梅花鹿早期妊娠不同时期血液转录组分析

Transcriptomic Analysis of Blood in Early Pregnancy at Different Time Points in Sika Deer

【目的】通过分析梅花鹿早期妊娠不同时期的血液转录组数据,筛选参与梅花鹿早期妊娠的关键基因。【方法】分别采集人工授精后0、7、15和20 d的雌性梅花鹿血液样品,利用Illumina HiSeq TM 3000C测序平台进行转录组分析。对不同时期的血液样本进行主成分分析和组间表达相关性分析,鉴定梅花鹿早期妊娠的关键阶段,利用DESeq筛选各时期的差异表达基因(DEGs),并进行GO和KEGG功能分析以及蛋白互作网络构建。【结果】转录组测序结果显示,妊娠第15天时梅花鹿血液出现了转录高峰,比较组D7 vs D0、D15 vs D7、D20 vs D15分别筛选到181、996、265个DEGs。其中比较组D7 vs D0共筛选到ERAS、MET、KDR等7个关键基因。综合GO和KEGG功能富集结果显示,基因主要富集在细胞对刺激的响应、信号转导等生物过程,以及PI3K-Akt信号通路、MAPK信号通路等通路中;比较组D15 vs D7共筛选到HRAS、HIF 1A、MAPK 1、JAK 1等10个关键基因,主要富集在免疫系统进程、免疫反应、趋化因子信号通路、C型凝集素受体信号通路等信号通路中;比较组D20 vs D15共筛选到MAPK 1、MMP 2等4个关键基因,主要富集在雌激素信号通路和松弛素信号通路中。【结论】本研究分析了梅花鹿早期妊娠关键时间点血液转录组的动态表达,筛选到KDR、HRAS、MAPK 1、JAK 1等关键基因,为梅花鹿早期妊娠诊断标志物的挖掘提供参考。

【Objective】The objective of this study was to analyze the blood transcriptome data at different stages of early pregnancy in sika deer and identify key genes involved in early pregnancy.【Method】Blood samples from female sika deer were collected at 0,7,15,and 20 days after artificial insemination.Transcriptome sequencing was performed using the Illumina HiSeq TM 3000C sequencing system to select differentially expressed genes.Principal component analysis and inter-group expression correlation analysis were conducted on blood samples at different time points to identify key stages of early pregnancy in sika deer.DESeq was used to screen differentially expressed genes(DEGs)at each time period,followed by GO,KEGG,and protein-protein interaction network analysis.【Result】The transcriptome sequencing results revealed a transcriptional peak in sika deer blood on day 15 of pregnancy.Comparing the groups D7 vs D0,D15 vs D7,and D20 vs D15,181,996,and 265 DEGs were identified,respectively.Among them,a total of 7 key genes,including ERAS,MET and KDR,were screened in comparison group D7 vs D0.Comprehensive analysis of GO and KEGG enrichment results indicated that these genes were mainly enriched in biological processes such as cellular response to stimuli and signal transduction,as well as pathways including the PI3K-Akt signaling pathway,and MAPK signaling pathway.A total of 10 key genes,including HRAS,HIF 1A,MAPK 1,and JAK 1,were screened in comparison group D15 vs D7.The genes were mainly enriched in immune system processes,immune response,chemokine signaling pathway,and C-type lectin receptor signaling pathway.A total of 4 key genes,including MAPK 1 and MMP 2,were identified in comparison group D20 vs D15.The genes were mainly enriched in the estrogen signaling pathway and relaxin signaling pathway.【Conclusion】The dynamic expression of blood transcriptome at key time points during early pregnancy in Sika deer was analyzed,and key genes such as KDR,HRAS,MAPK 1,and JAK 1 were screened,providing reference for the exploration of diagnostic markers for early pregnancy in sika deer.