Regulation of L-alanine transport systems A and ASC by cyclic AMP and calcium in a reptilian duodenal model

Abstract

SUMMARYThe regulation of neutral amino acid transport by cyclic AMP (cAMP) and calcium across the isolated duodenum of the lizard Gallotia gallotihas been studied under short-circuit conditions. Active L-alanine transport was stimulated by forskolin, theophylline and dibutyryl cyclic AMP (db-cAMP). All these agents increased transmural potential difference (PD) and short-circuit current (Isc) in a manner consistent with the activation of a chloride secretory pathway. Both forskolin and theophylline increased intracellular cAMP levels in the lizard duodenal mucosa. Addition of calcium ionophore A23187 rapidly reduced mucosa-to-serosa L-alanine fluxes and diminished net L-alanine transport. Despite the reduction of alanine fluxes by A23187, transepithelial PD and Iscvalues were increased by the ionophore. Analyses of the responses of isolated transport pathways indicated that the Na+-independent L-alanine transport system was unaffected by db-cAMP or calcium ionophore. By contrast,Na+-dependent transport activities were profoundly modified by these agents. Thus, while system A [α-methylamino-isobutiric acid(MeAIB)-transporting pathway] was stimulated by increased calcium, system ASC activity was nearly abolished. Calcium ionophore also potentiated the electrogenic response of system A. Forskolin strongly stimulated system ASC activity but left system A activity unchanged. Activation of system ASC by forskolin was clearly electroneutral, as pre-incubation of the tissues with the chloride channel blocker diphenylamine-2-carboxilic acid (DPC) completely prevented forskolin-induced transepithelial electrical responses. It is concluded that intracellular messengers cAMP and calcium oppositely modulate active Na+-dependent L-alanine transport in the lizard intestine. The different sensitivity exhibited by individual transport pathways may well account for the changes observed in overall alanine transport.