The differential effects of protein kinase C activators and inhibitors on rat anterior pituitary hormone release.
We investigated the possibility that various protein kinase C (PKC) activators and inhibitors may differentially affect luteinizing hormone (LH) and growth hormone (GH) release from rat anterior pituitary tissue, incubated in vitro. Activators of PKC induced LH release with the following order of potency: mezerein > phorbol 12,13-dibutyrate (PDBu). Mezerein and PDBu were equipotent on GH release. A range of PKC inhibitors (including compounds highly selective for PKC) potently and completely inhibited PKC activator-induced LH and GH release. Chelerythrine and 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H7) were less potent inhibitors of PDBu-induced GH release than of LH release. A component of PDBu- and mezerein-induced LH release was inhibited by H7 with high potency, but a second H7-insensitive component was detected. Mezerein- and PDBu-induced GH release consisted of an H7-resistant component only. When the regulatory domain of PKCs from different sources was investigated by displacement of [3H]PDBu binding, the affinity for mezerein was 3-5-fold greater than that for PDBu at PKCs from cerebral cortex, lung and alpha and beta isoforms extensively purified from brain. Anterior pituitary PKCs were unusual in showing closely matched affinity for mezerein and PDBu, reminiscent of their equivalent potency on GH release. In order to investigate the potency of the catalytic domain inhibitor H7 on PKCs from different sources, enzyme activity assays were carried out on partially purified cytosolic PKCs from midbrain and anterior pituitary and on extensively purified PKC alpha and PKC beta. The Ca(2+)-independent component of PDBu-induced (phosphatidylserine-dependent) activity from anterior pituitary alone showed unusually low potency of inhibition by H7 but was potently inhibited by staurosporine and Ro 31-8220. In contrast, the Ca(2+)-dependent PKC activity in anterior pituitary was inhibited by H7, staurosporine and Ro-31-8220 with high potency as in all other preparations. These results are consistent with the presence and active role in secretion of pharmacologically distinct forms of PKC (or PKC-like kinases) in rat anterior pituitary cells.
Thomson
F J
FJ
MRC Brain Metabolism Unit, University Department of Pharmacology, Edinburgh, UK.
Johnson
M S
MS
Mitchell
R
R
Wolbers
W B
WB
Ison
A J
AJ
MacEwan
D J
DJ
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D053119:Benzophenanthridines; D002273:Carcinogens / Q000494:pharmacology; D002540:Cerebral Cortex / Q000201:enzymology; D003600:Cytosol / Q000378:metabolism; D004224:Diterpenes; D005260:Female; D013006:Growth Hormone / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology*; D007536:Isomerism; D007546:Isoquinolines / Q000494:pharmacology; D008168:Lung / Q000201:enzymology; D007986:Luteinizing Hormone / Q000378:metabolism*; D008301:Maleimides / Q000494:pharmacology; D010617:Phenanthridines / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism / Q000494:pharmacology*; D010879:Piperazines / Q000494:pharmacology; D010903:Pituitary Gland, Anterior / Q000378:metabolism* / Q000473:pathology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D019311:Staurosporine; D013729:Terpenes / Q000378:metabolism / Q000494:pharmacology*; D014316:Tritium
D000470:Alkaloids; D000900:Anti-Bacterial Agents; D053119:Benzophenanthridines; D002273:Carcinogens; D004224:Diterpenes; D007211:Indoles; D007546:Isoquinolines; D008301:Maleimides; D010617:Phenanthridines; D010879:Piperazines; D013729:Terpenes; D014316:Tritium; C011309:mezerein; D015240:Phorbol 12,13-Dibutyrate; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D007986:Luteinizing Hormone; D013006:Growth Hormone; C016299:chelerythrine; D011493:Protein Kinase C; D019311:Staurosporine; C070515:bisindolylmaleimide I; C064758:Ro 31-8220