本文選題：Lcn5 + 表達特性�。� 參考：《山西農業(yè)大學》2015年碩士論文

【摘要】：Lcn5是Lipocalin家族蛋白成員之一,與雄性動物的生殖密切相關。目前關于Lcn5在生殖器官中的研究主要在大鼠、小鼠中有報道,關于山羊不同發(fā)育階段生殖器官中Lcn5的表達定位及分布規(guī)律尚未見報道,因此本試驗旨在研究Lcn5在山羊不同發(fā)育階段睪丸和附睪中的表達特性及定位,探討其對山羊生殖機能的作用,為進一步闡明Lcn5對雄性動物生殖的調控機制提供理論基礎。本試驗選取7日齡、2月、3月、4月、5月、6月、9月、18月齡晉嵐絨山羊公羊各4只,以其睪丸、附睪及18月齡山羊的精子為研究對象,利用QRT-PCR技術、Western-blot技術對山羊睪丸和附睪組織中Lcn5基因mRNA和Lcn5蛋白的相對表達量進行分析。利用免疫組化、免疫熒光技術對Lcn5在山羊睪丸、附睪及精子進行定位分析。主要結果如下：(1) QRT-PCR結果表明,Lcn5在山羊體內廣泛表達,生殖器官中表達量較高,其余器官中表達量均極低,且在生殖器官中的表達量為附睪頭附睪尾附睪體睪丸。不同發(fā)育階段附睪頭、附睪體、附睪尾及睪丸中Lcn5 mRNA的表達量在5月齡前隨著月齡的升高逐漸升高,5月齡時最高,隨后表達量又逐漸降低。(2) Western-blot結果顯示,Lcn5在不同時期的山羊睪丸和附睪中的表達量為：附睪頭附睪尾附睪體睪丸,不同月齡的山羊附睪頭、附睪體、附睪尾和睪丸中Lcn5蛋白的表達趨勢與Lcn5 mRNA表達趨勢一致。(3)免疫組化及免疫熒光結果顯示：Lcn5在山羊睪丸和附睪中均有表達,在睪丸精原細胞和精子細胞中微量表達,在附睪管假復層柱狀上皮細胞、纖毛細胞及管腔液中檢測到強陽性信號。精子的免疫熒光結果顯示Lcn5主要定位于精子的頂體及尾部中段。不同月齡山羊睪丸和附睪的免疫組化光密度結果分析與mRNA和Western-blot結果相一致,推測Lcn5可能在精子發(fā)生、成熟、獲能和精卵識別等方面發(fā)揮重要的作用。
[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.

【學位授予單位】：山西農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S827

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