMP) than control mice. These abnormalities have been attributed to absence of ICC-IM, but it remains possible that they are due to alterations at the level of the smooth muscle itself. Amphotericin-B-perforated patch-clamp recordings and Ca2+imaging (fura 2) were compared between freshly isolated single circular smooth muscle cells (CSM) from W/Wvmutant and control mice lower esophageal sphincter (LES). There was no significant difference in seal resistance, capacitance, or input resistance in response to applied electrotonic current pulses between CSM cells from W/Wvmutants and controls. Compared with control mice, RMP was more negative and UPs significantly smaller in CSM cells from mutant mice LES. Administration of caffeine induced an inward current in cells from both mutant and control mice, but the current density was significantly larger in cells from W/Wvmutants. Membrane potential hyperpolarization induced by sodium nitroprusside was larger in cells from control mice vs. W/Wvmutants. In addition, intracellular Ca2+transients induced by caffeine were significantly increased in cells from mutants. These findings indicate that LES CSM is abnormal in W/Wvmutant mice. Thus some physiological functions attributed to ICC-IM based on experiments in smooth muscle of ICC deficient mice may need to be reconsidered.