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Histochemical study of Dom mouse: A model for Waardenburg-Hirschsprung's phenotype
Antonella Brizzolara1, Michele Torre, Anna Favre
1Department of Pediatric Surgery, G. Gaslini Children's Hospital, University of Genova, Genoa, Italy.
Background/Purpose:
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.
Insights
The Dominant megacolon (Dom) mouse model, caused by a Sox10 mutation, exhibits Hirschsprung
Area of Science:
- Developmental biology
- Genetics
- Neuroscience
Background:
- The Dominant megacolon (Dom) mouse is a model for Waardenburg-Hirschsprung's disease, a condition involving pigmentation defects, deafness, and Hirschsprung's disease.
- The Dom mutation affects the Sox10 gene, crucial for neural crest development, leading to enteric and pigmentation abnormalities in both mice and humans.
Purpose of the Study:
- To characterize the innervative patterns of the affected gut in Dom mice using histochemical techniques.
- To evaluate the utility of these techniques for diagnosing Hirschsprung's disease in this animal model.
Main Methods:
- Genotyping of 54 mouse siblings to identify Dom/+ and wild-type +/+ genotypes.
- Histochemical analysis of the enteric nervous system using acetylcholinesterase (AChE), lactate dehydrogenase (LDH), and NADPH-diaphorase.
Main Results:
- Genotyping confirmed 43 Dom/+ and 11 wild-type mice.
- Phenotypic correlation (pigmentation defects or dysganglionosis) was observed in 93% of Dom/+ mice.
- Dysganglionosis occurred in 79% and pigmentation defects in 90% of Dom/+ mice, with a complete phenotype in 68%.
Conclusions:
- Histochemical methods effectively identified aganglionic, hypoganglionic, and normoganglionic gut segments in Dom/+ mice.
- Dom/+ mice showed reduced AChE nerve fibers compared to controls, confirming variable phenotypic penetrance.
- Dom mice are valuable models for studying Hirschsprung's disease due to frequent dysganglionosis.
