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Mechanism of action of gonadotropin releasing hormone upon gonadotropin secretion: involvement of protein kinase C as revealed by staurosporine inhibition and enzyme depletion.

Abstract：

:The role of protein kinase C (PKC) in the mechanism of action of gonadotropin-releasing hormone (GnRH) upon gonadotropin secretion is controversial and therefore was investigated in primary cultures of rat anterior pituitary cells. A relatively selective PKC inhibitor, staurosporine, inhibited both GnRH- and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced luteinizing hormone (LH) release with half-maximal inhibition (IC50) of about 80 nM. Inhibition of GnRH action was not complete suggesting also a PKC-insensitive component in GnRH-induced gonadotropin release. Staurosporine had no effect on basal LH release, or on cellular LH content, neither did the drug interfere with the binding of [125I]iodo-[D-Ser(t-Bu)6]des-Gly10-GnRH N-ethylamide to its receptor in pituitary cells. When cultured pituitary cells were incubated with TPA (1 microM) for 24-48 h no measurable cellular PKC activity could be detected. The decrease in total PKC activity was accompanied by an increase in Ca2+, phosphatidylserine (PS), diacylglycerol (DG)-insensitive activity suggesting the release of a portion of the catalytic domain of PKC (M-kinase) by the phorbol ester treatment. TPA-induced LH release was nearly abolished in PKC-depleted cells and the response to GnRH was markedly reduced (40%). The stimulatory effect of the Ca2+ ionophore, ionomycin, was not impaired in PKC-depleted cells. Impaired responses to GnRH in PKC-depleted cells were only noticed at a later phase (2-4 h) of the exocytotic response of the neurohormone. The data strongly suggest a role for PKC during the second phase of GnRH-induced gonadotropin secretion.

journal_name

Mol Cell Endocrinol

journal_title

Molecular and cellular endocrinology

authors

Dan-Cohen H,Naor Z

doi

10.1016/0303-7207(90)90007-u

subject

Has Abstract

pub_date

1990-03-05 00:00:00

pages

135-44

issue

2-3

eissn

0303-7207

issn

1872-8057

journal_volume

69

pub_type

杂志文章

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