【摘要】 目的利用Dicer1条件敲除小鼠,通过RNA-seq技术在转录组水平分析microRNAs(miRNAs)对哺乳动物大脑早期发育的影响。方法构建D6-Cre-Dicer1条件敲除小鼠,实验分为对照组(Ctrl)和条件敲除组(cKO)。苏木精-伊红和免疫荧光染色比较敲除前后组织形态差异;选取E14.5 D6-Cre介导的Dicer1条件性敲除的小鼠背侧皮质,提取总RNA,利用RNA-seq技术分析Dicer1敲除前后转录水平差异。结果与对照组相比,条件敲除组1个月左右致死,P3时皮层神经元发育异常,皮层和海马结构部分缺失;通过RNA-seq分析,共筛选出差异表达基因45个,上调基因11个,下调基因34个(P<0.05)。差异基因富集于NF-κB信号通路、Wnt信号通路、MAPK-ERK信号通路、JAK-STAT信号通路和DNA损伤应激通路等。结论在发育早期E14.5时Dicer1参与调节多条信号通路,影响部分皮层层次标志物的表达,抑制lncRNA Xist的表达,调节皮质发育。更多还原

【Abstract】 Objective To explore the role of microRNAs(miRNAs) during early neurodevelopment in mammals with Dicer1 conditional knockout(cKO)mouse by RNA-seq. Methods The animals were divided into Ctrl group and cKO group. HE and immunofluorescence staining were used to observe the histological morphology; Extract total RNA of E14.5 D6-Cre-Dicer1 mice dorsal cortex, using RNA-seq technique to analyze differences at transcriptional level. Results Animals in cKO groups died within a month or so, abnormal development of cortical and hippocampal was recorded at P3. In cKO group transcription expression was different significantly, 45 differentially expressed genes were screened, including 11 up-regulated and 34 down-regulated genes(P<0.05)mainly enriched in Wnt signaling pathway, DNA damage response signaling pathway and others. Conclusions At E14.5 cortex, Dicer1 is involved in the regulation of multiple signaling pathways, affecting partial cortical laminar genes expres-sion, inhibiting the expression of lncRNA Xist and regulating cortical development.更多还原