Regulation of membrane progestin receptors in the zebrafish ovary by gonadotropin, activin, TGF-beta and BMP-15.
Progestin hormones are vital for inducing oocyte maturation in fish by binding to membrane progestin receptors (mPRs). The aim of this study was to examine the expression and regulation of mPRalpha and mPRbeta in zebrafish follicles. First, defolliculated fully grown oocytes were subjected to immunofluorescent staining using anti-mPRalpha and mPRbeta antibodies, and their expression on the oocyte membrane was confirmed. Second, total protein was collected from zebrafish follicles and Western blotting revealed that the level of mPRalpha and mPRbeta increased with follicle development. We have previously shown that several members of the transforming growth factor-beta (TGF-beta) superfamily, including TGF-beta1, activin-A, and bone morphogenetic protein (BMP)-15, regulate oocyte maturation in zebrafish. Therefore, the third major focus of this study was to test if these growth factors, as well as gonadotropins, regulate the expression of mPRs. Overexpression of BMP-15 significantly reduced, while knockdown of BMP-15 increased, mPRbeta levels. However, mPRalpha expression level remained unchanged with BMP-15 overexpression or knockdown. Treatment of follicles with human chorionic gonadotropin (hCG) resulted in an increased in mPRbeta, but not mPRalpha, expression levels. Activin-A induced the expression of mPRalpha and mPRbeta in a dose- and time-dependent manner. On the other hand, TGF-beta1 treatment suppressed the expression of mPRbeta, but not mPRalpha. Taken together, these findings further support the role of mPRs in oocyte maturation and suggest that gonadotropins, BMP-15, activin-A, and TGF-beta1 exert their regulatory effects on oocyte maturation in part by regulating mPR expression.
Department of Biology, York University, 4700 Keele Street, Toronto, Ontario M3J 1P3, Canada.
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Tan
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Zagrodny
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Bernaudo
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Peng
D028341:Activins / Q000008:administration & dosage / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D055430:Bone Morphogenetic Protein 15 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D002462:Cell Membrane; D006063:Chorionic Gonadotropin / Q000008:administration & dosage / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D009866:Oogenesis; D010957:Plasmids; D011372:Progestins / Q000378:metabolism; D020033:Protein Isoforms; D011980:Receptors, Progesterone / Q000378:metabolism*; D011994:Recombinant Proteins; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000008:administration & dosage / Q000378:metabolism*; D015027:Zebrafish / Q000378:metabolism; D029961:Zebrafish Proteins / Q000378:metabolism*
D055430:Bone Morphogenetic Protein 15; D006063:Chorionic Gonadotropin; D016376:Oligonucleotides, Antisense; C511728:Paqr8 protein, zebrafish; D011372:Progestins; D020033:Protein Isoforms; D011980:Receptors, Progesterone; D011994:Recombinant Proteins; D053773:Transforming Growth Factor beta1; D029961:Zebrafish Proteins; C511727:paqr7b protein, zebrafish; D028341:Activins