基于转录组数据挖掘不同日龄滩羊毛囊发育关键基因  

Discovering Key Genes in Hair Follicle Development of Tan Sheep at Different Days of Age Based on Transcriptome Data

作　　者：马思佳 赵正伟 王锦 马丽娜[1,2] MA Sijia;ZHAO Zhengwei;WANG Jin;MA Lina(College of Animal Science and Technology,Ningxia University,Yinchuan 750021,China;Institute of Animal Science,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002,China;Institute of Animal Science of CAAS,Beijing 100193,China)

机构地区：[1]宁夏大学动物科技学院,银川750021 [2]宁夏农林科学院动物科学研究所,银川750002 [3]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《中国畜牧兽医》2024年第9期3726-3738,共13页China Animal Husbandry & Veterinary Medicine

基　　金：宁夏回族自治区重点研发项目(2023BCF01007)。

摘　　要：【目的】比较不同日龄滩羊皮肤组织毛囊形态变化,挖掘影响毛囊发育的关键信号通路和候选基因,对揭示滩羊毛囊发育的分子调控机制具有重要意义。【方法】选取出生后0、35和55日龄的滩羊各3只,采集皮肤组织样品,利用HE染色法进行组织学分析;利用Illumina NovaSeq 6000平台进行转录组测序,采用DESeq2软件进行差异表达基因(DEGs)分析,并对DEGs进行KEGG通路富集分析,筛选出与毛囊发育相关的基因。随机选择10个DEGs用实时荧光定量PCR验证转录组测序结果。【结果】HE染色结果显示,随着滩羊日龄的增加,初级毛囊数量显著或极显著减少(P<0.05;P<0.01),次级毛囊数量极显著增加(P<0.01),且次级毛囊逐渐密集分布在初级毛囊周围,形成了毛囊群结构。转录组测序结果显示,在35和0日龄比较组中共筛选到681个DEGs,其中368个上调,313个下调;在55和0日龄比较组中共筛选到852个DEGs,其中469个上调,383个下调;在55和35日龄比较组中共筛选到194个DEGs,其中99个上调,95个下调。3个比较组DEGs取并集得到1274个DEGs,可分成4个簇(K1~K4 cluster)。对各簇进行KEGG富集分析发现,这些基因显著富集在PI3K-Akt和MAPK等与毛囊形态发生相关的信号通路中,其中PDGFRA、PDGFRB、PDGFB、FGFR 4和NR 4 A 1基因在毛囊发育和分子调控机制中发挥重要作用。实时荧光定量PCR结果显示,所选10个DEGs的表达水平与转录组测序结果一致,说明测序结果可靠。【结论】本研究通过转录组测序分析挖掘到PDGFRA、PDGFRB、PDGFB、FGFR 4和NR 4 A 1是毛囊发育过程中的重要候选基因,为进一步揭示滩羊裘皮性状形成的分子机制提供了理论依据。【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.1274 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,FGFR 4 a

关 键 词：滩羊 毛囊发育 差异表达基因 转录组测序 

分 类 号：S813.3[农业科学—畜牧学]