title: Muscular effects of orexin A on the mouse duodenum: mechanical and electrophysiological studies.
abstract: 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 μM) caused a TTX-insensitive transient contraction. Nifedipine (1 μM), as well as 2-aminoethyl diphenyl borate (10 μM), reduced the amplitude and shortened the duration of the response to OXA, which was abolished by Ni(2+) (50 μM) 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 Ca(2+) channels and enhanced by activation of T- and L-type Ca(2+) channels and store-operated Ca(2+) channels and by inhibition of K(+) channels. Experiments in voltage-clamp conditions demonstrated that OXA affects not only receptor-operated Ca(2+) channels, but also the maximal conductance and kinetics of activation and inactivation of Na(+), T- and L-type Ca(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 Ca(2+) channels.
authors.affiliation: Dipartimento di Scienze Fisiologiche, Università di Firenze, Firenze, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Squecco
authors.forename: Rachele
authors.initials: R
authors.lastname: Garella
authors.forename: Giorgia
authors.initials: G
authors.lastname: Luciani
authors.forename: Fabio
authors.initials: F
authors.lastname: Francini
authors.forename: Maria Caterina
authors.initials: MC
authors.lastname: Baccari
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D004386:Duodenum / Q000187:drug effects* / Q000502:physiology; D005260:Female; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins / Q000494:pharmacology*; D007537:Isometric Contraction / Q000187:drug effects; D051379:Mice; D009130:Muscle, Smooth / Q000187:drug effects* / Q000502:physiology; D009479:Neuropeptides / Q000494:pharmacology*; D009543:Nifedipine / Q000494:pharmacology; D000068797:Orexins; D018408:Patch-Clamp Techniques; D026941:Sodium Channel Blockers / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology
chemical_list: C538758:2-aminoethyl diphenylborinate; D001896:Boron Compounds; D002121:Calcium Channel Blockers; D047908:Intracellular Signaling Peptides and Proteins; D009479:Neuropeptides; D000068797:Orexins; D026941:Sodium Channel Blockers; D013779:Tetrodotoxin; D009543:Nifedipine