绵羊发情周期内类固醇合成相关基因的表达变化

发布时间：2018-03-15 18:31

本文选题：黄体　切入点：类固醇　出处：《内蒙古农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：黄体是哺乳动物卵巢上重要的功能单位,与动物正常生殖的维持密切相关,其功能性变化与动物的发情周期密切相关,类固醇合成相关基因StAR、P450(CYP11A)、 3β-HSD在卵巢的周期性变化过程中具有积极作用。因此,了解绵羊卵巢周期性变化过程中黄体形成类固醇合成相关基因与潜在能量代谢通路AMPK、ERK1/2的关系对进一步认识卵巢功能具有价值。为此,本研究以绵羊发情周期卵巢为材料,应用RT-qPCR、Western Blot和免疫荧光(三标)技术,分析发情后第0、3、6、9、12d的绵羊卵巢中类固醇激素合成相关基因的表达变化,以期了解它们在绵羊卵巢发情周期不同时期的表达规律及能量代谢与激素合成相关基因之间潜在的联系。结果表明,发情后第0、3、6、9、12d,绵羊卵巢黄体组织类固醇激素StAR、3β-HSD mRNA的表达量呈逐步升高然后降低的趋势,在黄体上的表达量第9d达到最高(p0.05); 3β-HSD蛋白水平在黄体上的变化较卵巢显著(p0.01),呈上升趋势。而CYP11AmRNA水平随发情周期的延续在第9到第12d仍呈逐渐升高的趋势。同时,在已形成黄体的卵巢和未形成黄体的卵巢上GC、卵母细胞、各级卵母细胞及黄体上均有3β-HSD、StAR蛋白的表达,特别指出的是(CYP17A1蛋白在卵母细胞呈阴性。且类固醇激素合成相关基因StAR、P450(CYP11A)、3p-HSD mRNA相对表达量与血液孕酮水平一致。AMPK和ERK1/2的磷酸化在发情周期的卵巢上呈逐渐升高然后下降的显著变化。类固醇合成相关基因StAR、P450(CYP11A)、3β-HSD参与绵羊发情周期变化的进程,且与AMPK和ERK1/2通路积极相关的实验结果提示,类固醇激素合成相关基因与卵泡发育和黄体功能的变化密切相关,该结果对进一步认识绵羊繁殖过程的调节机制奠定了基础。
[Abstract]:Luteal body is an important functional unit in mammalian ovary, which is closely related to the maintenance of normal reproduction, and its functional changes are closely related to the oestrous cycle of animals. Steroid biosynthesis related gene Starn P450 CYP11AU, 3 尾 -HSD plays an active role in the periodic changes of ovary. It is valuable to understand the relationship between luteal steroid biosynthesis related genes and potential energy metabolism pathway AMPK ERK1 / 2 during the periodic changes of ovaries in sheep. Therefore, the ovaries of sheep estrous cycle were used as materials. The expression changes of steroid hormone synthesis-related genes in ovaries of sheep were analyzed by RT-qPCRG Western Blot and immunofluorescence (triple labeling) technique on the 12th day after oestrus. In order to understand their expression at different stages of oestrous cycle in sheep and the potential relationship between energy metabolism and hormone biosynthesis genes, the results showed that, On the 12th day after oestrus, the expression of steroid steroid hormone Starn3 尾 -HSD mRNA in ovarian luteal tissue of sheep increased gradually and then decreased. The expression level of 3 尾 -HSD on the corpus luteum reached the highest level on the 9th day, and the level of 3 尾 -HSD protein in the corpus luteum was significantly higher than that in the ovary, and the level of CYP11AmRNA increased gradually with the continuation of the estrus cycle from the 9th to the 12th day. At the same time, the expression of 尾 -HSD protein in the corpus luteum was significantly higher than that in the ovary. The expression of 3 尾 -HSD-Star protein was observed in the ovaries of luteal formation and ovaries without luteal formation, and in oocytes, oocytes and corpus luteum. It was pointed out that CYP17A1 protein was negative in oocytes, and the relative expression of steroid hormone synthesis-associated gene StARP450, CYP11Af3p-HSD mRNA was consistent with the level of blood progesterone. Phosphorylation of AMPK and ERK1/2 increased gradually in oocytes during estrous cycle and then decreased. The steroid biosynthesis related gene Starn P450, CYP11Af3 尾 -HSD, was involved in the process of estrous cycle change in sheep. The results of positive correlation with AMPK and ERK1/2 pathway suggested that steroid hormone biosynthesis related genes were closely related to follicular development and luteal function, which laid a foundation for further understanding the regulation mechanism of sheep reproductive process.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826

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