M-current suppression by agonist and phorbol ester in bullfrog sympathetic neurons

Abstract

Activation of protein kinase C (PKC) by phorbol esters is known to suppress M-current. 4-beta-Phorbol 12,13-dibutyrate (PDBu) irreversibly suppressed M-current in a concentration-dependent manner (Ki 38 nM). Inhibitors of PKC, the pseudo-substrate peptide PKCI (19-31), staurosporine and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H7) antagonized PDBu-mediated suppression of M-current. Suppression of M-current by muscarine and luteinizing hormone-releasing hormone (LHRH) was unaffected by PKCI (19-31) and H7, but was antagonized by staurosporine. The balance of data suggests that suppression of M-current by agonists is probably not mediated by activation of PKC. Addition and subsequent removal of PDBu to M-current suppressed by muscarine prevented the action of PDBu, while closing M-channels by voltage or blocking by barium did not. This suggests that M-channel closure by muscarine protects those channels from the effects of PDBu. Partial suppression of M-current by low concentrations of muscarine antagonized the response to PDBu, with the magnitude of suppression equivalent to that seen with PDBu alone. It is suggested that two interconvertable populations of M-channels exist, one that is sensitive to both agonist and PDBu and another that can only be suppressed by agonist.