Single cell RNAseq analysis in embryonic spinal cords to profile transcriptomes in control and Jmjd3cKO motor neurons.

To characterize Jmjd3 function in motor neuron development, we isolated GFP+ cells derived from Olig2 expressing progenitor cells that included motor neuron population and then employed single cell RNAseq. We found some novel motor neuron subtypes in addition to well-known motor columns, and specific genes in each subtype by unbiased and comprehensive analysis. Moreover, by comparing control and Jmjd3cKO motor neuron lineage cells, we found many downregulated and upregulated genes affected by Jmjd3 loss of function. Single cell RNAseq provided very powerful tool to analyze different regulated genes in each subtype rather than overall motor neuron groups. We used Olig2-Cre and mTmG mouse lines to label Olig2-expressing lineage cells which mainly include motor neurons at E12.5 stage and then employed single cell RNAseq of GFP+ cells to profile transcriptomes in control and Jmjd3cKO mice.为阐明Jmjd3在运动神经元发育中的功能，我们分离了源自表达Olig2的祖细胞（该祖细胞群体包含运动神经元）的绿色荧光蛋白阳性（GFP+）细胞，随后采用单细胞RNA测序（single cell RNAseq）技术开展分析。通过无偏倚且全面的系统性分析，我们不仅鉴定出已知的运动神经元柱（motor columns），还发现了若干新型运动神经元亚型，并明确了各亚型的特异性表达基因。此外，通过对比野生型对照与Jmjd3条件性敲除（Jmjd3cKO）的运动神经元谱系细胞，我们筛选出诸多受Jmjd3功能缺失调控的上调与下调基因。相较于整体运动神经元群体，单细胞RNA测序为解析各亚型的差异调控基因提供了极为高效的研究工具。本研究使用Olig2-Cre与mTmG小鼠品系，在胚胎发育第12.5天（E12.5）标记表达Olig2的谱系细胞（该群体主要包含运动神经元），随后对GFP+细胞进行单细胞RNA测序，以解析野生型对照及Jmjd3cKO小鼠的细胞转录组特征。