藏猪、川乡黑猪妊娠期子宫体繁殖功能相关lncRNA鉴定和功能预测

Identification and Function Prediction of lncRNA Related to Reproductive Function of Uterine Body in Tibetan and Chuanxiang Black Pigs During Pregnancy

[目的]探讨长链非编码RNA(long non-coding RNA,lncRNA)在藏猪、川乡黑猪繁殖过程中的调控机理,筛选相关lncRNA。[方法]采集具有不同产仔数的藏猪和川乡黑猪妊娠期的子宫体组织作为样本,建立特异性猪子宫mRNA文库,利用Illumina PE150 HiSeq平台进行测序,测序结果经过质控、比对与拼接后,筛选出差异表达lncRNA并预测其靶基因,对靶基因进行GO功能和KEGG通路富集分析。随机选取7个差异表达lncRNAs进行实时荧光定量PCR验证。[结果]在藏猪和川乡黑猪子宫体中存在32个共同差异表达lncRNAs,共得到168个靶基因。GO功能分析显示,不同繁殖能力的样本共同差异表达lncRNA的靶基因主要富集于分子代谢、转录活性调节、细胞增殖调节、子宫体修饰、微量元素代谢等条目中;KEGG通路分析显示,共同差异表达lncRNA的靶基因主要富集在糖酵解过程、卵母细胞分裂、细胞寿命调节、Ras信号通路、RIG-Ⅰ信号通路、FoxO信号通路、HIF-1信号通路、调控稳态过程等。实时荧光定量PCR结果显示,7个差异表达lncRNAs的表达与转录组测序结果一致。[结论]本研究在藏猪和川乡黑猪子宫体中发现了32个可能通过调控潜在靶基因参与母猪妊娠过程的lncRNAs,为进一步研究lncRNA在母猪繁殖过程中的调节机制提供了参考依据。这些lncRNA可作为新的繁殖性状生物标记用于猪的育种过程中。

[Objective]This study was aimed to investigate the regulatory mechanism of long non-coding RNA(lncRNA)in reproduction of Tibetan and Chuanxiang Black pigs,and screen the related lncRNA.[Method]The pregnant uterine body samples from Tibetan and Chuangxiang Black pigs with different litter sizes were collected,a specific porcine uterine body mRNA library was established,and sequencing was performed on the Illumina PE150 HiSeq platform.After quality control,alignment and splicing,the differentially expressed lncRNA was screened and its target gene was predicted.GO function and KEGG pathway enrichment analysis were performed on the target genes.In addition,7 differentially expressed lncRNAs were randomly selected for Real-time quantitative PCR verification.[Result]There were 32 co-differentially expressed lncRNAs in uterine body of Tibetan and Chuanxiang Black pigs,and a total of 168 target genes were obtained.GO function analysis showed that the target genes of co-differentially expressed lncRNAs in samples with different reproductive ability were mainly enriched in items such as molecular metabolism,transcriptional activity regulation,cell proliferation regulation,uterine body modification,trace element metabolism,etc.KEGG pathway analysis showed that the target genes of co-differentially expressed lncRNAs were mainly enriched in glycolysis,oocyte division,cell life regulation,Ras signaling pathway,RIG-Ⅰsignaling pathway,FoxO signaling pathway,HIF-1 signaling pathway,regulation of homeostasis process,etc.Real-time quantitative PCR results showed that the expression of 7 differentially expressed lncRNAs were consistent with RNA-Seq results.[Conclusion]This study found 32 lncRNAs in uterine body of Tibetan and Chuanxiang Black pigs that might be involve in the pregnancy process of sows by regulating potential target genes,so as to provide a reference basis for further research on the regulatory mechanism of lncRNA in sow reproduction.These lncRNAs could be used as new reproductive trait biomarkers for the breeding process of pigs.