title: Histochemical study of Dom mouse: A model for Waardenburg-Hirschsprung's phenotype.
abstract: OBJECTIVE
  The spontaneous mouse mutant Dominant megacolon (Dom) represents the model of the Waardenburg-Hirschsprung's disease, a syndromic pathology, characterized by the association of pigmentation defects (PD), deafness, and Hirschsprung's disease (HD). The defect in Dom mouse is caused by a spontaneous mutation of the gene encoding the Sry-related transcription factor Sox10. This mutation affects several aspects of neural crest development leading to combined enteric innervation and pigmentation defects, both in mouse and human. The purpose of this report is to define, by enzymo-histochemical techniques routinely used for the diagnosis of human Hirschsprung's disease (AChE, LDH, NADPH-diaphorase), the innervative patterns of the affected gut.
  
  
  METHODS
  Fifty-four siblings of Heterozygous Dom/+ mice underwent autopsy and were genotyped by direct sequencing of polymerase chain reaction (PCR) products for Sox10 mutations. The enteric nervous system of all the mice was studied by histochemical techniques indicated above.
  
  
  RESULTS
  Genotyping showed that 43 mice were Dom/+ and 11 were Wild type +/+. Wild-type +/+ mice were used as control. The correspondence between genotype and at least 1 phenotypic aspect (PD or dysganglionosis) was present in 93% of cases (41 of 43). Among the Dom/+ mice, dysganglionosis was present in 79% of cases and PD in 90% of cases. Moreover, among Dom/+ mice, excluding those whose mantle was not evaluated as dead just after birth, PD and dysganglionosis (complete phenotype) were present in 68% of cases.
  
  
  CONCLUSIONS
  The histochemical methods that we used proved to be useful for identification of different aganglionic (AG), hypoganglionic (HG), and normoganglionic segments of Dom/+ mouse gut studied in longitudinal sections. Unlike humans, control mice (Wild type +/+) presented a rich component of AChE nerve fibers, whereas Dom/+ mice with dysganglionosis presented a decrease in AChE-positive nerve fibers. These data confirm the variable phenotypic penetrance in heterozygous mice. Because dysganglionosis in this animal model (Dom/+) was evident in 79% of cases (AG or HG), we concluded that Dom mice could represent important models for further experimental studies.
authors.affiliation: Department of Pediatric Surgery, G. Gaslini Children's Hospital, University of Genova, Genoa, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Brizzolara
authors.forename: Michele
authors.initials: M
authors.lastname: Torre
authors.forename: Anna
authors.initials: A
authors.lastname: Favre
authors.forename: Alessio
authors.initials: A
authors.lastname: Pini Prato
authors.forename: Renata
authors.initials: R
authors.lastname: Bocciardi
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Martucciello
mesh_terms: D000110:Acetylcholinesterase / Q000032:analysis; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D033481:Chromosomes, Mammalian; D003106:Colon / Q000473:pathology; D004195:Disease Models, Animal; D017615:Enteric Nervous System / Q000473:pathology*; D005260:Female; D005838:Genotype; D006627:Hirschsprung Disease / Q000473:pathology*; D018398:Homeodomain Proteins; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D016254:Mutagenesis, Insertional / Q000235:genetics; D009252:NADPH Dehydrogenase / Q000032:analysis; D010641:Phenotype; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics; D014849:Waardenburg Syndrome / Q000473:pathology*
chemical_list: D015415:Biomarkers; D018398:Homeodomain Proteins; C497420:Six2 protein, mouse; D014157:Transcription Factors; D007770:L-Lactate Dehydrogenase; D009252:NADPH Dehydrogenase; D000110:Acetylcholinesterase