Single-cell RNA Sequencing On CD133+ Cells And Its Differentiation Lineage Isolated From The Fourth Ventricle Of Mice

Neural stem cells(NSCs)is a class of cells with self-renewing capacity,and can differentiate into astrocytes,oligodendrocytes,and neurons.NSCs play an significant role in the development of nervous system and the maintenance of nervous system homeostasis.At present,it is widely demonstrated d that NSCs are mainly distributed in the subventricular zone(SVZ)and the subgranular zone(SGZ).In our previous research,we discovered that the existence of dormant NSCs in the fourth ventricle that can be activated by VEGF cytokine.In brain injury and neurodegenerative diseases,neural stem cells can repair the nervous system by differentiation and migration.Therefore,the study on the expression profile of cells in the fourth ventricle ventricular zone of mice is very important to understand the development of nervous system and the mechanism of neurological diseases.This study used CD133+ neural stem cell lineage tracer mice as experimental material,After isolating the CD133+ NSCs and its daughter cells,we used optimized SMART-seq2 to achieve single-cell c DNA libraries.The sequencing libraries were constructed by Tn5 transposase based tagmentattion method,and the sequencing procedure was accomplished on Illumina sequencing platform.Bioinformatics analysis showed that our mapping rate was up to 91.02%,and 6744 genes per cell on average were detected.We revealed four heterogeneous cell groups in the fourth ventricular ependymal region of mice.Through WGCNA,GO and KEGG pathway analysis,we found significant differences in the metabolic intensity and differentiation stages of these four cell types.Furthermore,We identified cell type ? as a special cell group with features of high metabolic intensity and cell differentiation,and found out signaling pathways associated with cell differentiation including Rap1,Ras,and MAPK signaling pathway.This study provides a high-quality and low-cost scheme for the study of single-cell transcriptome of mouse nerve cells,and brings insight for understanding the heterogeneous cells in the ependymal area of the fourth ventricle.These results are significant to understand the differentiation mechanism of CD133+ neural stem cells,and may provide meaningful ideas for the mechanism research of related neurological diseases.