【摘要】 旨在通过对崇仁麻鸡背部皮肤毛囊组织的转录组学分析，筛选关键的差异表达基因和信号通路，探讨鸡皮肤毛囊性状的分子调控机制。采用TRIzol法提取皮肤毛囊组织总RNA,通过Illumina平台进行转录组高通量测序(RNA-Seq)。最后采用实时荧光定量的方法对转录组测序进行验证。研究结果如下：共筛选出了3 856个差异表达基因，在公鸡(BGB vs YGB)比较组合中筛选出了971个差异表达基因，其中274个表达上调，697个表达下调。在母鸡(BMB vs YMB)比较组合中筛选出了3 529个差异表达基因，其中1 477个上调，2 052个下调。在公鸡、母鸡共同的差异表达基因中筛选出了3个与皮肤毛囊性状有关的高表达基因KRT75、KRT6A和KRT14。在GO功能显著富集中，BGB vs YGB比较组合富集到了516个GO term, BMB vs YMB比较组合富集到了1 020个GO term,基因主要集中参与细胞黏附等活动。显著富集了多条KEGG通路，在BGB vs YGB比较组合中富集了8条显著的KEGG通路，在BMB vs YMB比较组合中富集了20条显著的KEGG通路，筛选出了3条关键通路Wnt信号传导途径、神经活性配体-受体调控通路和TGF-β调控通路。荧光定量结果显示，随机选取的5个差异基因在皮肤毛囊中的表达与RNA-Seq走向趋势一致。综上所述，筛选的KRT75、KRT6A、KRT14基因可能影响鸡皮肤毛囊性状；神经活性配体-受体调控通路、Wnt信号通路和TGF-β信号通路均是调控鸡皮肤毛囊性状的重要通路。更多还原

【Abstract】 The aim was to screen key differentially expressed genes and signaling pathways and explore the molecular regulatory mechanisms of chicken skin follicle traits through transcriptomic analysis of the dorsal skin follicle tissues of Chongren partridge chicken.Total RNA of skin hair follicle tissue was extracted by TRIzol method, and transcriptome high-throughput sequencing(RNA-Seq)was performed by Illumina platform.Finally, the transcriptome sequencing was validated by Real-time fluorescence quantitative PCR.The results were as follows: A total of 3 856 differentially expressed genes were screened, and 971 differentially expressed genes were screened in the cockerel(BGB vs YGB)comparative combinations, of these, 274 up-regulated and 697 down-regulated.There were 3 529 differentially expressed genes in comparative combinations of hen(BMB vs YMB),of these, 1 477 up-regulated and 2 052 down-regulated.Three highly expressed genes, KRT75,KRT6A,and KRT14,associated with skin follicle traits, were screened among the common differentially expressed genes in roosters and hens.In the significant enrichment of GO functions, 516 GO terms were enriched in the BGB vs YGB and 1 020 GO terms were enriched in the BMB vs YMB.Genes were mainly focused on activities involved in cell adhesion and cell activation.Significantly enriched multiple KEGG pathways, with 8 significant KEGG pathways enriched in the BGB vs YGB and 20 significant KEGG pathways enriched in the BMB vs YMB.Three key pathways, Wnt signaling pathway, neuroactive ligand-receptor regulatory pathway and TGF-β regulatory pathway, were screened.Fluorescence quantification showed that the expression of five randomly selected differential genes in skin hair follicles was consistent with the trend towards RNA-Seq.In summary, the screened KRT75,KRT6A,and KRT14 genes may affect chicken skin follicle traits; the neuroactive ligand-receptor regulatory pathway, the Wnt signaling pathway, and the TGF-β signaling pathway are important pathways that regulate follicular traits in chicken skin.更多还原