Expression profiling the developing mammalian enteric nervous system identifies marker and candidate Hirschsprung disease genes.
The enteric nervous system (ENS) is composed of neurons and glial cells, organized as interconnected ganglia within the gut wall, which controls peristalsis of the gut wall and secretions from its glands. The Ret receptor tyrosine kinase is expressed throughout enteric neurogenesis and is required for normal ENS development; humans with mutations in the RET locus have Hirschsprung disease (HSCR, an absence of ganglia in the colon), and mice lacking Ret have total intestinal aganglionosis. The Ret mutant mouse provides a tool for identifying genes implicated in development of the ENS. By using RNA from WT and Ret mutant (aganglionic) gut tissue and DNA microarrays, we have conducted a differential screen for ENS-expressed genes and have identified hundreds of candidate ENS-expressed genes. Forty-seven genes were selected for further analysis, representing diverse functional classes. We show that all of the analyzed genes are expressed in the ENS and that the screen was sensitive enough to identify genes marking only subpopulations of ENS cells. Our screen, therefore, was reliable and sensitive and has identified many previously undescribed genes for studying ENS development. Moreover, two of the genes identified in our screen Arhgef3 and Ctnnal1, have human homologues that map to previously identified HSCR susceptibility loci, thus representing excellent candidates for HSCR genes. This comprehensive profile of ENS gene expression refines our understanding of ENS development and serves as a resource for future developmental, biochemical, and human genetic studies.
Heanue
Tiffany A
TA
Division of Molecular Neurobiology, National Institute for Medical Research, Medical Research Council, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom.
Pachnis
Vassilis
V
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D041321:Chromosomes, Human, X; D017615:Enteric Nervous System / Q000033:anatomy & histology / Q000254:growth & development* / Q000502:physiology*; D041981:Gastrointestinal Tract / Q000033:anatomy & histology / Q000254:growth & development / Q000502:physiology; D020869:Gene Expression Profiling; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism; D006627:Hirschsprung Disease / Q000235:genetics*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics / Q000378:metabolism; D064067:Rho Guanine Nucleotide Exchange Factors; D051178:alpha Catenin / Q000235:genetics / Q000378:metabolism
C576601:ARHGEF3 protein, human; D015415:Biomarkers; C495274:CTNNAL1 protein, human; D005819:Genetic Markers; D020662:Guanine Nucleotide Exchange Factors; D064067:Rho Guanine Nucleotide Exchange Factors; D051178:alpha Catenin; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human