蒙古马睾丸发育及精子生成相关基因的筛选与分析

发布时间：2018-04-06 06:34

本文选题：蒙古马　切入点：睾丸　出处：《内蒙古农业大学》2017年硕士论文

【摘要】：蒙古马是世界上最古老的马品种之一。具有良好的繁殖能力和独特的遗传基因。通过测量马睾丸可有效预测种公马的繁殖能力。睾丸发育是雄性哺乳动物重要的生理过程。产生精子和分泌雄性激素是睾丸的基本功能。为了从基因水平了解性成熟前后蒙古马睾丸发育及精子生成的调控机制,本实验采用新一代高通量测序技术,对蒙古马性成熟前后两个发育阶段的睾丸组织建立转录组文库进行测序、对结果进行可变剪切分析及差异表达基因分析;对差异表达基因进行GO功能富集分析、KEGG Pathway分析、转录因子分析,找到睾丸发育及精子生成相关GO功能条目、KEGG通路及相关基因。最后应用实时荧光定量PCR方法检测蒙古马睾丸中TSSK2、SPATA6、CDC20、MAD2L1基因的mRNA表达量在性成熟前和性成熟后的变化。本研究对于选择优良种用公马具有重要意义。揭示公马睾丸发育及精子生成的分子机制,并获得以下研究结果:(1)经过对蒙古马性成熟前后期进行转录组分析,得到24878个差异表达基因,其中上调基因16237个,下调基因8641个。这些差异基因被注释到10019条GO功能条目上。主要富集在精子形成(spermatogenesis)、雄性配子的一代(male gamete generation)、精细胞的发育(spermatid development)等与睾丸发育及精子生成紧密相关的生物学过程。(2)对差异表达基因进行KEGG通路分析发现:与睾丸发育及精子生成紧密相关的通路有卵母细胞减数分裂(Oocyte meiosis)、黄体酮调节的卵母细胞减数分裂(Progesterone-mediated oocyte maturation)等通路。(3)采用实时荧光定量PCR技术对TSSK2基因、SPATA6基因、CDC20基因及MAD2L1基因进行验证,结果显示以上4个基因在性成熟后睾丸中的表达量比性成熟前睾丸中的表达量高2倍以上,且差异显著。证明了转录组测序结果的准确性。
[Abstract]:Mongolian horse is one of the oldest horse breeds in the world.Has good reproduction ability and unique genetic genes.By measuring the testis of the horse, the breeding ability of the male horse can be predicted effectively.Testicular development is an important physiological process in male mammals.The production of sperm and the secretion of testosterone are the basic functions of the testis.In order to understand the regulatory mechanism of testis development and spermatogenesis in Mongolian horses before and after sexual maturation at the gene level, a new generation of high-throughput sequencing technique was used.The transcriptional library of testis in two stages of sexual maturation was sequenced, the results were analyzed by variable shear analysis and differential expression gene analysis, and the differentially expressed genes were analyzed by go functional enrichment analysis and KEGG Pathway analysis.Transcription factor analysis revealed KEGG pathway and related genes related to testicular development and spermatogenesis.The changes of mRNA expression of TSSK2SPATA6SPATA6CDC20 / MAD2L1 gene in the testis of Mongolian horses before and after sexual maturation were detected by real-time fluorescence quantitative PCR.This study is of great significance for the selection of a good breed of male horse.The molecular mechanism of testicular development and spermatogenesis in male horses was revealed, and the following results were obtained: 1) 24878 differentially expressed genes were obtained by transcriptome analysis before and after sexual maturation of Mongolian horses, including 16237 up-regulated genes and 8641 down-regulated genes.These differential genes were annotated to 10019 go functional entries.The differential expression genes were mainly enriched in spermatogenesis, male gamete generation gamete generation, spermatocyte development and spermatid development and other biological processes closely related to testicular development and spermatogenesis. The results of KEGG pathway analysis of differentially expressed genes were as follows:The pathways closely related to development and spermatogenesis include oocyte meiosism, progesterone-mediated oocyte maturation.3.) the TSSK2 gene, SPATA6 gene, CDC20 gene and MAD2L1 gene were verified by real-time fluorescence quantitative PCR.The results showed that the expression of the above four genes in the testis after sexual maturation was more than 2 times higher than that in the testis before sexual maturation, and the difference was significant.The results of transcriptome sequencing were proved to be accurate.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S821.81

【参考文献】