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Circadian regulation of mTORC1 signaling via Per2 dependent mechanism disrupts folliculogenesis and oocyte maturation in female mice

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Abstract

mTOR (mammalian target of Rapamycin) is an important signaling pathway involved in several crucial ovarian functions including folliculogenesis and oocyte maturation. The circadian rhythm regulates multiple physiological processes and PER2 is one of the core circadian rhythm components. mTOR is regulated by the circadian clock and in turn, the rhythmic mTOR activities strengthen the clock function. Our current study aims to investigate a possible interconnection between the circadian clock and the mTORC1 signaling pathway in folliculogenesis and oocyte maturation. Here we demonstrate that the circadian system regulates mTORC1 signaling via Per2 dependent mechanism in the mouse ovary. To investigate the effect of constant light on ovarian and oocyte morphology, animals were housed 12:12 h L:D group in standard lightening conditions and the 12:12 h L:L group in constant light for one week. Food intake and body weight changes were measured. Ovarian morphology, follicle counting, and oocyte aging were evaluated. Afterward, western blot for mTOR, p-mTOR, p70S6K, p-p70S6K, PER2, and Caspase-3 protein levels was performed. The study demonstrated that circadian rhythm disruption caused an alteration in their food intake and decrease in primordial follicle numbers and an increase in the number of atretic follicles. It caused an increase in oxidative stress and a decrease in ZP3 expression in oocytes. Decreased protein levels of mTOR, p-mTOR, p70S6K, and PER2 were shown. The results showed that the circadian clock regulates mTORC1 through PER2 dependent mechanism and that decreased mTORC1 activity can contribute to premature aging of mouse ovary. In conclusion, these results suggest that the circadian clock may control ovarian aging by regulating mTOR signaling pathway through Per2 expression.

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The authors confirm that the data supporting the findings of this study are available within the article.

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Abbreviations

Bmal1:

Brain and muscle ARNT-like protein 1

Clock:

Circadian locomotor output cycles kaput

Cry1:

Cryptochrome circadian regulator 1

Cry2:

Cryptochrome circadian regulator 2

L:D:

Light-dark cycle

L:L:

Light-light cycle

mTORC1:

The mammalian target of rapamycin complex 1

NTY:

Nitrotyrosine

p-mTOR:

Phosphorylated mammalian target of rapamycin

Per1:

Period Circadian Clock 1

Per2:

Period Circadian Clock 2

Per3:

Period Circadian Clock 3

P70 S6K:

Ribosomal protein S6 Kinase

p-P70 S6K:

Phosphorylated ribosomal protein S6 Kinase

ROS:

Reactive Oxygen Species

TSC1:

Tuberous Sclerosis Complex 1

ZP3:

Zona pellucida sperm binding protein 3

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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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All the authors listed have significantly contributed to the manuscript and there is no disclosure of any conflict of interest.

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Authors and Affiliations

  1. Department of Histology and Embryology, Yeditepe University Faculty of Medicine, 34755, İstanbul, Turkey

    Gizem Bora, Tuğçe Önel, Ecem Yıldırım & Aylin Yaba

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  1. Gizem Bora
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  2. Tuğçe Önel
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  3. Ecem Yıldırım
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  4. Aylin Yaba
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Contributions

A.Y. conceived and coordinated the study, and analyzed experiments. G.B designed experimental groups and performed the analysis of food intake and body weight changes, morphological evaluation of ovaries. G.B. and E.Y. performed follicle counting. G.B., T.O., and E.Y. performed immunofluorescence staining and image analysis. G.B. and T.O. performed a western blot. A.Y. and G.B. wrote the paper.

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Correspondence to Aylin Yaba.

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Bora, G., Önel, T., Yıldırım, E. et al. Circadian regulation of mTORC1 signaling via Per2 dependent mechanism disrupts folliculogenesis and oocyte maturation in female mice. J Mol Histol 54, 217–229 (2023). https://doi.org/10.1007/s10735-023-10126-9

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