Gene expression analysis following neurogenesis of adult enteric glia/progenitor cells

Enteric glia cells (EGCs) are highly plastic and has been shown to give rise to neurons in adult gut upon injury to the enteric nervous system (ENS). To better characterise the dynamic molecular changes associated with EGC activation and its subsequent neurogenic differentation trajectory, we performed bulk RNAseq experiments from FACS sorted lineage-labelled cells (Sox10::CreERT2;R26-tdTomato) from adult small intestines (DIV0) and compared their transcriptomic profiles to cultured FACS sorted tdTomato+ cells from days in vitro (DIV) 4, 11 and 20, which were subjected to a protocol to support neurogenesis in culture. Overall design: Longitudinal muscle-myenteric plexus (LMMP) peels were isolated, washed and dissociated to release adult EGCs for FACS sorting for direct RNA isolation or subjected to a neuronal differentiation protocol before purification by FACS and subsequent RNA isolation. The dynamics of gene expression changes were analysed as a time course, where cultured cells were collected from DIV 4, 11 and 20, compared to samples collected directly from the adult tissues (DIV0).