Journal
MOLECULAR REPRODUCTION AND DEVELOPMENT
Volume 89, Issue 10, Pages 485-497Publisher
WILEY
DOI: 10.1002/mrd.23639
Keywords
hormone receptor; immunohistochemistry; in vitro culture; ovarian cortex; preantral follicle
Funding
- Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico
- Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior
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This study investigated the expression of melatonin receptor type 1 (MT1) protein in sheep ovaries and evaluated the effects of melatonin on the survival and development of primordial follicles after in vitro culture. The results showed that 100 pg/ml melatonin could increase follicular survival and activation. Inhibition of the PI3K pathway resulted in increased apoptosis and decreased activation of primordial follicles. These findings suggest that 100 pg/ml melatonin can promote the survival and activation of primordial follicles through the PI3K signaling pathway in sheep ovarian tissue culture.
This study characterized the expression of melatonin receptor type 1 (MT1) protein in sheep ovaries, evaluated melatonin effects on primordial follicle survival and development after in vitro culture of ovarian tissue and verified the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B/forkhead box O3a (PI3K/Akt/FOXO3a) pathway in the melatonin actions. Ovine ovarian fragments were cultured in alpha-modified minimum essential medium alone (alpha-MEM+) or supplemented with 100, 500, or 1000 pg/ml melatonin for 7 days. PI3K inhibition was performed through pretreatment of ovarian fragments with LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3, Akt, phosphorylated-Akt, and phosphorylated-FOXO3a (p-FOXO3a). The immunohistochemical localization of the MT1 receptor protein was documented in sheep preantral and antral follicles. After in vitro culture, 100 pg/ml melatonin showed higher follicular survival and activation than alpha-MEM+ and other melatonin concentrations. After PI3K inhibition, there was an increase in cleaved caspase-3-positive follicles, and a decrease in the primordial follicle activation, Akt phosphorylation, and nuclear exclusion of p-FOXO3a. In conclusion, MT1 receptor protein is present in the sheep ovary. Furthermore, 100 pg/ml melatonin maintains survival and stimulates activation of primordial follicles through the PI3K/Akt/FOXO3a signaling pathway after in vitro culture of sheep ovarian tissue.
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