Defective cell adhesion and cell migration in Sox10 mutant neural crest

Abstract

The enteric neural crest cells (ENCCs) are derived from migratory vagal neural crest cells (NCCs). Transcription factor Sox10 is crucial for ENCC multipotency maintenance. Sox10 mutant ENCCs fail to colonize the entire gut due to premature differentiation and migration defects. A Sox10NGFP mouse where EGFP is fused to the N-terminal domain of Sox10 was generated to visualize the migration behavior of mutant ENCCs. Sox10NGFP/+ mutant ENCCs form aggregates on different extracellular matrices in gut explant culture. We hypothesized that cell adhesion properties were affected in Sox10 mutant neural crest. To gain insight into the cell adhesion and molecular pathways underlying cell migration processes in ENCCs, we performed transcriptome and gene expression analysis as well as immunofluorescence analysis to study cellular phenotypes. E9.5 pre-ENCCs and E12.5 ENCCs from Wnt1-Cre:Z/EG and Sox10NGFP mutant were sorted out and performed RNA sequencing. Bioinformatics analysis indicated that among the differentially expressed genes, the expression of biological adhesion and locomotion genes were significantly affected during ENCC development. Selected differentially expressed genes were further verified by qRT-PCR. The immunostaining results of cultured explants showed that the distribution of cadherins, vinculin and FAK were affected in Sox10 mutant NCCs and ENCCs. Further cellular behavior such as real-time actin filament dynamics and intensity will be assessed by live cell imaging using Wnt1-Cre:Z/EG:Lifeact-mRFP and Sox10NGFP:Lifeact-mRFP mice, which had the ENCCs labelled by EGFP and F-actin . Our results suggest that Sox10 mutation alters the expression of cell adhesion molecules and ECM reporters, thereby may affect ENCCs cellular behavior and migration.