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Membrane receptor-independent inhibitory effect of melatonin on androgen production in porcine theca cells

Wang, Heng, Pu, Yong, Luo, Lei, Li, Yunsheng, Zhang, Yunhai, Cao, Zubing
Theriogenology 2018 v.118 pp. 63-71
androstenedione, antagonists, biosynthesis, female fertility, flow cytometry, fluorescent antibody technique, genes, melatonin, ovarian follicles, progesterone, quantitative polymerase chain reaction, radioimmunoassays, receptors, reverse transcriptase polymerase chain reaction, secretion, steroidogenesis, swine, testosterone, viability
Excessive secretion of androgens including androstenedione and testosterone in theca cells frequently causes female infertility in mammals. Melatonin is a potent inhibitor of androgen production in gonadal cells of several species in a membrane receptor-dependent manner. However, the function of melatonin in steroidogenesis of porcine theca cells remains unclear. Here we report that melatonin inhibits androgen biosynthesis independently of its membrane receptors in pigs. Using flow cytometry, immunofluorescence and RT-PCR we showed that the vast majority of cells isolated from the theca layer of antral follicles are indeed theca cells. Furthermore, we demonstrated that of the two of melatonin membrane receptors encoded in the porcine genome, theca cells exclusively express melatonin receptor 1B. Cell counting analysis indicated that different concentrations of melatonin did not alter the normal viability and proliferation of theca cells. Additionally, hormone radioimmunoassay and qPCR respectively showed that a high concentration of melatonin significantly repressed both androgen production and expression of steroidogenic genes involving StAR, CYP11A1, HSD3β and SET (P < 0.05), but did not impair progesterone production. Interestingly, these effects were not reversed by N-acetyl-2-benzyltryptamin, a melatonin membrane receptor antagonist. Overall, these results demonstrate that melatonin inhibits androgen production in porcine theca cells independently of its membrane receptor.