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

Sep 12, 2016Journal of ovarian research

Changes in daily rhythm and hormone-making genes in the human ovary after testosterone exposure

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Abstract

Immunostaining revealed the presence of CLOCK and PER2 proteins in granulosa cells of dominant antral follicles in human ovaries.

Circadian genes CLOCK and PER2 are specifically expressed in dominant antral follicles but not in primordial, primary, or preantral follicles.
Testosterone treatment led to an oscillating expression pattern of PER2, with peaks at 24 and 44 hours.
CLOCK expression significantly increased, peaking at 24 hours post-testosterone treatment, while BMAL1 expression remained unchanged.
Among steroidogenesis-related genes, only STAR exhibited oscillating expression after testosterone stimulation, with peaks at 24 and 40 hours.
These findings suggest a potential link between circadian rhythms and steroid hormone production in the human ovary.

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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.

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

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