Dopamine inhibits basal prolactin release in pituitary lactotrophs through pertussis toxin-sensitive and insensitive signaling pathways

Dopamine D2 receptors signal through the pertussis toxin (PTX)-sensitiveG(i/o) and PTX-insensitive G(z) proteins, as well as through a G protein-independent, beta-arrestin/glycogen synthase kinase-3-dependent pathway. Activation of these receptors in pituitary lactotrophs leads to inhibition of prolactin (PRL) release. It has been suggested that this inhibition occurs through the G(i/o)-alpha protein-mediated inhibition of cAMP production and/or G(i/o)-beta gamma dimer-mediated activation of inward rectifier K+ channels and inhibition of voltage-gated Ca2+ channels. Here we show that the dopamine agonist-induced inhibition of spontaneous Ca2+ influx and release of prestored PRL was preserved when cAMP levels were elevated by forskolin treatment. We further observed that dopamine agonists inhibited both spontaneous and depolarizationinduced Ca2+ influx in untreated but not in PTX-treated cells. This inhibition was also observed in cells with blocked inward rectifier K+ channels, suggesting that the dopamine effect on voltage-gated Ca2+ channel gating is sufficient to inhibit spontaneous Ca2+ influx. However, agonist-induced inhibition of PRL release was only partially relieved in PTX-treated cells, indicating that dopamine receptors also inhibit exocytosis downstream of voltage-gated Ca2+ influx. The PTX-insensitive step in agonist-induced inhibition of PRL release was not affected by the addition of wortmannin, an inhibitor of phosphatidylinositol 3-kinase, and lithium, an inhibitor of glycogen synthase kinase-3, but was attenuated in the presence of phorbol 12-myristate 13-acetate, which inhibits G(z) signaling pathway in a protein kinase C-dependent manner. Thus, dopamine inhibits basal PRL release by blocking voltage-gated Ca2+ influx through the PTX-sensitive signaling pathway and by desensitizing Ca2+ secretion coupling through the PTX-insensitive and protein kinase C-sensitive signaling pathway.