Differential gene expression and functional analysis implicate novel mechanisms in enteric nervous system precursor migration and neuritogenesis

• DOI: 10.1016/j.ydbio.2006.06.033
• Publication Date: 2006-07-12T13:08:37Z
• Authors: Bhupinder P.S. Vohra; Keiji Tsuji; Mayumi Nagashimada; Toshihiro Uesaka; Daniel Wind; Ming Fu; Jennifer Armon; Hideki Enomoto; Robert O. Heuckeroth
• Keywords: 0301 basic medicine; Cadherin Related Proteins; Gene Expression; Enteric Nervous System; Epithelium; Mice; 03 medical and health sciences; Organ Culture Techniques; Bird; Cell Movement; Morphogenesis; Neurites; Animals; Hirschsprung Disease; Intestinal Mucosa; Protein Precursors; Molecular Biology; Cells, Cultured; Oligonucleotide Array Sequence Analysis; Mice, Knockout; 0303 health sciences; Enteric nervous system development; Proto-Oncogene Proteins c-ret; Cell Biology; Cadherins; 3. Good health; Mice, Inbred C57BL; Chromosome Pairing; SNARE; Neural crest migration; Developmental Biology

Enteric nervous system (ENS) development requires complex interactions between migrating neural-crest-derived cells and the intestinal microenvironment. Although some molecules influencing ENS development are known, many aspects remain poorly understood. To identify additional molecules critical for ENS development, we used DNA microarray, quantitative real-time PCR and in situ hybridization to compare gene expression in E14 and P0 aganglionic or wild type mouse intestine. Eighty-three genes were identified with at least two-fold higher expression in wild type than aganglionic bowel. ENS expression was verified for 39 of 42 selected genes by in situ hybridization. Additionally, nine identified genes had higher levels in aganglionic bowel than in WT animals suggesting that intestinal innervation may influence gene expression in adjacent cells. Strikingly, many synaptic function genes were expressed at E14, a time when the ENS is not needed for survival. To test for developmental roles for these genes, we used pharmacologic inhibitors of Snap25 or vesicle-associated membrane protein (VAMP)/synaptobrevin and found reduced neural-crest-derived cell migration and decreased neurite extension from ENS precursors. These results provide an extensive set of ENS biomarkers, demonstrate a role for SNARE proteins in ENS development and highlight additional candidate genes that could modify Hirschsprung's disease penetrance.