Discovering Key Genes in Hair Follicle Development of Tan Sheep at Different Days of Age Based on Transcriptome Data
[Objective]By comparing the morphological changes of hair follicles of skin tissues in Tan sheep at different days of age,this study was aimed to explore key signaling pathways and candidate genes influencing hair follicle development,which was of great significance to reveal the molecular regulation mechanism of hair follicle development in Tan sheep.[Method]The skin tissue samples from Tan sheep at 0,35 and 55 days of age(n=3)were collected for histological analysis using HE staining method,respectively,RNA-Seq was conducted through Illumina NovaSeq 6000 platform,and differentially expressed genes(DEGs)was analyzed through DESeq2 software.KEGG pathway enrichment analysis was also performed on DEGs to further screen genes related to hair follicle development,and 10 DEGs were randomly selected for Real-time quantitative PCR validation of RNA-Seq results.[Result]HE staining results showed that with the days of age in Tan sheep increasing,the number of primary hair follicles was significantly or extremely significantly decreased(P＜0.05 or P＜0.01),while the number of secondary hair follicles was extremely significantly increased(P＜0.01),and follicle cluster structures were formed by secondary hair follicles which were gradually distributed around primary hair follicles.RNA-Seq results indicated that a total of 681 DEGs were screened in B35 vs B0 comparison group,including 368 up-regulated and 313 down-regulated,a total of 852 DEGs were screened in B55 vs B0 comparison group,including 469 up-regulated and 383 down-regulated,and a total of 194 DEGs were screened in B55 vs B35 comparison group,including 99 up-regulated and 95 down-regulated.1 274 DEGs were yielded from the intersection of 3 comparison groups of DEGs,which could divide into 4 clusters(K1-K4).After conducting KEGG enrichment analysis on each cluster,it was found that these genes had a significant enrichment in signaling pathways associated with hair follicle morphogenesis such as PI3K-Akt and MAPK.Among these genes,PDGFRA,PDGFRB,PDGFB,FGFR4 and NR4A1 played important roles in hair follicle development and molecular regulation mechanism.The results of Real-time quantitative PCR showed that the expression of 10 DEGs were consistent with RNA-Seq results,which indicated the reliability of the sequencing results.[Conclusion]This study identified that PDGFRA,PDGFRB,PDGFB,FGFR4 and NR4A1 were important candidate genes in the process of hair follicle development through RNA-Seq analysis,which provided a theoretical basis for further elucidation of the molecular mechanisms underlying the fur characters of Tan sheep.
Tan sheephair follicle developmentdifferentially expressed genesRNA-Seq
万方数据
维普
