Modulation of luteinizing hormone-stimulated inositol phosphate accumulation by phorbol esters in bovine luteal cells

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Abstract

The present studies were conducted to evaluate the effects of protein kinase C activators on the inositol phospholipid-phospholipase C second messenger system in isolated bovine luteal cells. This report describes the effects of phorbol esters on inositol phosphate accumu¬lation in LH- and prostaglandin F2α (PGF2α)-stimulated bovine luteal cells. Corpora lutea of early pregnancy were dispersed with collagenase and luteal cells were prelabeled for 3 h with [3H]inositol. Inositol phosphates produced in response to LH or PGF2„ were analyzed by ion exchange column chromatography. The tumor promoter and protein kinase C activator 12-O-tetrade-canolyphorbol 13-acetate (TPA) had no effect on basal levels of inositol phosphates but inhibited LH-stimulated accumulation of inositol mono-, bis-, and trisphosphates by 72%, 68%, and 65%, respectively. TPA reduced the response to maximally effective concentrations of LH and tripled the concentrations of LH required to evoke half- maximal accumulation of inositol mono-, bis-. risphosphates. The inhibitory effects of TPA were rapid (5 min) whether added before or after treatment with LH. Treatment with TPA also reduced (58%) the initial phase of intracellular calcium mobilization in LH-treated cells The inhibitory effects of TPA were not associated with acute reductions in [3H]inositol incorporation, [3H]inositol phospholipid levels, cAMP levels, or progesterone accumulation in control or LH-stimulated luteal cells. The effects of phorbol esters were concentration dependent and specific for active tumor promoters with 10-50 nM TPA producing maximal inhibitory effects. A synthetic diacylglycerol, l-oleyl-2-acetyl- glycerol, mimicked the inhibitory effects of TPA. In contrast, pretreatment with a physiological activator of protein kinase C, PGF2α, had no effect on LH-stimulated inositol phosphate accumulation. The inhibi¬tory effects of TPA could not be explained by a generalized inhibition of phospholipase C or G-proteins since the accumulation of inositol phosphates in PGF2α-and NaF-treated cells was not inhibited by TPA. These results demonstrate that tumor promoting phorbol esters modulate the inositol phospholipid-phospholipase C transmembrane signaling system in LH-stimulated bovine luteal cells. The results suggest that phorbol esters may alter the coupling of the LH-receptor complex to phospholipase C. These findings implicate protein kinase C in the regulation of transmembrane signaling in the bovine corpus luteum.

Original languageEnglish (US)
Pages (from-to)749-757
Number of pages9
JournalEndocrinology
Volume131
Issue number2
DOIs
StatePublished - Aug 1992

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Luteal Cells
Inositol Phosphates
Phorbol Esters
Luteinizing Hormone
Dinoprost
Type C Phospholipases
Inositol
Protein Kinase C
Phosphatidylinositols
Corpus Luteum
Carcinogens
Gonadotrophs
LH Receptors
Diglycerides
Ion Exchange Chromatography
Second Messenger Systems
Collagenases
GTP-Binding Proteins
Progesterone
Acetates

ASJC Scopus subject areas

  • Endocrinology

Cite this

Modulation of luteinizing hormone-stimulated inositol phosphate accumulation by phorbol esters in bovine luteal cells. / Davis, John S.

In: Endocrinology, Vol. 131, No. 2, 08.1992, p. 749-757.

Research output: Contribution to journalArticle

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abstract = "The present studies were conducted to evaluate the effects of protein kinase C activators on the inositol phospholipid-phospholipase C second messenger system in isolated bovine luteal cells. This report describes the effects of phorbol esters on inositol phosphate accumu¬lation in LH- and prostaglandin F2α (PGF2α)-stimulated bovine luteal cells. Corpora lutea of early pregnancy were dispersed with collagenase and luteal cells were prelabeled for 3 h with [3H]inositol. Inositol phosphates produced in response to LH or PGF2„ were analyzed by ion exchange column chromatography. The tumor promoter and protein kinase C activator 12-O-tetrade-canolyphorbol 13-acetate (TPA) had no effect on basal levels of inositol phosphates but inhibited LH-stimulated accumulation of inositol mono-, bis-, and trisphosphates by 72{\%}, 68{\%}, and 65{\%}, respectively. TPA reduced the response to maximally effective concentrations of LH and tripled the concentrations of LH required to evoke half- maximal accumulation of inositol mono-, bis-. risphosphates. The inhibitory effects of TPA were rapid (5 min) whether added before or after treatment with LH. Treatment with TPA also reduced (58{\%}) the initial phase of intracellular calcium mobilization in LH-treated cells The inhibitory effects of TPA were not associated with acute reductions in [3H]inositol incorporation, [3H]inositol phospholipid levels, cAMP levels, or progesterone accumulation in control or LH-stimulated luteal cells. The effects of phorbol esters were concentration dependent and specific for active tumor promoters with 10-50 nM TPA producing maximal inhibitory effects. A synthetic diacylglycerol, l-oleyl-2-acetyl- glycerol, mimicked the inhibitory effects of TPA. In contrast, pretreatment with a physiological activator of protein kinase C, PGF2α, had no effect on LH-stimulated inositol phosphate accumulation. The inhibi¬tory effects of TPA could not be explained by a generalized inhibition of phospholipase C or G-proteins since the accumulation of inositol phosphates in PGF2α-and NaF-treated cells was not inhibited by TPA. These results demonstrate that tumor promoting phorbol esters modulate the inositol phospholipid-phospholipase C transmembrane signaling system in LH-stimulated bovine luteal cells. The results suggest that phorbol esters may alter the coupling of the LH-receptor complex to phospholipase C. These findings implicate protein kinase C in the regulation of transmembrane signaling in the bovine corpus luteum.",
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