Danforth's short tail (Sd) is a semidominant mutation on mouse chromosome 2, characterized by spinal defects, urogenital defects, and anorectal malformations. However, the gene responsible for the Sd phenotype was unknown. In this study, we identified the molecular basis of the Sd mutation. By positional cloning, we identified the insertion of an early transposon in the Sd candidate locus approximately 12-kb upstream of Ptf1a. We found that insertion of the transposon caused overexpression of three neighboring genes, Gm13344, Gm13336, and Ptf1a, in Sd mutant embryos and that the Sd phenotype was not caused by disruption of an as-yet-unknown gene in the candidate locus. Using multiple knockout and knock-in mouse models, we demonstrated that misexpression of Ptf1a, but not of Gm13344 or Gm13336, in the notochord, hindgut, cloaca, and mesonephros was sufficient to replicate the Sd phenotype. The ectopic expression of Ptf1a in the caudal embryo resulted in attenuated expression of Cdx2 and its downstream target genes T, Wnt3a, and Cyp26a1; we conclude that this is the molecular basis of the Sd phenotype. Analysis of Sd mutant mice will provide insight into the development of the spinal column, anus, and kidney.
Caudal regression syndrome (CRS) is a congenital heterogeneous constellation of caudal anomalies that includes varying degrees of agenesis of the spinal column, anorectal malformations, and genitourinary anomalies. Its pathogenesis is unclear. However, it could be the result of excessive physiologic regression of the embryonic caudal region based on analyses of the various mouse mutants carrying caudal agenesis. Among the mouse mutants, the Danforth's short tail (Sd) mouse is considered a best model for human CRS. Sd is a semidominant mutation, characterized by spinal defects, urogenital defects, and anorectal malformations, thus showing phenotypic similarity to human CRS. Although Sd is known to map to mouse chromosome 2, little is known about the molecular nature of the mutation. Here, we demonstrate an insertion of one type of retrotransposon near the Ptf1a gene. This resulted in ectopic expression of Ptf1a gene in the caudal region of the embryo and downregulation of Cdx2 and its downstream targets, leading to characteristic phenotypes in Sd mouse. Thus, Sd mutant mice will provide insight into the development of the spinal column, anus, and kidney.