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Background  Previous studies have shown that circadian genes might be involved in the development of polycystic ovarian syndrome (PCOS). Hyperandrogenism is a hallmark feature of PCOS. However, the effect of hyperandrogenism on circadian gene expression in human granulosa cells is unknown, and the general expression pattern of circadian genes in the human ovary is unclear.    Methods  Expression of the circadian proteins CLOCK and PER2 in human ovaries was observed by immunohistochemistry. The mRNA expression patterns of the circadian genes  CLOCK ,  PER2 , and  BMAL1 , and the steroidogenesis-related genes  STAR ,  CYP11A1 ,  HSD3B2 , and  CYP19A1  in cultured human luteinized granulosa cells were analyzed over the course of 48 h after testosterone treatment by quantitative polymerase chain reaction.    Results  Immunostaining of CLOCK and PER2 protein was detected in the granulosa cells of dominant antral follicles but was absent in the primordial, primary, or preantral follicles of human ovaries. After testosterone stimulation, expression of  PER2  showed an oscillating pattern, with two peaks occurring at the 24th and 44th hours; expression of  CLOCK  increased significantly to the peak at the 24th hour, whereas expression of  BMAL1  did not change significantly over time in human luteinized granulosa cells. Among the four steroidogenesis-related genes evaluated, only  STAR  displayed an oscillating expression pattern with two peaks occurring at the 24th and 40th hours after testosterone stimulation.    Conclusions  Circadian genes are expressed in the dominant antral follicles of the human ovary. Oscillating expression of the circadian gene  PER2  can be induced by testosterone in human granulosa cells in vitro. Expression of  STAR  also displayed an oscillating pattern after testosterone stimulation. Our results indicate a potential relationship between the circadian clock and steroidogenesis in the human ovary, and demonstrate the effect of testosterone on circadian gene expression in granulosa cells.

Expression pattern of circadian genes and steroidogenesis-related genes after testosterone stimulation in the human ovary