Muscular effects of orexin A on the mouse duodenum: mechanical and electrophysiological studies

Orexin A (OXA) has been reported to influence gastrointestinal motility, acting at both central and peripheral neural levels. The aim of the present study was to evaluate whether OXA also exerts direct effects on the duodenal smooth muscle. The possible mechanism of action involved was investigated by employing a combined mechanical and electrophysiological approach. Duodenal segments were mounted in organ baths for isometric recording of the mechanical activity. Ionic channel activity was recorded in current-and voltage-clamp conditions by a single microelectrode inserted in a duodenal longitudinal muscle cell. In the duodenal preparations, OXA(0.3 mu M) caused a TTX-insensitive transient contraction. Nifedipine (1 mu M), as well as 2-aminoethyl diphenyl borate (10 mu M), reduced the amplitude and shortened the duration of the response to OXA, which was abolished by Ni2+ (50 mu M) or TEA(1 mM). Electrophysiological studies in current-clamp conditions showed that OXA caused an early depolarization, which paralleled in time the contractile response, followed by a long-lasting depolarization. Such a depolarization was triggered by activation of receptor-operated Ca2+ channels and enhanced by activation of T-and L-type Ca2+ channels and store-operated Ca2+ channels and by inhibition of K+ channels. Experiments in voltage-clamp conditions demonstrated that OXA affects not only receptor-operatedCa(2+) channels, but also the maximal conductance and kinetics of activation and inactivation of Na+, T-and L-typeCa(2+) voltage-gated channels. The results demonstrate, for the first time, that OXA exerts direct excitatory effects on the mouse duodenal smooth muscle. Finally, this work demonstrates new findings related to the expression and kinetics of the voltage-gated channel types, as well as store-operated Ca2+ channels.