不同产驹数阳原驴血液转录组分析

Transcriptome Analysis of Blood in Yangyuan Donkeys with Different Litter Size

【目的】筛选影响阳原驴产驹数的关键基因与相关通路,提高阳原驴繁殖性能。【方法】以健康产双驹和产单驹阳原驴静脉全血为样本进行转录组高通量测序,与参考基因组非洲野驴(Equus asinus(ass))比对后筛选差异表达基因,并对差异表达基因进行GO功能及KEGG通路富集分析,筛选与阳原驴产驹数相关的基因。随机挑选5个差异表达基因进行实时荧光定量PCR验证。【结果】与产单驹组相比,产双驹组共筛选出1336个差异表达基因,其中723个上调,613个下调。GO功能富集分析显示,差异表达基因主要在受体活性、信号转导等条目中富集;KEGG通路富集分析显示,卵巢胆固醇生成和雌激素信号通路在富集通路前20位,并筛选出IGF1、IGF1R、INSR和ADCY3共4个与阳原驴产驹数相关的候选基因。实时荧光定量PCR结果与转录组测序结果一致,证实转录组测序结果可靠。【结论】不同产驹数阳原驴差异表达基因富集在卵巢类固醇生成和雌激素信号通路,IGF1、IGF1R、INSR和ADCY3基因参与卵巢激素分泌且均在产双驹组中表达上调。研究结果可为进一步阐明阳原驴不同产驹数的分子机制提供参考。

【Objective】The purpose of this experiment was to screen the key genes and related pathways that affected the litter size of Yangyuan donkeys,and improve the reproductive performance of Yangyuan donkeys.【Method】The venous blood samples of healthy Yangyuan donkeys with twins and singleton were used for transcriptome high-throughput sequencing.After comparison with the reference genome of the African wild donkey(Equus asinus(ass)),the differentially expressed genes were screened,GO function and KEGG pathway enrichment analysis were performed on the differentially expressed genes to screen the genes related to the litter size of Yangyuan donkeys.Five differentially expressed genes were randomly selected,and Real-time quantitative PCR was used for expression quantitative verification.【Result】A total of 1336 differentially expressed genes were screened out between twins and singleton groups,among which 723 genes were upregulated,and 613 genes were downregulated.GO function enrichment analysis showed that the differentially expressed genes were mainly enriched in receptor activity,signal transduction and other terms.KEGG pathway enrichment analysis showed that ovarian steroidogenesis and estrogen signaling pathways were in top 20 enriched pathways,and a total of 4 candidate genes related to the litter size of Yangyuan donkeys were screened out,including IGF 1,IGF 1 R,INSR and ADCY 3.Real-time quantitative PCR results were consistent with the transcriptome sequencing results,and the transcriptome sequencing results were reliable.【Conclusion】The enrichment of differentially expressed genes in Yangyuan donkeys with different number of foals was found in ovarian steroidogenesis and estrogen signaling pathways,IGF 1,IGF 1 R,INSR and ADCY 3 genes were involved in ovarian hormone secretion and up-regulated in twins group.The results provided a reference for further clarifying the molecular mechanism of different litter sizes in Yangyuan donkeys.