Stimulatory Effect of Progesterone on the Expression of Steroidogenic Acute Regulatory Protein in MA-10 Leydig Cells.

Institute for Hormone and Fertility Research, University of Hamburg, D-22529 Hamburg, Germany.
Biology of Reproduction (Impact Factor: 3.32). 04/2003; 68(3):1054-63.
Source: PubMed


The steroidogenic acute regulatory protein (StAR), by virtue of its ability to facilitate the intramitochondrial transport of cholesterol, plays an important role in regulating steroid hormone biosynthesis in steroidogenic cells. In agreement with published data, both StAR expression and progesterone production in MA-10 mouse Leydig tumor cells could be stimulated with hCG and 8Br-cAMP. Addition of aminoglutethimide, an inhibitor of cholesterol side chain cleavage (P450(scc)) enzyme, not only resulted in a drastic inhibition of progesterone production but also in an attenuation of StAR expression in response to either hCG or 8-Br-cAMP. Therefore, we addressed the question of whether progesterone, the end product of the steroidogenic cascade in these cells, could be in a position to regulate the StAR gene expression. In MA-10 cells, we report here that progesterone in microgram amounts can induce StAR gene expression in a time- and dose-dependent manner. StAR expression in response to a maximally effective concentration of progesterone of 10 microg/ml was highest at 6 h and started decreasing thereafter. The effect of progesterone on StAR protein and StAR mRNA induction was mimicked by its synthetic analog, progestin R5020, but not by other steroids, including dexamethasone, estradiol, testosterone, and dihydrotestosterone. Dexamethasone, in contrast, was able to inhibit StAR expression in MA-10 cells. Surprisingly, RU486, a potent antagonist of progesterone and glucocorticoid action, had a stimulatory effect on StAR mRNA levels. Reverse transcription-polymerase chain reaction analysis demonstrated the absence of the classical form of progesterone receptor in MA-10 cells. Thus, for the first time, a direct stimulatory effect of a steroid on StAR gene expression has been demonstrated. Furthermore, these results provide a new insight, indicating that progesterone mediates the activation of StAR expression exerted presumably through a novel, nonclassical progesterone receptor in mouse Leydig cells.

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    • "Steroid hormones mediate their classical or genomic actions by binding to nuclear receptors and engaging a variety of molecular chaperones permitting ultimate binding of the entire complex to steroid response elements on promoters of target genes [8]. Nonclassical or non-genomic actions of steroids, including rapid changes in cytoplasmic protein function without steroid-receptor complex binding to DNA, are mediated by cell membrane receptors such as GPR30, a G-protein coupled protein mediating rapid estrogen action (estrogen membrane receptor: [9]; androgen membrane receptor: [10]; progesterone membrane receptor: [11]). Whether prenatal steroid hormone programming of postnatal ovarian function involves either or both classical and non-classical action is not known. "
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    ABSTRACT: In female mammals, including humans, deviations from normal androgenic or estrogenic exposure during fetal development are detrimental to subsequent adult ovarian function. Androgen deficiency, without accompanying estrogen deficit, has little apparent impact on ovarian development. Fetal estrogen deficiency, on the other hand, results in impaired oocyte and follicle development, immature and abnormal adult ovaries, and excessive ovarian stimulation from endogenous gonadotropins ultimately generating hemorrhagic follicles. Complete estrogen deficiency lasting into adulthood results in partial ovarian masculinization. Fetal androgen excess, on the other hand, mediated either by direct androgen action or following androgen aromatization to estrogen, reprograms ovarian development and reproductive neuroendocrinology to mimic that found in women with polycystic ovary syndrome: enlarged, polyfollicular, hyperandrogenic, anovulatory ovaries with accompanying LH hypersecretion. Oocyte developmental competence is also compromised. Insulin is implicated in the mechanism of both anovulation and deficient oocyte development. Fetal estrogen excess induces somewhat similar disruption of adult ovarian function to fetal androgen excess. Understanding the quality of the fetal female sex steroid hormone environment is thus becoming increasingly important in improving our knowledge of mechanisms underlying a variety of female reproductive pathologies.
    Reproductive Biology and Endocrinology 02/2006; 4(1):17. DOI:10.1186/1477-7827-4-17 · 2.23 Impact Factor
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    • "The mechanism regulating StAR transcription in the absence of a canonical CRE has been demonstrated to be mediated by multiple DNA elements that are found to be located in a transcription factor-binding site-rich region in the StAR promoter (reviewed in Ref. 24). StAR expression has also been influenced by various compounds, including peptide and non-peptide hormones, growth factors, gonadotropin-releasing hormone, prostaglandins and steroids, acting through endocrine, autocrine and paracrine mechanisms (36, 43, 50, 54-56). It has also been demonstrated that phosphorylation of the StAR protein is critical in producing full cholesterol transferring activity (57-58). "
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    ABSTRACT: The steroidogenic acute regulatory (StAR) protein, a novel mitochondrial protein, is involved in the regulation of steroid hormone biosynthesis through its mediation of the intramitochondrial transport of the steroid substrate, cholesterol, to the cytochrome P450 cholesterol side chain cleavage (P450scc) enzyme. The expression of StAR protein is regulated by cAMP-dependent signaling in steroidogenic cells. During the course of our studies in mouse Leydig cells, we employ several methods for studying the regulation of StAR protein expression by human chorionic gonadotropin (hCG). A sensitive quantitative reverse transcription and polymerase chain reaction (RT-PCR) was utilized for determining StAR mRNA expression. Stimulation of mLTC-1 mouse Leydig tumor cells with hCG resulted in the coordinate regulation of StAR mRNA expression and progesterone accumulation in a time-response manner. The validity and accuracy of quantitative RT-PCR results in mLTC-1 cells were verified by a competitive PCR approach and were further confirmed in primary cultures of isolated mouse Leydig cells. Immunoblotting studies demonstrated an increase in the levels of the StAR protein in a concentration dependent manner following hCG stimulation in mLTC-1 cells. Northern hybridization analysis revealed three StAR transcripts, all of which were of sufficient size to encode functional StAR protein, and which were coordinately expressed in response to hCG. Collectively, the experimental approaches utilized in the present investigation allow for the demonstration and characterization of hCG mediated regulation of StAR mRNA and StAR protein expression in mouse Leydig cells.
    Biological Procedures Online 02/2004; 6:83-93. DOI:10.1251/bpo76 · 1.79 Impact Factor
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    • "The mechanisms by which these factors act to control StAR levels, by and large, have not been elucidated but may encompass transcriptional as well as post-transcriptional mechanisms. Interestingly, progesterone was recently shown to have a stimulatory effect on StAR gene expression in a mouse Leydig cell line (MA-10) through a yet to be determined mechanism [78]. This mechanism does not require classical progesterone receptors as MA-10 cells are devoid of this receptor [78]. "
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    Reproductive Biology and Endocrinology 12/2003; 1(1):90. DOI:10.1186/1477-7827-1-90 · 2.23 Impact Factor
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