Diversity of developing peripheral glia revealed by single cell RNA sequencing

We collected dorsal root ganglion (DRG), sciatic nerve (SN) and cochlea tissues from each E14, E18 and P14 plp-GFP+ mice. This allowed for a comparison of plp-GFP+ glia from lumbar (DRG+SN) and cochlea tissues within the same mice. The transcriptional composition of all samples was assessed using single-cell RNA-seq (scRNA-seq). Lumbar tissues (DRG+SN) and cochlea were enzymatically dissociated, GFP+ glia were FACS sorted and profiled by Indrop Single cell v3 method.我们从每只E14、E18及P14期的plp-GFP阳性（plp-GFP+）小鼠中采集了背根神经节（dorsal root ganglion, DRG）、坐骨神经（sciatic nerve, SN）与耳蜗组织。该采样策略可实现同一只小鼠腰段（DRG+SN）与耳蜗来源的plp-GFP+神经胶质细胞的对比研究。所有样本的转录组构成均通过单细胞RNA测序（single-cell RNA-seq, scRNA-seq）进行评估。对腰段组织（DRG+SN）及耳蜗组织进行酶解解离，通过荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）富集GFP+神经胶质细胞，并采用Indrop单细胞v3测序方法完成转录组分析。