Dynamic Changes in the Proximal Gut Neural Crest Stem Cell Population Are Associated with Successful Development of the Distal Enteric Nervous System in Rats

Abstract

Loss of signaling through the endothelin-B receptor (ET(B)) leads to failure of vagal neural crest (NC) cell colonization of the developing gut and causes congenital distal intestinal aganglionosis [Hirschsprung disease (HSCR)] in humans and other mammals. Several studies suggest that cell-cell interactions and the number of NC cells behind the wavefront may play an important role in successful gut colonization. We compared the number and progression of enteric nervous system stem cells in the wild-type (WT) and HSCR rat gut using whole-mount immunohistochemistry for p75, culture and isolation of NC stem cells (NCSCs) by flow cytometry. Isolation and culture demonstrates that NCSCs enter the WT cecum between embryonic day (E) 13.5 and E14.5, and the number of NCSC in the colon significantly increases after E15.5. These findings are consistent with the caudal progression of the NC-cell wavefront by whole-mount staining. During the period of WT colonic colonization of the proximal colon, we found significant differences in the small bowel NCSC pool between WT and HSCR rats. Whereas the proximal gut NCSC pool in WT rats is increasing behind the colonization wavefront, no such change occurs in the proximal NCSC pool in HSCR rats. Dynamic changes in the NCSC pool occur behind the NC colonization wavefront in the gut of WT rats. The absence of these changes in the HSCR rat may contribute to distal aganglionosis.