Impact of Enteric Neuronal Loss on Intestinal Cell Composition

This study describes the intestinal composition in wildtype and ret mutant zebrafish intestines using single cell RNA sequencing (scRNA-seq). Overall design: In this study, we investigated the impact of ENS loss on intestinal composition using a rethu2846/hu2846 mutant tg(phox2bb:GFP) zebrafish. At 4 dpf, rethu2846/hu2846 mutant tg(phox2bb:GFP) zebrafish showing a total colonic HSCR phenotype were selected. Zebrafish intestines were manually isolated at 5dpf and dissociated into a single cell suspension. Live, single cells were sorted by Fluorescence-activated cell sorting (FACS) according to the presence or absence of DAPI signal. Sorted cells were processed and analyzed using 10X scRNAseq. To provide a comprehensive analysis, the ret mutant data was integrated with previously published wildtype zebrafish intestines (GSE225510)