Features of glial cell clusters after peripheral nerve injury based on single-cell RNA sequencing
10.3760/cma.j.cn115354-20240712-00408
- VernacularTitle:基于scRNA-seq技术分析周围神经损伤后胶质细胞亚群特点的实验研究
- Author:
Jinsheng HUANG
1
;
Laijin LU
;
Nan ZHOU
Author Information
1. 郑州大学第一附属医院骨科，河南 450000
- Keywords:
Peripheral nerve injury;
Single-cell RNA sequencing;
Glial cell;
Cell atlas
- From:
Chinese Journal of Neuromedicine
2024;23(9):865-873
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the species and number of glial cell clusters, main signaling pathway and progression of glial cell clusters after peripheral nerve injury (PNI) in rats using single-cell RNA sequencing (scRNA-seq).Methods:Twenty-seven SD rats were randomly divided into sham-operated group, group of 3 d after PNI and group of 7 d after PNI ( n=9). Thrice squeezing the right sciatic nerves in the group of 3 d after PNI and group of 7 d after PNI and no damage in the sham-operated group were performed. Species and number of glial cell clusters in the right sciatic nerve samples were detected by scRNA-seq. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the signal pathway enriched by differentially expressed genes (DEGs) of glial cells in the right sciatic nerve samples. Pseudo time analysis was used to simulate the progression of glial cell clusters in the right sciatic nerve samples. Results:(1) ScRNA-seq revealed a total of 1 609 glial cells (mainly cluster 6 [ n=1 388]) in the sciatic nerve samples of sham-operated group; 6 176 glial cells were observed in the sciatic nerve samples of group of 3 d after PNI, mainly cluster 2 ( n=3 124) and cluster 3 ( n=959); 8 975 glial cells were observed in the sciatic nerve samples of group of 7 d after PNI, mainly cluster 1 ( n=3 071), cluster 4 ( n=1 696), and cluster 5 ( n=1 389). (2) GO and KEGG analysis showed that compared with those in the sham-operated group, biological processes such as protein translation, cadherin binding and ribosome composition were up-regulated in glial cells in the group of 3 d after PNI and group of 7 d after PNI. Compared with those in the group of 3 d after PNI, glial cells enriched in biological processes such as axonal regeneration, myelination and focal adhesion, and in upregulated mitogen activated protein kinase (MAPK) signaling pathway and phosphoinositide 3-kinase (PI3K)-protein kinase B (PKB) signaling pathway in the group of 7 d after PNI. (3) Pseudo time analysis showed that glial cells in the sciatic nerve samples were mainly cluster 6 (marker genes: Atp1a2 and Sparcl1) in the sham-operated group, progressed into cluster 2 (marker genes: Mapt and Slc7a11) and cluster 3 (marker genes: Esco2 and Neil3) mainly in the group of 3 d after PNI, and progressed into cluster 1 (marker genes: Bcas1 and Prx), cluster 4 (marker genes: Ccn2 and Gap43), and cluster 5 (marker genes: Cd24 and Atxn1) mainly in the group of 7 d after PNI. Conclusion:In rats after PNI, glial cells can up-regulate MAPK signaling pathway and PI3K-PKB signaling pathway; with prolonged injury time, glial cells can progress into clusters with marker genes Bcas1, Prx, Cd24 and Atxn1 mainly.
