川中黑山羊和雷州山羊大小卵泡转录组学分析

Comparative Transcriptome Analysis Between Large and Small Folliclesin Chuanzhong Black and Leizhou Goats

试验旨在通过比较两个品种山羊大小卵泡的mRNA表达图谱来挖掘影响山羊卵泡发育的基因,为进一步阐述山羊卵泡发育机制提供数据基础。使用氯前列醇钠对川中黑山羊和雷州山羊进行同期发情处理,屠宰后采集卵巢并在体视显微镜下分离单个卵泡(大卵泡>6 mm;小卵泡<3 mm),提取卵泡组织总RNA进行RNA-seq,利用生物信息学方法检测mRNA表达谱,筛选差异基因,并对其进行GO功能、KEGG通路富集分析。结果显示,川中黑山羊大小卵泡差异基因共4451个,雷州山羊2355个,二者共有差异基因1771个。分析筛选出两个品种共有(INHBA、INHA、CYP19A1、KITLG、LHCGR和STAR)及各自特有(IGFBP6、BMP6和BMPR2)的已报道与卵泡发育相关的基因,此外还筛选出可能与这两种山羊繁殖性能相关的基因(GADD45B、TC2N和MSMO1)。GO功能分析显示,两个品种共有差异基因主要与各种物质的跨膜转运活性有关;KEGG通路分析显示,类固醇生成、细胞因子受体相互作用和cAMP信号通路在两个品种中均存在显著变化。类固醇生成过程中细胞色素P450代谢差异可能是川中黑山羊高产仔数的一个潜在因素。本研究结果为进一步探究山羊卵泡发育的基因功能及不同山羊品种繁殖性能差异提供参考。

This study was aimed to excavate the genes affecting follicular development with the mRNA transcriptome difference between large and small follicles in two different goats,in order to provide a reference for further elucidating the mechanism of goat follicular development.Synchronization of estrus was carried out on the Chuanzhong Black and Leizhou goats with cloprostenol sodium.The single follicle(large follicle>6 mm,small follicle<3 mm)was isolated from ovaries which were collected from estrus goats mentioned above with the stereomicroscope.The total RNA was extracted from follicle tissues for RNA-seq,the profiles of mRNA expression were calculated with bioinformatics and differentially expressed genes(DEGs)were performed in GO function and KEGG pathway enrichment.The results showed that 4451 DEmRNAs were obtained from different follicles in Chuanzhong Black goats,and 2335 DEmRNAs were obtained from Leizhou goats,two goat breeds 1771 genes.The common(INHBA,INHA,CYP19A1,KITLG,LHCGR and STAR)and unique(IGFBP6,BMP6 and BMPR2)genes related to follicular development were screened out.In addition,the new genes(GADD45B,TC2N and MSMO1)potentially affecting the reproductive performance in two goat breeds were also identified.GO function analysis showed that the DEGs in two goat breeds were mainly related to the transmembrane transport activity.KEGG pathway analysis revealed significant changes in steroidogenesis,cytokine-cytokine receptor interaction and cAMP signaling in both goat breeds.It was also found that the difference of cytochrome P450 metabolism in steroidogenesis when follicle developed could be a potential factor in the high litter size of Chuanzhong Black goats.The results provided a reference for further exploring the gene functions of follicle development and the research in reproductive performance differences of different goat breeds.