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疼痛应答中背根神经节可变剪接的鉴定与分析 预览

Identification and Analysis of the Alternative Splicing in Dorsal Root Ganglion in the Pain Response
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摘要 目的鉴定和分析完全佛氏佐剂诱导绵羊炎性疼痛后背根神经节(DRG)发生的可变剪接事件和差异剪接基因,从转录后调控水平揭示大型动物对慢性疼痛应答调控的分子机制。方法实验组采用完全弗氏佐剂建立炎性疼痛模型,对照组则不建立疼痛模型,对2组绵羊的DRG组织进行转录组测序,利用Tophat软件鉴定和分析绵羊DRG中发生可变剪接事件和可变剪接差异基因,并对差异基因进行GO功能富集分析。结果测序结果显示,对照组和实验组DRG中分别鉴定出8 527和6 212个可变剪接事件,其中内含子保留为最多事件,鉴定出360个显著的差异剪接基因(P<0.05)。GO功能富集分析显示,差异剪接基因显著富集到电压门控离子通道、钠离子结合和钠离子运输相关及钙离子运输、钙离子通道、调控Ras蛋白信号转导的基因、调控Ras-GTP酶活性等功能上。其中HCN,CLCN3,KCNC4,CACNA2D1,TRPC4,CACNA2D1,ACAP2,GIT2,TRIO基因的可变剪接模式存在显著差异。结论可变剪接在疼痛应答调控中发挥重要作用,特别是电压门控离子通道相关基因可能通过可变剪接模式的改变来参与疼痛应答。 Objective To identify and analyze alternative splicing event (AS) and differential splicing gene (DSG) in the dorsal root ganglion of sheep with inflammatory pain induced by complete Freund's adjuvant, the regulatory mechanism of chronic pain response was revealed at the level of post-transcriptional regulation in large animals. Methods Pain model was established with complete freund's adjuvant inducing in the experimental group , while no pain model was established in the control group.The transcriptomic profiling of the dorsal root ganglion in sheep was sequenced,and the ASand DSG were identified and analyzed using Tophat software. The GO function enrichment analysis were performed for DSG. Results The results showed that 8 527 and 6 212 alternative splicing events were identified in genome of dorsal root ganglion in the experimental group and control group, respectively, and the proportion of IntronR was the highest. The GO analysis showed that differentially spliced genes were significantly enriched in voltage-gate ion channels,sodium ion binding and sodium ion transport,calcium ion transport,calcium ion channels, regulating the Ras-GTP enzyme activity and the Ras protein signal transduction related genes made changed significantly in alternative splicing events,including HCN, CLCN3, KCNC4, CACNA2D1, TRPC4, CACNA2D1, ACAP2, GIT2, TRIO and so on. Conclusion These results suggest that alternative splicing plays an important role in the pain response regulation, especially voltage gated ion channel related genes may participate in pain response through the change of alternative splicing mode.
作者 邓秀玲 王海生 叶纪诚 张建宇 孙计桃 苑红 DENG Xiuling;WANG Haisheng;YE Jicheng;ZHANG Jianyu;SUNJitao;YUAN Hong(College of Basic Medicine, Inner Mongolia Medical University, Hohhot Inner Mongolia 010110, China)
出处 《中国继续医学教育》 2019年第23期149-151,共3页 China Continuing Medical Education
关键词 炎性疼痛 背根神经节 转录组测序 可变剪接 离子通道 疼痛应答 inflammatory pain dorsal root ganglion RNA sequencing alternative splicing ion channel pain response
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