本文選題：北京油雞種公雞　切入點：繁殖力　出處：《中國農(nóng)業(yè)科學院》2015年碩士論文

【摘要】：中國地方雞種肉蛋品質(zhì)優(yōu)良,抗病能力強,但繁殖效率較低。進一步調(diào)查發(fā)現(xiàn),地方雞無精癥、弱精癥比例較高,而且精液量的減少常常伴隨著精子濃度降低,提高了地方雞生產(chǎn)中公母配比上升。增加了動物生產(chǎn)成本。研究發(fā)現(xiàn):雞精液品質(zhì)性狀為中等遺傳力性狀,較大程度上受到遺傳因素的影響。本研究目的是探究影響繁殖力的遺傳因素。蛋白質(zhì)作為基因表達的產(chǎn)物,直接調(diào)控性狀。本研究通過篩選低繁殖力公雞與正常公雞,比較睪丸發(fā)育學和形態(tài)學特征,檢測低繁殖力公雞與正常公雞睪丸組織蛋白表達量的差異,利用同位素標記相對和絕對定量(iTRAQ)技術及生物信息學分析,篩選與繁殖力相關的候選蛋白,并進行蛋白質(zhì)印跡法驗證,為進一步深入研究影響公雞繁殖力及睪丸發(fā)育分子遺傳機制提供理論基礎和潛在方向。主要研究內(nèi)容及結果如下:1.低繁殖力公雞挑選及睪丸特征比較。試驗動物為42周齡相同環(huán)境下飼養(yǎng)北京油雞公雞200只,通過精液品質(zhì)檢測和交配試驗挑選出差異顯著的低繁殖力與正常種公雞各25只。52周齡獲取睪丸組織,稱重,制作組織切片并觀測睪丸精細小管形態(tài),綜合比較低繁殖力公雞與正常公雞睪丸參數(shù)即生精上皮長,精細小管直徑、面積,約翰遜評分等。結果顯示低繁殖力組公雞睪丸重量、生精上皮長、精細小管直徑、面積及約翰遜評分等睪丸參數(shù)顯著低于正常組。根據(jù)試驗結果,挑選出3只低繁殖力公雞及3只正常同胞公雞,作為iTRAQ試驗樣本。2.睪丸組織差異表達蛋白篩選、分析及驗證。利用iTRAQ技術,對樣本睪丸組織總蛋白進行相對及絕對定量分析。對iTRAQ定量結果的分析,進行低繁殖力和正常個體及組間的睪丸組織蛋白表達相互比較。結果發(fā)現(xiàn),低繁殖力組上調(diào)倍數(shù)較大的蛋白為天冬氨酸氨基轉(zhuǎn)移酶(AspAT)、谷胱甘肽S轉(zhuǎn)移酶(GST)蛋白和組蛋白H2A(Histone H2A),下調(diào)倍數(shù)較大的蛋白有DNA損傷修復蛋白(DDB2)、T型中心粒蛋白(CENP-T)和G型蛋白酪氨酸磷酸酶(PTPRG)。KEGG Pathway富集分析發(fā)現(xiàn)差異表達蛋白顯著富集于嘌呤和嘧啶的代謝、精氨酸和脯氨酸代謝等信號通路。利用蛋白免疫印跡法(Western Blotting)驗證GST蛋白、AspAT蛋白和Histone H2A蛋白在低繁殖力組和正常組睪丸組織中表達情況,結果發(fā)現(xiàn)蛋白表達趨勢與iTRAQ技術檢測結果一致。綜合試驗2結果,DDB2、CENP-I、AspAT、GST、S100-A6和Histone H2A為調(diào)控精子發(fā)生的候選蛋白,其中AspAT、GST、DDB2和Histone H2A為重要候選蛋白。GnRH信號通路、嘌呤和嘧啶的代謝和蛋白質(zhì)的轉(zhuǎn)運等信號通路可以作為公雞繁殖力相關的重要候選調(diào)控通路。
[Abstract]:Chinese local chicken breeds have good quality and strong resistance to disease, but the breeding efficiency is low. Further investigation found that the proportion of azoospermia and asthenospermia in local chickens is higher, and the decrease of spermatozoa is often accompanied by a decrease in sperm concentration. The ratio of male and female increased in the production of local chickens, and the cost of animal production was increased. The results showed that the quality of chicken semen was a medium heritability trait. The aim of this study was to investigate the genetic factors that affect fecundity. Protein, as a product of gene expression, directly regulates traits. By comparing testicular development and morphological characteristics, the differences of testicular protein expression between low fecundity rooster and normal cock were detected, and relative and absolute quantitative iTRAQ techniques and bioinformatics analysis were used. Candidate proteins associated with fecundity were screened and verified by Western blotting. The main contents and results of this study are as follows: 1. Selection of low fecundity rooster and comparison of testicular characteristics. A total of 200 Beijing fowl roosters were raised in the same environment at 42 weeks of age. Testicular tissue was obtained by semen quality test and mating test in 25 male chickens of low fecundity and 25 normal cocks at the age of .52 weeks. Tissue sections were made and the morphology of testicular fine tubules was observed. The testicular parameters of low fecundity rooster and normal cock were compared, such as the length of spermatogenic epithelium, the diameter of fine tubules, the area and Johnson's score, etc. The results showed that the weight of testis, the length of spermatogenic epithelium and the diameter of fine tubules in the low fecundity group. The testicular parameters, such as area and Johnson's score, were significantly lower than those in the normal group. According to the results of the experiment, three low fecundity roosters and three normal sibling roosters were selected as samples of iTRAQ test. Analysis and verification. Relative and absolute quantitative analysis of total protein in testicular tissue was carried out by using iTRAQ technique. In the analysis of quantitative results of iTRAQ, the low fecundity and the expression of testicular tissue protein in normal individuals and groups were compared. In low fecundity group, aspartate aminotransferase (AST), glutathione S-transferase (glutathione S-transferase) and histone (H2A(Histone H2AN) were the most up-regulated proteins. The down-regulated proteins included DNA damage repair protein (DNA damage repair protein), CENP-T) and G type. The enrichment analysis of protein tyrosine phosphatase (PTPRG). KEGG Pathway showed that the differentially expressed proteins were significantly enriched in the metabolism of purine and pyrimidine. The expression of GST protein AspAT protein and Histone H2A protein in testis of low fecundity group and normal group was detected by Western blotting. The results showed that the trend of protein expression was the same as that of iTRAQ technique. The results of experiment 2 showed that DDB2CENP-IAspATT S100-A6 and Histone H2A were candidate proteins for regulating spermatogenesis, among which AspAT-GST-DDB2 and Histone H2A were important candidate proteins. The signaling pathways of purine and pyrimidine metabolism and protein transport can be used as important candidate regulatory pathways related to fecundity of cock.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S831

【引證文獻】

相關會議論文 前1條

1 孟麗娜;張英杰;劉月琴;;綿、山羊高繁殖力主效基因的研究進展[A];中國畜牧獸醫(yī)學會養(yǎng)羊?qū)W分會2012年全國養(yǎng)羊生產(chǎn)與學術研討會議論文集[C];2012年

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本文編號：1694579