基于高通量测序的山羊卵巢基质、卵泡转录组分析

    Transcriptome analysis of goat ovaries and follicles based on high-throughput sequencing

    • 摘要:
      目的  比较山羊Capra hircus卵巢基质、大卵泡和小卵泡间的mRNA表达图谱,为探究卵泡发育机制提供一定的研究基础。
      方法  采用高通量测序技术对发情期川中黑山羊的卵巢基质、大卵泡和小卵泡进行转录组测序,利用生物信息学方法检测mRNA表达谱,筛选差异基因,对其进行GO和KEGG通路分析,最后随机抽取5个差异基因进行荧光定量PCR验证。
      结果  卵巢基质vs大卵泡、卵巢基质vs小卵泡和小卵泡vs大卵泡的差异表达基因分别为524、180和403个。筛选出INHATNFRSF19等15个与卵泡发育相关的基因,其主要富集在内质网蛋白质加工、类固醇生物合成、卵母细胞减数分裂等信号通路。通过qRT-PCR验证,定量结果与测序结果基本一致。
      结论  川中黑山羊卵巢基质、大卵泡和小卵泡的基因表达模式不同,小卵泡与卵巢基质的基因表达模式更相近。

       

      Abstract:
      Objective  To compare the expression profiles of mRNA among ovarian stroma, large follicles and small follicles of goat (Capra hircus), and lay a foundation for exploring the mechanism of follicular development.
      Method  High-throughput sequencing technology was used to sequence the transcriptome of ovarian stroma, large follicles and small follicles in Chuanzhong black goats during estrus. The expression profiles of mRNA were detected by bioinformatics, and the differentially expressed genes were screened out. The GO and KEGG pathways of differentially expressed genes were analyzed. Finally, five differentially expressed genes were randomly selected and were verified by fluorescence quantitative PCR.
      Result  The differentially expressed genes of ovarian stroma vs large follicles, ovarian stroma vs small follicles and small follicles vs large follicles were 524, 180 and 403, respectively. Fifteen genes related to follicular development, such as INHA and TNFRSF19, were screened. They were mainly involved in endoplasmic reticulum protein processing, steroid biosynthesis, oocyte meiosis signaling pathway and so on. Quantitative results were basically consistent with sequencing results verified by qRT-PCR.
      Conclusion  The gene expression profiles of ovarian stroma, large follicles and small follicles in Chuanzhong black goats are different, and the gene expression patterns of small follicles and ovarian stroma are relative more similar.

       

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