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Arginine vasopressin inhibition of cytochrome P450c17 and testosterone production in mouse leydig cells

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Abstract

The mechanism of arginine vasopressin (AVP) action in Leydig cells was investigated, and compared to the effects of phorbol-13-myristate-12-acetate (PMA) and interleukin-1β (IL-1β). Previous reports suggested that AVP inhibits Leydig cell testosterone production at the level of 17α-hydroxylase/C17-lyase activity. The present study confirms and extends these observations, and contrasts the effects of AVP to IL-1. In all experiments, macrophage-depleted Leydig cells were isolated from mice and maintained in primary culture for 5 d prior to initiation of treatments. Leydig cells were treated with 8-Br-cAMP plus increasing concentrations of AVP or IL-1β. AVP caused a significant and dose-dependent inhibition of cAMP-stimulated test-osterone production and P450c17 mRNA expression. IL-1β completely inhibited cAMP-stimulated testosterone production and P450c17 mRNA expression. PMA is a known activator of protein kinase C (PKC) and has been reported to inhibit Leydig cell steroidogenesis. Leydig cells express type V1 vasopressin receptors, which are coupled to PKC activation. The mechanism of IL-1 action in Leydig cells is not understood, but activation of the PKC pathway has been suggested for IL-1 action in other systems. Therefore, the effects of PMA on cAMP-stimulated steroidogenesis were compared to AVP and IL-1. Similar to the effects of AVP, PMA inhibited cAMP-stimulated testosterone production and P450c17 mRNA expression. To assess the possible involvement of PKC in AVP and IL-1 action in Leydig cells, the PKC inhibitor Calphostin C was tested. cAMP-stimulated testosterone production and P450c17 mRNA expression were significantly inhibited by 10 nM AVP (p < 0.05), and this inhibition was reversed by treatment with Calphostin C. Analogous experiments were performed to assess the role of PKC in IL-1 action. In contrast to the results for AVP, Calphostin C did not reverse the inhibitory effects of IL-1 on cAMP-stimulated P450c17 mRNA expression. To assess further PKC activation, myristoylated alanine-rich C kinase substrate (MARCKS) phosphorylation was analyzed. Only AVP and PMA, but not IL-1β, caused an increase in MARCKS phosphorylation. These results confirm that AVP and PMA activate PKC and indicate that IL-1 likely does not activate PKC in Leydig cells. The implications of AVP-mediated inhibition of steroidogenesis and potential role of MARCKS phosphorylation are discussed.

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  1. Department of Physiology and Biophysics (M/C901), University of Illinois at Chicago, 835 S Wolcott Ave., 60612-7342, Chicago, IL

    Dale Buchanan Hales PhD & Robert Greene

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  1. Dale Buchanan Hales PhD
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  2. Robert Greene
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Correspondence to Dale Buchanan Hales PhD.

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Hales, D.B., Greene, R. Arginine vasopressin inhibition of cytochrome P450c17 and testosterone production in mouse leydig cells. Endocr 8, 19–28 (1998). https://doi.org/10.1385/ENDO:8:1:19

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  • DOI: https://doi.org/10.1385/ENDO:8:1:19

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