Phorbol ester-induced M-current suppression in bull-frog sympathetic ganglion cells: insensitivity to kinase inhibitors.

1. The effects of 1-oleoyl-2-acetyl-sn-glycerol (OAG), phorbol 12-myristate 13-acetate (PMA), 4-alpha-phorbol and muscarine on B-neurones from bull-frog sympathetic ganglion were studied by means of whole-cell patch-clamp recording. With the exception of 4-alpha-phorbol, all of these agonists reduced the steady-state outward current recorded at -30 mV as a result of suppression of a voltage-dependent, non-inactivating K(+)-current, the M-current, (IM). 2. Of the cells tested, 34% displayed bona fide responses to OAG (20 microM). The chance of recording a response was not decreased when the protein kinase inhibitor, 1-(5-isoquinolinylsulphonyl)-2-methyl-piperazine (H-7; 50 or 75 microM) was included simultaneously in the extracellular solution and in the pipette solution. 3. The presence of 50 microM H-7 on both sides of the membrane or 500 nM staurosporine in the pipette solution did not prevent responses to brief (1-2 min) or prolonged (> 20 min) applications of PMA. 4. Brief (1-2 min) extracellular application of H-7 (300 microM) suppressed IM by about 29%. 5. The most likely explanation of these data is that PMA and OAG modulate IM via a mechanism that is independent of protein kinase C (PKC). The availability of such a mechanism poses new questions as to the mechanism of muscarine-induced IM suppression.