Dual regulation by protein kinase C of the muscarinic response in Xenopus oocytes.

Muscarinic stimulation of follicle-enclosed oocytes of Xenopus laevis results in a complex response that involves both depolarizing and hyperpolarizing currents (Dascal and Landau 1980). We studied the involvement of protein kinase C (PK-C1) in the regulation of the acetylcholine-evoked rapid (D1) and of the slow (D2) depolarizing chloride (Cl-) currents. In oocytes maintained at -100 mV [the reversal potential of potassium (K+) ions] under two electrode voltage clamp, the PK-C activatory 4-beta-phorbol 12-myristate 13-acetate (beta-PMA, 0.1 microM) stimulated D1 by 99 +/- 17% and inhibited D2 by 67 +/- 6%, vs. untreated controls. The inactive isomer (alpha-PMA) or phorbol alone had no significant effect on the components of the muscarinic response. In order to identify the site of the regulation, we have microinjected the intracellular second messenger of calcium mobilization, inositol 1,4,5-trisphosphate (IP3). beta-PMA or the diacylglycerol analog, oleoylacetylglycerol (OAG) stimulated the rapid depolarizing current evoked by IP3 by 220 +/- 26% and 394 +/- 102%, respectively. alpha-PMA had little if any effect. The calcium-evoked Cl- current in oocytes pre-treated with the divalent cation ionophore A23187 was, on the other hand, inhibited by beta-PMA and OAG (by 82 +/- 6% and 54 +/- 6%, respectively). alpha-PMA and phorbol had a limited inhibitory effect. beta-PMA, but not alpha-PMA, also mildly inhibited the IP3-evoked increase in 45Ca efflux. The intracellular metabolism of IP3 was not affected by exposure to either beta-PMA or OAG.(ABSTRACT TRUNCATED AT 250 WORDS)