猪ESR、FSHβ和Leptin基因多态性及其与卵泡囊肿的相关性研究

发布时间：2018-06-15 01:25

本文选题：卵泡囊肿 + 雌激素受体　；参考：《吉林大学》2017年硕士论文

【摘要】：卵泡囊肿是母猪常见的繁殖障碍性疾病之一,会导致母猪不孕。它会引起母畜持续、频繁、短暂和无规律的发情,该病对猪饲养产业的发展造成了巨大的阻碍。现阶段,随着分子生物学的研究愈加深入,寻找猪卵泡囊肿标记基因的研究也越来越受重视。ESR、FSHβ和Leptin基因均可以通过调控下丘脑-垂体-卵巢轴的每个环节从而对母猪卵巢生长发育起到重要的作用。ESR(雌激素受体)是类固醇受体家族的成员之一,它能够调控雌性哺乳动物的生殖活动,影响雌性动物性腺组织基因的表达和调控,进一步影响动物个体的第二性征,繁殖周期,生殖力,妊娠维持等。有研究结果表明猪ESR基因PvuⅡ多态性酶切位点对猪的产仔数和繁殖性状有显著影响。然而目前尚无关于分析该位点与卵泡囊肿关系的报道。FSH(卵泡刺激素)是一种糖蛋白类促性腺激素,对促进颗粒细胞成熟有重要的作用。猪的FSH基因结构包含一长度为292bp的逆转座子,由于逆转座子的插入而导致了该基因位点存在多态性。经过对该位点的分析发现,FSHβ基因是影响猪繁殖性状的主效基因。但是目前尚无研究把FSHβ基因作为侯选基因来分析其与卵泡囊肿的关系。Leptin(瘦素)是由脂肪细胞分泌的一类蛋白质类激素,其基因表达产物主要参与动物生长发育和繁殖功能等方面的调节。有研究表明,猪leptin基因第3外显子T3469C位点变异对猪的繁殖性能具有一定影响。然而,尚无研究将Leptin基因作为侯选基因来分析其与卵泡囊肿的关系。本实验以雌激素受体(ESR)、促卵泡素β亚基(FSHβ)和瘦素(Leptin)作为研究对象。收集健康卵巢和患病卵巢后,通过提取卵巢基因组DNA,然后采用直接电泳法和PCR-RFLP法对雌激素受体(ESR)、促卵泡素β亚基(FSHβ)和瘦素(Leptin)进行基因多态位点检测,并进一步分析该位点与卵泡囊肿的关系,寻找与卵泡囊肿有关的分子标记。主要内容如下:(1)基因多态性检测结果显示:3个基因共检测到4个多态位点,分别是位于ESR基因内含子AG突变;位于FSHβ基因内含子5’调控区TC突变;位于FSHβ基因内含子CT突变;位于leptin基因外显子TC突变。(2)多态信息含量、杂合度和有效等位基因分析结果显示:ESR PvuⅡ多态酶切位点在两群体中均呈现中度多态(0.25PIC0.5),该多态位点基因型分布在囊肿群体中背离Hardy-Weinberg平衡状态(P0.05),在健康猪群体中符合Hardy-Weinberg平衡(P0.05);FSHβ5’调控区多态位点在卵泡囊肿猪群体中呈现低度多态(PIC0.25),健康猪群体中均呈现中度多态(0.25PIC0.5)。该多态位点基因型分布两群体中均背离Hardy-Weinberg平衡状态(P0.05);FSHβ-HaeⅢ多态酶切位点在两群体中均呈现低度多态(PIC0.25),多态水平较低。该多态位点基因型分布均符合Hardy-Weinberg平衡状态(P0.05);Leptin基因Hinf I酶切位点在两群体中均处于低度多态(PIC0.25),多态水平较低。该多态位点基因型分布在卵泡囊肿群体符合Hardy-Weinberg平衡状态(P0.05),在健康猪群体背离Hardy-Weinberg平衡状态(P0.05)。(3)利用分析软件对ESR、FSHβ和Leptin的基因型分布进行分析,结果表明ESR的AA和AB基因型在健康群体和卵泡囊肿群体分布差异显著(p0.05),与卵泡囊肿的形成有关;FSHβ基因和leptin基因的基因型分布在本检测群体中均未见显著差异(p0.05),与卵泡囊肿的形成无关。ESR基因可作为卵泡囊肿筛选的标志基因,为研发提高母猪繁殖力的新技术奠定基础。
[Abstract]:Follicular cyst is one of the common reproductive disorders of sow, which can cause infertile sows. It can cause maternal animals to continue, frequent, transient and irregular. The disease has caused great obstacles to the development of pig breeding industry. At present, the more research of molecular biology is being made, the more research on the marker gene of porcine follicle cyst is found. The more emphasis is placed on.ESR, the FSH beta and Leptin genes can play an important role in the growth and development of the sow ovary by regulating each link of the hypothalamus pituitary ovary axis..ESR (estrogen receptor) is one of the members of the steroid receptor family. It can regulate the reproductive activities of female mammals and affect the sex gland tissue of female animals. The expression and regulation of these factors further affect the second sex, reproductive cycle, fertility, and pregnancy maintenance of the individual animal. The results show that the ESR gene Pvu II polymorphism of the pig gene has a significant influence on the litter size and reproductive traits of pigs. However, there is no report on the relationship between this site and follicle cyst,.FSH (follicle stimulation) at present. It is a glycoprotein type gonadotropin which plays an important role in promoting the maturation of granular cells. The FSH gene structure of the pig contains a reverse transposon of a length of 292bp, which leads to the polymorphism of the gene site due to the insertion of the transposon. After the analysis of this site, the FSH beta gene is the main effect on the reproductive traits of pigs. But there is no study of the FSH beta gene as a candidate gene to analyze its relationship with follicle cysts..Leptin (leptin) is a class of protein hormones secreted by adipocytes. Its gene expression products are mainly involved in the regulation of animal growth and reproductive function. Some studies have shown that the third exon T3469C of Porcine leptin gene is T3469C Loci variation has a certain effect on the reproductive performance of pigs. However, no study has been made to analyze the relationship between the Leptin gene as a candidate gene and the follicle cyst. This experiment was conducted by estrogen receptor (ESR), follicle stimulating hormone beta subunit (FSH beta) and leptin (Leptin). After collecting healthy and sick ovaries, the ovarian genes were extracted by extracting the ovarian genes. Group DNA, then use direct electrophoresis and PCR-RFLP method to detect the polymorphic loci of estrogen receptor (ESR), follicle stimulating hormone beta subunit (FSH beta) and leptin (Leptin), and further analyze the relationship between the loci and follicle cysts, and find the molecular markers related to follicle cysts. The main contents are as follows: (1) the results of gene polymorphism detection show: 3 A total of 4 polymorphic loci were detected in the ESR gene intron AG mutation, located in the TC mutation in the intron of the FSH beta gene and the CT mutation in the intron of the FSH beta gene, and the TC mutation in the exon of the leptin gene. (2) the polymorphism information content, heterozygosity and effective allele analysis showed that the ESR Pvu II polymorphic loci Moderate polymorphism (0.25PIC0.5) was present in the two population, which was distributed in the equilibrium state of Hardy-Weinberg in the cyst population (P0.05), and in the healthy pig population, which was in line with Hardy-Weinberg balance (P0.05), and the polymorphic loci of the FSH beta 5 'regulatory region showed low polymorphism (PIC0.25) in the follicular cysts, and all of the healthy pigs were all A moderate polymorphism (0.25PIC0.5). The polymorphism loci of the polymorphic loci were all deviated from the equilibrium state of Hardy-Weinberg (P0.05), and the FSH beta -Hae III polymorphic loci were low polymorphic in the two population (PIC0.25), and the polymorphism level was lower than that of Hardy-Weinberg equilibrium state (P0.05) and Leptin gene Hin. The f I loci were low polymorphism (PIC0.25) in the two population, and the polymorphic loci were distributed in the follicle cyst group with Hardy-Weinberg equilibrium state (P0.05) and Hardy-Weinberg balance in healthy pig population (P0.05). (3) the distribution of the genotype distribution of ESR, FSH beta and Leptin was divided by the use analysis software. The results showed that the distribution of AA and AB genotypes of ESR in healthy population and follicle cyst was significantly different (P0.05), which was related to the formation of follicle cysts. The genotype distribution of FSH beta gene and leptin gene was not significantly different in this test group (P0.05). It was not related to the formation of follicle swollen, which could be used as the marker of follicle cyst screening. Chi gene is the foundation for developing new technology to improve sow's fecundity.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S828

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