摘 要：試驗旨在通過比較兩個品種山羊大小卵泡的mRNA表達圖譜來挖掘影響山羊卵泡發育的基因，為進一步闡述山羊卵泡發育機制提供數據基礎。使用氯前列醇鈉對川中黑山羊和雷州山羊進行同期發情處理，屠宰後採集卵巢並在體視顯微鏡下分離單個卵泡(大卵泡>6 mm；小卵泡<3 mm)，提取卵泡組織總RNA進行RNA-seq，利用生物信息學方法檢測mRNA表達譜，篩選差異基因，並對其進行GO功能、KEGG通路富集分析。結果顯示，川中黑山羊大小卵泡差異基因共4 451個，雷州山羊2 355個，二者共有差異基因1 771個。分析篩選出兩個品種共有(INHBA、INHA、CYP19A1、KITLG、LHCGR和STAR)及各自特有(IGFBP6、BMP6和BMPR2)的已報導與卵泡發育相關的基因，此外還篩選出可能與這兩種山羊繁殖性能相關的基因(GADD45B、TC2N和MSMO1)。GO功能分析顯示，兩個品種共有差異基因主要與各種物質的跨膜轉運活性有關；KEGG通路分析顯示，類固醇生成、細胞因子受體相互作用和cAMP信號通路在兩個品種中均存在顯著變化。類固醇生成過程中細胞色素P450代謝差異可能是川中黑山羊高產仔數的一個潛在因素。本研究結果為進一步探究山羊卵泡發育的基因功能及不同山羊品種繁殖性能差異提供參考。
關鍵詞：山羊; 卵泡; 轉錄組; 基因
Abstract: 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 4 451 DEmRNAs were obtained from different follicles in Chuanzhong Black goats, and 2 335 DEmRNAs were obtained from Leizhou goats, two goat breeds 1 771 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.
Key words: goats; follicle; transcriptome; gene
刊載於《中國畜牧獸醫》2020年第5期,P1459-1471
DOI: 10.16431/j.cnki.1671-7236.2020.05.020