Lcn5在山羊睾丸和附睾中的表达特性及定位研究

发布时间：2018-05-05 20:08

本文选题：Lcn5 + 表达特性　；参考：《山西农业大学》2015年硕士论文

【摘要】：Lcn5是Lipocalin家族蛋白成员之一,与雄性动物的生殖密切相关。目前关于Lcn5在生殖器官中的研究主要在大鼠、小鼠中有报道,关于山羊不同发育阶段生殖器官中Lcn5的表达定位及分布规律尚未见报道,因此本试验旨在研究Lcn5在山羊不同发育阶段睾丸和附睾中的表达特性及定位,探讨其对山羊生殖机能的作用,为进一步阐明Lcn5对雄性动物生殖的调控机制提供理论基础。本试验选取7日龄、2月、3月、4月、5月、6月、9月、18月龄晋岚绒山羊公羊各4只,以其睾丸、附睾及18月龄山羊的精子为研究对象,利用QRT-PCR技术、Western-blot技术对山羊睾丸和附睾组织中Lcn5基因mRNA和Lcn5蛋白的相对表达量进行分析。利用免疫组化、免疫荧光技术对Lcn5在山羊睾丸、附睾及精子进行定位分析。主要结果如下：(1) QRT-PCR结果表明,Lcn5在山羊体内广泛表达,生殖器官中表达量较高,其余器官中表达量均极低,且在生殖器官中的表达量为附睾头附睾尾附睾体睾丸。不同发育阶段附睾头、附睾体、附睾尾及睾丸中Lcn5 mRNA的表达量在5月龄前随着月龄的升高逐渐升高,5月龄时最高,随后表达量又逐渐降低。(2) Western-blot结果显示,Lcn5在不同时期的山羊睾丸和附睾中的表达量为：附睾头附睾尾附睾体睾丸,不同月龄的山羊附睾头、附睾体、附睾尾和睾丸中Lcn5蛋白的表达趋势与Lcn5 mRNA表达趋势一致。(3)免疫组化及免疫荧光结果显示：Lcn5在山羊睾丸和附睾中均有表达,在睾丸精原细胞和精子细胞中微量表达,在附睾管假复层柱状上皮细胞、纤毛细胞及管腔液中检测到强阳性信号。精子的免疫荧光结果显示Lcn5主要定位于精子的顶体及尾部中段。不同月龄山羊睾丸和附睾的免疫组化光密度结果分析与mRNA和Western-blot结果相一致,推测Lcn5可能在精子发生、成熟、获能和精卵识别等方面发挥重要的作用。
[Abstract]:Lcn5 is one of the members of the Lipocalin family protein, which is closely related to the reproduction of the male animals. At present, the study of Lcn5 in the reproductive organs is mainly in rats and mice. The expression and distribution of Lcn5 in the reproductive organs of different developmental stages of the goat have not been reported. Therefore, the purpose of this experiment is to study the difference of Lcn5 in the goat. The expression and location of the testicles and epididymis in the developmental stage and its role in the reproductive function of goats are discussed in order to provide a theoretical basis for further clarifying the regulation mechanism of Lcn5 on the reproduction of male animals. The test selected 7 days of age, February, March, April, May, June, September, and 18 month old Jin Lan goat rams each with their testicles, epididymis and 18 month old goats. The relative expression of Lcn5 gene mRNA and Lcn5 protein in goats' testis and epididymis was analyzed by QRT-PCR technology and Western-blot technology. The immunofluorescence technique was used to locate Lcn5 in goat testis, epididymis and spermatozoa. The main results were as follows: (1) QRT-PCR results showed that Lcn5 In the goat, the expression of the reproductive organs is high, the expression of the other organs is very low, and the expression in the reproductive organs is the epididymal testis of the epididymis. The expression of Lcn5 mRNA in the epididymis, epididymis, epididymal tail and testis of different developmental stages increases with the increase of the age of 5 month old, 5 month old (2) Western-blot results showed that the expression of Lcn5 in goat testis and epididymis at different periods was: epididymal testis of epididymis, and the expression trend of Lcn5 protein in epididymis head, epididymis, epididymal tail and testicle in different months of age was consistent with the trend of Lcn5 mRNA expression. (3) immune group. The results of chemical and immunofluorescence showed that Lcn5 was expressed in the testicles and epididymis of goats, expressed in the spermatogonial cells and spermatogonial cells, and the strong positive signals were detected in the epididymal tubular epithelial cells, ciliated cells and the lumen fluid. The immunofluorescence results showed that Lcn5 was mainly located in the acrosome and tail of the sperm. The results of immunohistochemistry of the testicles and epididymis of different months of age are consistent with the results of mRNA and Western-blot. It is presumed that Lcn5 may play an important role in spermatogenesis, maturation, capacitation and sperm identification.

【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827

【参考文献】