决定卵母细胞能力的重要事件发生在卵泡生长和卵母细胞成熟期间。抗苗勒管激素(AMH),生育率的积极预测,已被证明受暴露于内分泌干扰化合物(EDC)的影响,如双酚A(BPA)和S(BPS)。然而,双酚和SMAD蛋白之间的相互作用,AMH途径的介质,尚未阐明。在用BPA处理的牛颗粒细胞中研究AMH受体(AMHRII)和下游SMADs的表达,BPS,然后与AMH竞争。在这里,我们显示颗粒细胞中24小时BPA暴露显着增加SMAD1,SMAD4和SMAD5mRNA表达。在24小时处理后,在AMHRII或SMADs蛋白表达中没有观察到显著变化。用BPA(单独或与AMH一起)治疗12小时后,观察到SMAD1和SMAD4mRNA表达显着增加,而在蛋白质水平上检测到SMAD1和磷酸化SMAD1的显着降低。建立双酚与AMH信号通路之间的功能性联系,反义寡核苷酸(ASO)用于在有或没有双酚暴露的情况下抑制AMHRII表达。最初,优化和验证转染条件,实现70%敲低。我们的发现表明,BPS发挥其作用独立于AMH受体,而BPA可能直接通过AMH信号通路起作用,提供了一种潜在的机制,双酚可能通过该机制发挥其作用来破坏卵泡发育并降低卵母细胞能力。
Significant events that determine oocyte competence occur during follicular growth and oocyte maturation. The anti-Mullerian hormone, a positive predictor of fertility, has been shown to be affected by exposure to endocrine disrupting compounds, such as bisphenol A and S. However, the interaction between bisphenols and SMAD proteins, mediators of the anti-Mullerian hormone pathway, has not yet been elucidated. AMH receptor (
AMHRII) and downstream SMAD expression was investigated in bovine granulosa cells treated with bisphenol A, bisphenol S, and then competitively with the anti-Mullerian hormone. Here, we show that 24-h bisphenol A exposure in granulosa cells significantly increased SMAD1, SMAD4, and SMAD5 mRNA expression. No significant changes were observed in
AMHRII or SMADs protein expression after 24-h treatment. Following 12-h treatments with bisphenol A (alone or with the anti-Mullerian hormone), a significant increase in SMAD1 and SMAD4 mRNA expression was observed, while a significant decrease in SMAD1 and phosphorylated SMAD1 was detected at the protein level. To establish a functional link between bisphenols and the anti-Mullerian hormone signaling pathway, antisense oligonucleotides were utilized to suppress
AMHRII expression with or without bisphenol exposure. Initially, transfection conditions were optimized and validated with a 70% knockdown achieved. Our findings show that bisphenol S exerts its effects independently of the anti-Mullerian hormone receptor, while bisphenol A may act directly through the anti-Mullerian hormone signaling pathway providing a potential mechanism by which bisphenols may exert their actions to disrupt follicular development and decrease oocyte competence.