MacroH2A deletion causes neuronal developmental deficits and autism-like behavior in mice

Purpose:To further understand how MacroH2A regulates the proliferation of neural progenitor cells, RNA-seq was used to analyze genome-wide changes in the cerebral cortex and littermate wild type of E13.5 MacroH2A knockout mice Methods: Total RNA was extracted from wild-type E13.5 brain tissue and knock-out E13.5 brain tissue. Specifically, the Agilent 2100 Bioanalyzer was used for quality control and quantification. Total RNA was then converted to cDNA and combined with the library. Illumina HiSeq 2500 platform in Annoroad Genomics uses RNA sequencing analysis Results: Approximately one thousand transcripts showed differential expression between the wild-type(WT) and knock-out mice brain cortex, with a fold change >=1.5 and p value <0.05.Geneontology analysis of the up-regulated genes showed obvious enrichment of biological processes related to development and proliferation.The down-regulated genes exhibited enrichment of biological processes related to the negative regulation of cell proliferation and developmental process. These results reflected human MacroH2A plays a role in brain neuron development. Total mRNA profiles of E13.5 wild type (WT) and knock-out (KO) mice were generated by deep sequencing using Illumina HiSeq 2500.研究目的：为深入阐明人源MacroH2A对神经祖细胞增殖的调控机制，本研究采用RNA测序（RNA-seq）技术，分析E13.5天MacroH2A基因敲除小鼠与其同窝野生型小鼠大脑皮层的全基因组转录变化。 实验方法：分别提取E13.5天野生型（wild-type, WT）与基因敲除（knock-out, KO）小鼠的脑组织总RNA，具体操作如下：首先使用安捷伦2100生物分析仪（Agilent 2100 Bioanalyzer）对总RNA进行质量控制与定量；随后将总RNA反转录为互补脱氧核糖核酸（cDNA）并构建测序文库；最后依托安诺优达基因科技（Annoroad Genomics）的Illumina HiSeq 2500平台完成RNA测序分析。 实验结果：野生型（WT）与基因敲除（KO）小鼠大脑皮层中，约有1000个转录本出现差异表达（差异倍数≥1.5，p值<0.05）。对上调基因进行基因本体（Gene Ontology, GO）富集分析发现，其显著富集于发育与增殖相关的生物学过程；下调基因则显著富集于细胞增殖负调控及发育过程相关的生物学过程。上述结果表明，人源MacroH2A在脑神经元发育过程中发挥重要调控作用。本研究通过Illumina HiSeq 2500平台进行深度测序，获取了E13.5天野生型与基因敲除小鼠脑组织的全mRNA表达谱。