Differential gene expression and functional analysis implicate novel mechanisms in enteric nervous system precursor migration and neuritogenesis.
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.
Vohra
Bhupinder P S
BP
Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA.
Tsuji
Keiji
K
Nagashimada
Mayumi
M
Uesaka
Toshihiro
T
Wind
Daniel
D
Fu
Ming
M
Armon
Jennifer
J
Enomoto
Hideki
H
Heuckeroth
Robert O
RO
D000818:Animals; D000070585:Cadherin Related Proteins; D015820:Cadherins / Q000378:metabolism / Q000502:physiology; D002465:Cell Movement; D002478:Cells, Cultured; D023902:Chromosome Pairing; D017615:Enteric Nervous System / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D006627:Hirschsprung Disease / Q000139:chemically induced / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009024:Morphogenesis; D016501:Neurites / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D009924:Organ Culture Techniques; D011498:Protein Precursors / Q000378:metabolism / Q000502:physiology; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics; D050600:SNARE Proteins / Q000502:physiology; D013234:Stem Cells / Q000502:physiology*; D051178:alpha Catenin / Q000378:metabolism / Q000502:physiology
C418976:CDHR15 protein, mouse; D000070585:Cadherin Related Proteins; D015820:Cadherins; D011498:Protein Precursors; D050600:SNARE Proteins; D051178:alpha Catenin; D051096:Proto-Oncogene Proteins c-ret; C494999:Ret protein, mouse