羊毛固醇合成酶杂合敲除小鼠全脑转录组学分析

Whole-brain transcriptome analysis of wool sterol synthase heterozygous knockout mice

目的探讨羊毛固醇合成酶(LSS)基因功能异常及胆固醇水平变化对大脑功能分子水平的影响。方法通过RNA-Seq分析LSS(+/-)小鼠及野生型(WT)C57BL/6小鼠基因的表达,KEGG富集分析差异基因涉及的信号通路。用实时定量PCR以及Western blot法检测差异基因的表达。结果通过RNA-Seq分析,LSS(+/-)杂合敲除导致小鼠大脑中320个基因表达出现变化,其中上调基因145个,下调175个,KEGG富集分析差异基因涉及神经活化的受体与配体通路、河马信号途径及谷氨酸能突触途径。实时定量PCR分析结果显示CCKBR与Htr5b分别出现上调。Western blot结果显示CCKBR基因在LSS(+/-)小鼠全脑组织总蛋白中明显高表达。结论LSS(+/-)杂合敲除会影响小鼠大脑功能及行为。

Objective To investigate the molecular changes of lanosterol synthase(LSS)gene dysfunction and cholesterol levels that may affect brain function.Methods The differentially expressed genes in LSS knockout(+/-)mice and WT C57BL/6 mice were analyzed by RNA-SEQ,and the signal pathways involved in the differentially expressed genes were analyzed by KEGG enrichment.The differential genes were detected by real-time quantitative PCR and the protein expression of differential genes was detected by Western blot.Results By RNA-SEQ analysis,LSS(+/-)heterozygous knockout resulted in the difference of 320 genes in the mouse brain,including 145 up-regulated genes and 175 down-regulated genes.KEGG enrichment analysis showed that these differentially expressed genes involved in neural-activated receptor and ligand pathways,hippocampal signaling pathways and glutamate synaptic pathways.Real-time quantitative PCR showed that CCKBR and Htr5b were up-regulated,respectively.Western blot results showed that CCKBR gene was highly expressed in the total protein of all brain tissues of LSS knockout(+/-)mice.Conclusion LSS heterozygotic knockout may affect brain function and animal behavior.