Muscarinic, cholinergic inputs, largely from the medial septum, have pronounced effects on hippocampal cell excitability. A major effect of synaptically released ACh is block of the slow Ca(2+)-dependent potassium current, called IAHP. Protein kinase C exists in the hippocampus in high concentrations, its activation blocks IAHP, and it has been suggested as a mediator of the muscarinic-receptor-(mAChR)-mediated actions. Using conditions that produce a stable postspike afterhyperpolarizing current (IAHP) in whole-cell recordings from CA1 hippocampal pyramidal neurons in the slice preparation, we have investigated the role of PKC in the cholinergic inhibition of IAHP mediated by mACHRs. Bath application of the general kinase inhibitor, H7, had no effect on inhibition of IAHP by carbachol, although H7 dramatically reduced inhibition of IAHP by the phorbol ester, phorbol-12, 13-diacetate (PDA). Another muscarinic response thought to be mediated by PKC-inhibition of GABAB-mediated hyperpolarization-was reduced by extracellular H7 treatment, suggesting that the coupling between mAChRs and protein kinase activity was maintained in whole-cell recordings. We also discovered that PDA does not mediate its effects on IAHP directly. Intracellular perfusion of high concentrations of H7 (10 mM) or the specific PKC inhibitor, PKCI(19-31) (1 mM), did not prevent inhibition of IAHP by PDA. These results are consistent with an indirect, presynaptic action of phorbol esters on IAHP, possibly mediated through enhanced release of neurotransmitter from surrounding cells.