Activation and inhibition of protein kinase C in cultured bovine parathyroid cells: effect on the release of C-terminal fragments of parathyroid hormone.
The response of the parathyroid gland to low Ca2+ may be mediated in part by protein kinase C (PKC). We assessed the effect of two PKC activators, SC-9 and SC-10, and one PKC inhibitor, H-7, on Ca(2+)-regulated PTH release and degradation in primary cultures of bovine parathyroid cells. Both SC-9 and SC-10 stimulated PTH release, compared to high Ca2+ alone, in parathyroid cells incubated in high Ca2+, with maximal PTH release of at least twofold occurring at a concentration of either activator of 10 nM (p less than 0.05). We have previously shown that another PKC activator, PMA, not only enhances PTH release in the presence of high Ca2+ but suppresses low Ca(2+)-stimulated PTH secretion. In the present study, neither SC-9 nor SC-10 caused a comparable suppression of PTH release at low Ca2+. However, the PKC inhibitor, H-7 (1 microM), blocked low Ca(2+)-stimulated (compared to the low Ca2+ control) PTH secretion by approximately 50% (p less than 0.01) and did not affect high Ca2+ suppression of PTH secretion. H-7 (1 microM) was able to oppose the stimulation of PTH release by the PKC activators SC-9, SC-10, and PMA at high Ca2+ and negated the PTH release-inhibiting effect of PMA at low Ca2+. Culture medium from these experiments was subjected to reversed-phase HPLC and the eluted fractions analyzed by RIA for the presence of intact and C-terminal fragments of PTH.(ABSTRACT TRUNCATED AT 250 WORDS)
Watson
P H
PH
Department of Medicine, Foothills Hospital, Alberta, Canada.
Mortimer
S T
ST
Tanguay
K E
KE
Hanley
D A
DA
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000818:Animals; D002118:Calcium / Q000502:physiology*; D002417:Cattle; D004338:Drug Combinations; D004789:Enzyme Activation / Q000187:drug effects; D007546:Isoquinolines / Q000494:pharmacology*; D009281:Naphthalenes / Q000494:pharmacology*; D010281:Parathyroid Hormone / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D010879:Piperazines / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D013268:Stimulation, Chemical; D013449:Sulfonamides / Q000494:pharmacology*
D004338:Drug Combinations; D007546:Isoquinolines; D009281:Naphthalenes; D010281:Parathyroid Hormone; D010446:Peptide Fragments; D010879:Piperazines; D013449:Sulfonamides; C049999:N-(6-phenylhexyl)-5-chloro-1-naphthalenesulfonamide; C077158:N-(m-heptyl)-5-chloro-1-naphthalenesulfonamide; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D002118:Calcium