Activation of Protein Kinase C by Oxytocin-Related Conopressin Underlies Pacemaker Current in Lymnaea Central Neurons

Abstract

The vasopressin/oxytocin-related neuropeptide Lys-conopressin activates two pacemaker currents in central neurons of the mollusk Lymnaea stagnalis. A high-voltage–activated current ( I-HVA) is activated at potentials greater than −40 mV and resembles pacemaker currents found in many molluscan neurons. A low-voltage–activated current ( I-LVA) activates throughout the range of −90 to 0 mV. Based on sequence homologies, Lymnaea conopressin receptors are thought to couple to Q-type G proteins and protein kinase C (PKC). Alternatively, agonist-induced pacemaker currents in molluscan neurons have traditionally been attributed to cAMP-dependent protein kinase (PKA) activation. Accordingly, this study aimed at resolving possible involvement of cAMP/PKA and diacylglycerol/PKC in the conopressin response. Injection of cAMP into anterior lobe neurons induced a slow inward current with a voltage dependence resembling that of I LVA (and not I HVA). However, lack of effect of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine and the absence of cross-desensitization between cAMP and conopressin suggest that neither current is dependent on intracellular cAMP. The PKC-activating phorbol ester 12- O-tetradecanoylphorbol 13-acetate (but not inactive phorbol 12-myristate 13-acetate) mimicked activation of I HVA, but not I LVA, and occluded subsequent responses to conopressin. Activation of I HVA was blocked by general protein kinase inhibitors and the PKC-inhibitor GF-109203X. Modulation of the calcium buffering capacity of the pipette medium did not affect the conopressin response, suggesting that calcium dynamics are not of major importance. We conclude that conopressin activates the ion channels carrying I LVA and I HVA through different second-messenger cascades and that PKC-dependent phosphorylation underlies activation of I HVA.