蛋鸡不同发育阶段卵巢与卵泡中ERα和ERβ的表达和分布研究

发布时间：2018-05-07 10:13

本文选题：鸡 + 卵巢　；参考：《山东农业大学》2015年硕士论文

【摘要】：禽类卵巢卵泡的发育是一个复杂且精细的过程,不断经历着卵泡的生长、优势卵泡的选择和卵泡分化等一系列重要环节。另外,卵巢卵泡的发育又受到内分泌、自分泌、旁分泌等多种方式的调节。在畜牧业生产中,研究禽类卵巢以及卵泡发育调控的机制对于有效地开发利用卵巢内的卵泡资源,提高家禽的繁殖性能,改善蛋禽的产蛋性能,提高生产效率具有重要意义。在卵泡生长过程中,雌激素是一种关键的卵泡内调节分子,是由颗粒细胞和卵泡内膜细胞合成的类固醇类激素的一种,具有加快卵泡内颗粒细胞的增殖、增强FSH与黄体生成素LH对卵巢细胞的增殖与分化产生效应的作用,另外雌激素也可以调节卵泡的等级发育。总之,雌激素在动物繁殖发育的过程中起着非常重要的作用,而大多数的雌激素是通过两种不同的核内雌激素受体ERα与ERβ来发挥作用的。ERα与ERβ是主要的依赖配体活化转录因子的蛋白超家族成员,通过参与转录过程调控靶细胞的增殖和分化。目前,ERα与ERβ基因mRNA在鸡不同等级卵泡中的表达情况已有部分研究,但是在不同时期鸡卵巢与卵泡中ERα与ERβ基因mRNA和蛋白的表达及定位情况尚未见报导。为了进一步了解ERα与ERβ在鸡卵泡发育过程中的潜在作用,运用q RT-PCR和Western blotting技术分别检测了45、60、90、110、120与160天鸡卵巢与卵泡中ERα与ERβmRNA和蛋白的表达。结果显示ERα基因mRNA呈现V样表达模式,从45天到90天逐渐降低,并且在90天时最低,从90天到160天逐渐增加。而且45、160天时ERαmRNA显著高于90天,蛋白表达与mRNA表达趋势基本一致。免疫组化分析揭示了45天鸡卵巢中ERα主要富集于卵巢初级卵泡的颗粒细胞层和卵母细胞,在细胞膜和细胞间质有少量表达,而且发现在卵巢髓质层的血管中有一定量表达证明ERα在血管形成中起作用。在各级卵泡中ERαmRNA和蛋白在SY卵泡中表达最高,而在SY、F5、F3、F1各级卵泡中呈现逐级递减趋势,免疫组化结果显示ERα蛋白主要分布在颗粒细胞层,而且SW、SY卵泡中表达相对较高,在F5、F3、F1中表达相对较低。这结果暗示,在发育过程中雌激素很可能是通过自分泌/旁分泌形式与ERα结合进行卵巢卵泡功能的调控。而ERβmRNA呈现L样表达模式,在45天表达量最高,60天到90天急速下降,90天之后表达较低并趋于稳定,蛋白表达呈现逐渐降低趋势。研究显示ERβmRNA和蛋白在SW到F1的各级卵泡中逐渐降低。结果表明不同时期鸡卵巢及各级卵泡中ERα与ERβ基因表达存在明显的不同。这暗示两类受体对蛋鸡卵巢与卵泡的发育具有不同的作用,具体作用机制有待进一步研究。
[Abstract]:The development of poultry ovarian follicles is a complex and fine process, which constantly experiences a series of important steps, such as follicular growth, selection of dominant follicles and follicular differentiation. In addition, ovarian follicle development by endocrine, autocrine, paracrine and other ways of regulation. In animal husbandry, it is of great significance to study the mechanism of ovaries and follicle development regulation in poultry for the effective development and utilization of follicular resources in ovaries, the improvement of poultry reproductive performance, the improvement of egg laying performance, and the improvement of production efficiency. During follicular growth, estrogen is a key regulatory molecule in the follicle, a steroid hormone synthesized by granulosa cells and endometrial cells of the follicle, which accelerates the proliferation of granulosa cells in the follicle. The effects of FSH and luteinizing hormone LH on the proliferation and differentiation of ovarian cells were enhanced. In addition, estrogen could also regulate the grade development of follicles. In short, estrogen plays a very important role in animal reproduction and development. The majority of estrogen is mediated by two different nuclear estrogen receptors, ER 伪 and ER 尾. ER 伪 and ER 尾 are major ligand-activated transcription factor dependent protein superfamily members. The proliferation and differentiation of target cells are regulated by participating in the transcription process. At present, the expression of ER 伪 and ER 尾 gene mRNA in different grade follicles has been partly studied, but the expression and localization of ER 伪 and ER 尾 gene mRNA and protein in different stages of chicken ovary and follicle have not been reported. In order to further understand the potential role of ER 伪 and ER 尾 in the development of chicken follicles, the expression of ER 伪 and ER 尾 mRNA and protein in the ovary and follicles were detected by Q RT-PCR and Western blotting techniques respectively. The results showed that the expression pattern of ER 伪 gene mRNA was V-like, which decreased gradually from 45 days to 90 days, and increased gradually from 90 days to 160 days after 90 days. The expression of ER 伪 mRNA was significantly higher than that of 90 days at day 45160, and the expression of ER 伪 mRNA was consistent with the trend of mRNA expression. Immunohistochemical analysis revealed that ER 伪 was mainly concentrated in granulosa cell layer and oocyte of primary follicle of ovary at 45 days, and expressed a little in cell membrane and stroma. It was also found that ER 伪 plays an important role in angiogenesis in the medullary layer of ovary. The expression of ER 伪 mRNA and protein was the highest in sy follicles, but decreased gradually in SYF5F3F3F 1 follicles. The immunohistochemical results showed that ER 伪 protein was mainly distributed in granulosa layer, and the expression of ER 伪 protein was relatively high in SWSY follicles. The expression was relatively low in F _ 5, F _ 3, F _ 1. These results suggest that estrogen may regulate ovarian follicle function through the combination of ER 伪 and autocrine / paracrine during development. On the other hand, the expression of ER 尾 mRNA showed L-like expression pattern. The expression of ER 尾 mRNA decreased rapidly from 60 days to 90 days after 45 days. The expression of ER 尾 mRNA tended to be stable, and the expression of protein decreased gradually. The results showed that ER 尾 mRNA and protein decreased gradually in all follicles from SW to F 1. The results showed that ER 伪 and ER 尾 gene expression were significantly different in different stages of chicken ovary and follicles. This suggests that the two receptors have different effects on the development of ovaries and follicles in laying hens, and the specific mechanisms need to be further studied.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831

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