Müllerian inhibiting substance blocks the protein kinase A-induced expression of cytochrome P450 17α-hydroxylase/C17-20 lyase mRNA in a mouse Leydig cell line independent of cAMP responsive element binding protein phosphorylation

V. Matt Laurich, Alexander M. Trbovich, Francis H. O'Neill, Christopher P. Houk, Patrick M. Sluss, Anita H. Payne, Patricia K. Donahoe, Jose Teixeira

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Müllerian inhibiting substance (MIS) is produced by fetal Sertoli cells and causes regression of the Müllerian duct in male fetuses shortly after commitment of the bipotential embryonic gonad to testes differentiation. MIS is also produced by the Sertoli cells and granulosa cells of the adult gonads where it plays an important role in regulating steroidogenesis. We have previously shown that MIS can dramatically reduce testosterone synthesis in Leydig cells by inhibiting the expression of cytochrome P450 17α-hydroxylase/C17-20 lyase (Cyp17) mRNA in vitro and in vivo. To characterize the signal transduction pathway used by MIS to control expression of endogenous Cyp17 in a mouse Leydig cell line, we demonstrate that MIS inhibits both LH- and cAMP-induced expression of Cyp17 at concentrations as low as 3.5 nM and for as long as 18 h. The induction of steroidogenic acute regulatory protein (STAR) mRNA by cAMP, however, was slightly increased by addition of MIS. Protein kinase A (PKA) inhibition with H-89 blocked Cyp17 mRNA induction, suggesting that MIS interferes with the PKA signal transduction pathway. Inhibition of Cyp17 induction was not seen with added U0126, and wortmannin inhibited the induction incompletely. In addition, phosphorylation of the cAMP responsive element binding protein (CREB) was not detected following 50 μM cAMP exposure, a concentration sufficient for Cyp17 mRNA induction. Moreover, CREB phosphorylation, which was observed with addition of 500 μM cAMP, was not inhibited by coincubation with MIS. Taken together, these results suggest that cAMP induces expression of Cyp17 by a PKA-mediated mechanism and that this induction, which is inhibited by MIS signal transduction, does not require CREB activity, and is distinct from that used to induce steroidogenic acute regulatory protein expression.

Original languageEnglish (US)
Pages (from-to)3351-3360
Number of pages10
JournalEndocrinology
Volume143
Issue number9
DOIs
StatePublished - Sep 1 2002
Externally publishedYes

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Lyases
Leydig Cells
Mixed Function Oxygenases
Cyclic AMP-Dependent Protein Kinases
Cytochrome P-450 Enzyme System
Carrier Proteins
Phosphorylation
Cell Line
Signal Transduction
Messenger RNA
Sertoli Cells
Gonads
Granulosa Cells
Testosterone
Testis
Fetus
steroidogenic acute regulatory protein

ASJC Scopus subject areas

  • Endocrinology

Cite this

Müllerian inhibiting substance blocks the protein kinase A-induced expression of cytochrome P450 17α-hydroxylase/C17-20 lyase mRNA in a mouse Leydig cell line independent of cAMP responsive element binding protein phosphorylation. / Matt Laurich, V.; Trbovich, Alexander M.; O'Neill, Francis H.; Houk, Christopher P.; Sluss, Patrick M.; Payne, Anita H.; Donahoe, Patricia K.; Teixeira, Jose.

In: Endocrinology, Vol. 143, No. 9, 01.09.2002, p. 3351-3360.

Research output: Contribution to journalArticle

Matt Laurich, V. ; Trbovich, Alexander M. ; O'Neill, Francis H. ; Houk, Christopher P. ; Sluss, Patrick M. ; Payne, Anita H. ; Donahoe, Patricia K. ; Teixeira, Jose. / Müllerian inhibiting substance blocks the protein kinase A-induced expression of cytochrome P450 17α-hydroxylase/C17-20 lyase mRNA in a mouse Leydig cell line independent of cAMP responsive element binding protein phosphorylation. In: Endocrinology. 2002 ; Vol. 143, No. 9. pp. 3351-3360.
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abstract = "M{\"u}llerian inhibiting substance (MIS) is produced by fetal Sertoli cells and causes regression of the M{\"u}llerian duct in male fetuses shortly after commitment of the bipotential embryonic gonad to testes differentiation. MIS is also produced by the Sertoli cells and granulosa cells of the adult gonads where it plays an important role in regulating steroidogenesis. We have previously shown that MIS can dramatically reduce testosterone synthesis in Leydig cells by inhibiting the expression of cytochrome P450 17α-hydroxylase/C17-20 lyase (Cyp17) mRNA in vitro and in vivo. To characterize the signal transduction pathway used by MIS to control expression of endogenous Cyp17 in a mouse Leydig cell line, we demonstrate that MIS inhibits both LH- and cAMP-induced expression of Cyp17 at concentrations as low as 3.5 nM and for as long as 18 h. The induction of steroidogenic acute regulatory protein (STAR) mRNA by cAMP, however, was slightly increased by addition of MIS. Protein kinase A (PKA) inhibition with H-89 blocked Cyp17 mRNA induction, suggesting that MIS interferes with the PKA signal transduction pathway. Inhibition of Cyp17 induction was not seen with added U0126, and wortmannin inhibited the induction incompletely. In addition, phosphorylation of the cAMP responsive element binding protein (CREB) was not detected following 50 μM cAMP exposure, a concentration sufficient for Cyp17 mRNA induction. Moreover, CREB phosphorylation, which was observed with addition of 500 μM cAMP, was not inhibited by coincubation with MIS. Taken together, these results suggest that cAMP induces expression of Cyp17 by a PKA-mediated mechanism and that this induction, which is inhibited by MIS signal transduction, does not require CREB activity, and is distinct from that used to induce steroidogenic acute regulatory protein expression.",
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AU - Matt Laurich, V.

AU - Trbovich, Alexander M.

AU - O'Neill, Francis H.

AU - Houk, Christopher P.

AU - Sluss, Patrick M.

AU - Payne, Anita H.

AU - Donahoe, Patricia K.

AU - Teixeira, Jose

PY - 2002/9/1

Y1 - 2002/9/1

N2 - Müllerian inhibiting substance (MIS) is produced by fetal Sertoli cells and causes regression of the Müllerian duct in male fetuses shortly after commitment of the bipotential embryonic gonad to testes differentiation. MIS is also produced by the Sertoli cells and granulosa cells of the adult gonads where it plays an important role in regulating steroidogenesis. We have previously shown that MIS can dramatically reduce testosterone synthesis in Leydig cells by inhibiting the expression of cytochrome P450 17α-hydroxylase/C17-20 lyase (Cyp17) mRNA in vitro and in vivo. To characterize the signal transduction pathway used by MIS to control expression of endogenous Cyp17 in a mouse Leydig cell line, we demonstrate that MIS inhibits both LH- and cAMP-induced expression of Cyp17 at concentrations as low as 3.5 nM and for as long as 18 h. The induction of steroidogenic acute regulatory protein (STAR) mRNA by cAMP, however, was slightly increased by addition of MIS. Protein kinase A (PKA) inhibition with H-89 blocked Cyp17 mRNA induction, suggesting that MIS interferes with the PKA signal transduction pathway. Inhibition of Cyp17 induction was not seen with added U0126, and wortmannin inhibited the induction incompletely. In addition, phosphorylation of the cAMP responsive element binding protein (CREB) was not detected following 50 μM cAMP exposure, a concentration sufficient for Cyp17 mRNA induction. Moreover, CREB phosphorylation, which was observed with addition of 500 μM cAMP, was not inhibited by coincubation with MIS. Taken together, these results suggest that cAMP induces expression of Cyp17 by a PKA-mediated mechanism and that this induction, which is inhibited by MIS signal transduction, does not require CREB activity, and is distinct from that used to induce steroidogenic acute regulatory protein expression.

AB - Müllerian inhibiting substance (MIS) is produced by fetal Sertoli cells and causes regression of the Müllerian duct in male fetuses shortly after commitment of the bipotential embryonic gonad to testes differentiation. MIS is also produced by the Sertoli cells and granulosa cells of the adult gonads where it plays an important role in regulating steroidogenesis. We have previously shown that MIS can dramatically reduce testosterone synthesis in Leydig cells by inhibiting the expression of cytochrome P450 17α-hydroxylase/C17-20 lyase (Cyp17) mRNA in vitro and in vivo. To characterize the signal transduction pathway used by MIS to control expression of endogenous Cyp17 in a mouse Leydig cell line, we demonstrate that MIS inhibits both LH- and cAMP-induced expression of Cyp17 at concentrations as low as 3.5 nM and for as long as 18 h. The induction of steroidogenic acute regulatory protein (STAR) mRNA by cAMP, however, was slightly increased by addition of MIS. Protein kinase A (PKA) inhibition with H-89 blocked Cyp17 mRNA induction, suggesting that MIS interferes with the PKA signal transduction pathway. Inhibition of Cyp17 induction was not seen with added U0126, and wortmannin inhibited the induction incompletely. In addition, phosphorylation of the cAMP responsive element binding protein (CREB) was not detected following 50 μM cAMP exposure, a concentration sufficient for Cyp17 mRNA induction. Moreover, CREB phosphorylation, which was observed with addition of 500 μM cAMP, was not inhibited by coincubation with MIS. Taken together, these results suggest that cAMP induces expression of Cyp17 by a PKA-mediated mechanism and that this induction, which is inhibited by MIS signal transduction, does not require CREB activity, and is distinct from that used to induce steroidogenic acute regulatory protein expression.

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