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Melatonin ( N -acetyl-5-methoxytryptamine) is found in ovarian follicular fluid, and its concentration is closely related to follicular health status. Nevertheless, the molecular mechanisms underlying melatonin function in follicles are uncertain. In this study, melatonin concentration was measured in porcine follicular fluid at different stages of health. The melatonin concentration decreased as the follicles underwent atresia, suggesting that melatonin may participate in the maintenance of follicular health. The molecular pathway through which melatonin may regulate follicular development was further investigated. The pro-apoptotic protein Bim EL (Bcl-2-interacting mediator of cell death-Extra Long), a key protein controlling granulosa cell apoptosis during follicular atresia, was selected as the target molecule. Bim EL was downregulated when porcine granulosa cells were cultured in medium containing 10 -9 M melatonin and isolated cumulus oocyte complexes (COCs) or follicle stimulating hormone (FSH). Interestingly, ERK-mediated phosphorylation was a prerequisite for the melatonin-induced decline in Bim EL , and melatonin only promoted the ubiquitination of phosphorylated Bim EL , and did not affect the activities of the lysosome or the proteasome. Moreover, the melatonin-induced downregulation of Bim EL was independent of its receptor and its antioxidant properties. In conclusion, melatonin may maintain follicular health by inducing Bim EL ubiquitination to inhibit the apoptosis of granulosa cells.

international journal of molecular sciences

Melatonin Promotes Ubiquitination of Phosphorylated Pro-Apoptotic Protein Bcl-2-Interacting Mediator of Cell Death-Extra Long (BimEL) in Porcine Granulosa Cells