基于垂体转录组测序技术的北极狐体型变异相关基因筛选OA
Screening of genes related to body size variation in Alopex lagopus based on pituitary transcriptome sequencing technology
为筛选与北极狐(Alopex lagopus)体型变异相关的基因,试验通过对 300 只 9 月龄、健康、雄性北极狐进行体重和体长测定,选择体型较大(大体型组)和较小(小体型组)个体各 3 只,利用 Illumina NovaSeq 6000 平台对其垂体组织进行转录组测序,根据测序结果筛选差异表达基因(DEGs),通过对 DEGs 进行基因本体论(GO)功能与京都基因与基因组百科全书(KEGG)通路富集分析及蛋白质-蛋白质相互作用(PPI)网络分析筛选与北极狐体型变异相关的基因,并采用实时荧光定量PCR 方法对候选基因进行验证.结果表明:共筛选到413 个DEGs,其中大体型组有248 个DEGs为上调表达基因,165 个 DEGs 为下调表达基因;这些 DEGs 富集于 65 个 GO 条目上,包括单一生物体过程、细胞成分的组织或生物发生、细胞膜组分、信号转导活性等与体型变异相关的生物学功能,涉及到的基因有 IHH、GHSR、GREM1 和SSTR5;同时富集于217 条KEGG 通路上,包括神经活性配体-受体相互作用(GHSR、SSTR5 基因)、cAMP 信号通路(GHSR、SSTR5 基因)、刺猬信号通路(IHH 基因)和 TGF-β 信号通路(GREM1 基因)等与体型变异相关的信号通路;GHSR、SSTR5 和 SST 等蛋白位于 PPI 网络的核心节点;实时荧光定量 PCR 验证发现,与小体型组比较,大体型组垂体中 IHH和 GHSR 基因的相对表达量极显著升高(P<0.01),GREM1 和 SSTR5 基因的相对表达量极显著降低(P<0.01),趋势与转录组测序结果相符.说明北极狐垂体中 IHH、GHSR、GREM1和 SSTR5等基因可能与其体型变异有关.
In order to screen the genes related to body size variation in Alopex lagopus,three individuals with larger body size(large body size group)and three individuals with smaller body size(small body size group)were selected by measuring the weight and length of 300 healthy male Alopex lagopus aged 9 months.The pituitary tissues of these individuals were subjected to transcriptome sequencing using the Illumina NovaSeq 6000 platform.Differentially expressed genes(DEGs)were identified based on the sequencing results.GO functional and KEGG pathway enrichment analysis,as well as PPI network analysis,were conducted on the DEGs to screen the genes related to body size variation in Alopex lagopus.Real-time fluorescence quantitative PCR was used to verify the candidate genes.The results showed that a total of 413 DEGs were identified,including 248 expression up-regulated and 165 expression down-regulated DEGs in the large body size group.These DEGs were enriched in 65 GO terms,including single biological processes,cell component organization and biogenesis,cell membrane components,signal transduction activities,which were related to body size variation.The genes involved included IHH,GHSR,GREM1,and SSTR5.They were also enriched in 217 KEGG pathways,including neuroactive ligand-receptor interaction(GHSR,SSTR5 genes),cAMP signaling pathway(GHSR,SSTR5 genes),hedgehog signaling pathway(IHH gene),and TGF-β signaling pathway(GREM1 gene),which were related to body size variation.GHSR,SSTR5,and SST genes were located at the core nodes of the PPI network.Real-time fluorescence quantitative PCR verification showed that the relative expression levels of IHH and GHSR genes were significantly higher than those in the small body size group(P<0.01),while the relative expression levels of GREM1 and SSTR5 genes in the large body size group were significantly lower than those in the small body size group(P<0.01),which was consistent with the results of transcriptome sequencing.The results indicated that genes such as IHH,GHSR,GREM1,and SSTR5 in the pituitary gland of Alopex lagopus might be related to their body size variation.
刘庆;李佳鹏;王晨阳;王雨萌;马泽芳;周李生;崔凯
青岛农业大学 动物科技学院,山东 青岛 266109青岛农业大学 动物科技学院,山东 青岛 266109青岛农业大学 动物科技学院,山东 青岛 266109青岛农业大学 动物科技学院,山东 青岛 266109青岛农业大学 动物科技学院,山东 青岛 266109青岛农业大学 动物科技学院,山东 青岛 266109青岛农业大学 动物科技学院,山东 青岛 266109
农业科技
北极狐垂体生长发育转录组测序差异表达基因
Alopex lagopuspituitarygrowth and developmenttranscriptome sequencingdifferentially expressed genes
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