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Equine chorionic gonadotropin influence on sheep oocyte in vitro maturation, apoptosis, and follicle-stimulating hormone receptor and luteinizing hormone receptor expression

机译:马绒毛膜促性腺激素对绵羊卵母细胞体外成熟,凋亡,促卵泡激素受体和促黄体生成激素受体表达的影响

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We assessed the effects of equine chorionic gonadotropin (eCG) on oocyte in vitro maturation (IVM), apoptosis, and follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), and gonadotropin-releasing hormone receptor (GnRHR) expression and mRNA levels. Cumulus-oocyte complexes (COCs) were recovered from sheep ovaries and pooled in groups, before being cultured in IVM media containing varying eCG concentrations. Maturation and apoptosis rates were then calculated. Expression of FSHR, LHR, and GnRHR mRNA in oocytes was measured using quantitative reverse transcription polymerase chain reaction. Protein levels were ascertained by western blotting. Matured oocytes displayed and released an intact first polar body. Sheep oocyte maturation rates gradually increased as eCG concentration was raised from 0 to 20 μg/mL. Apoptosis rates of eCG-treated oocytes were lower than those of the control group, and were lowest using 20 μg/mL eCG. FSHR, LHR, and GnRHR mRNA expression increased (P 0.01, P 0.05, and P 0.05, respectively, compared to 0 μg/mL eCG) with eCG concentration, being highest following exposure to 20 μg/mL. FSHR and GnRHR protein levels were significantly higher in oocytes administered 20 μg/mL eCG compared with those matured in the absence of eCG. eCG dose positively correlated with FSHR, LHR, and GnRHR mRNA and protein expression. In conclusion, eCG enhances maturation and decreases apoptosis of oocytes undergoing IVM, and heightens FSHR, LHR, and GnRHR expression. Such increased expression may facilitate oocyte IVM. These findings contribute to our understanding of the mechanisms of underlying hormonal control of sheep oocyte IVM, advancing ovine reproductive methods.
机译:我们评估了马绒毛膜促性腺激素(eCG)对卵母细胞体外成熟(IVM),细胞凋亡和促卵泡激素受体(FSHR),促黄体激素受体(LHR)和促性腺激素释放激素受体(GnRHR)的影响,以及mRNA水平。从绵羊卵巢中回收卵母细胞复合物(COCs)并成组合并,然后在含有不同eCG浓度的IVM培养基中培养。然后计算成熟度和凋亡率。使用定量逆转录聚合酶链反应测量卵母细胞中FSHR,LHR和GnRHR mRNA的表达。通过蛋白质印迹确定蛋白质水平。成熟的卵母细胞显示并释放完整的第一极体。随着eCG浓度从0增加到20μg/ mL,绵羊卵母细胞成熟率逐渐增加。用eCG处理的卵母细胞的凋亡率低于对照组,并且使用20μg/ mL eCG时最低。 FSHR,LHR和GnRHR mRNA表达随eCG浓度的增加而增加(分别为P <0.01,P <0.05和P <0.05,与0μg/ mL eCG相比),在暴露于20μg/ mL后最高。与没有eCG时成熟的卵母细胞相比,给予20μg/ mL eCG的卵母细胞中的FSHR和GnRHR蛋白水平明显更高。 eCG剂量与FSHR,LHR和GnRHR mRNA和蛋白质表达呈正相关。总之,eCG增强了经历IVM的卵母细胞的成熟并减少其凋亡,并提高了FSHR,LHR和GnRHR的表达。这种增加的表达可以促进卵母细胞IVM。这些发现有助于我们对绵羊卵母细胞IVM的基本激素控制机制的理解,从而促进绵羊的生殖方法。

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