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The  English spotting  coat color locus in rabbits, also known as  Dominant white spotting  locus, is determined by an incompletely dominant allele ( En ). Rabbits homozygous for the recessive wild-type allele ( en / en ) are self-colored, heterozygous  En/en  rabbits are normally spotted, and homozygous  En/En  animals are almost completely white. Compared to vital  en / en  and  En/en  rabbits,  En/En  animals are subvital because of a dilated (“mega”) cecum and ascending colon. In this study, we investigated the role of the  KIT  gene as a candidate for the  English spotting  locus in Checkered Giant rabbits and characterized the abnormalities affecting enteric neurons and c-kit positive interstitial cells of Cajal (ICC) in the megacolon of  En/En  rabbits. Twenty-one litters were obtained by crossing three Checkered Giant bucks ( En/en ) with nine Checkered Giant ( En/en ) and two  en/en  does, producing a total of 138 F1 and backcrossed rabbits. Resequencing all coding exons and portions of non-coding regions of the  KIT  gene in 28 rabbits of different breeds identified 98 polymorphisms. A single nucleotide polymorphism genotyped in all F1 families showed complete cosegregation with the  English spotting  coat color phenotype ( θ  = 0.00 LOD  = 75.56).  KIT  gene expression in cecum and colon specimens of  En/En  (pathological) rabbits was 5–10% of that of  en/en  (control) rabbits.  En/En  rabbits showed reduced and altered c-kit immunolabelled ICC compared to  en/en  controls. Morphometric data on whole mounts of the ascending colon showed a significant decrease of HuC/D ( P <0.05) and substance P ( P <0.01) immunoreactive neurons in  En/En  vs.  en/en . Electron microscopy analysis showed neuronal and ICC abnormalities in  En/En  tissues. The  En/En  rabbit model shows neuro-ICC changes reminiscent of the human non-aganglionic megacolon. This rabbit model may provide a better understanding of the molecular abnormalities underlying conditions associated with non-aganglionic megacolon.

The KIT Gene Is Associated with the English Spotting Coat Color Locus and Congenital Megacolon in Checkered Giant Rabbits (Oryctolagus cuniculus)