title: Genetic cause of autosomal recessive hereditary nephropathy in the English Cocker Spaniel.
abstract: BACKGROUND
  Autosomal recessive hereditary nephropathy (ARHN) in the English Cocker Spaniel is caused by a type IV collagen defect, but the underlying mutation is unknown.
  
  
  METHODS
  One hundred thirty-four English Cocker Spaniels (12 with ARHN, 8 obligate carriers, and 114 others), 3 mixed breed dogs with X-linked hereditary nephropathy (XLHN), and 7 other dogs without hereditary nephropathy were included.
  
  
  METHODS
  Diagnosis of ARHN was based on transmission electron microscopy and immunostaining of kidney. Quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR) was used to compare COL4A3, COL4A4, and COL4A5 mRNA concentrations in the renal cortex from ARHN-affected English Cocker Spaniels, XLHN-affected dogs, and dogs without hereditary nephropathy. The entire coding region of COL4A4 was sequenced in 2 ARHN-affected dogs, 2 obligate carriers, 2 English Cocker Spaniels of unknown status, and 2 healthy mixed breed dogs. The exon containing the mutation was sequenced for all 134 English Cocker Spaniels.
  
  
  RESULTS
  Quantitative real time RT-PCR implicated COL4A4 as the gene harboring the mutation, and sequencing identified a single nucleotide substitution at base 115 as the cause of ARHN in English Cocker Spaniels. This mutation, which causes a premature stop codon in exon 3 of COL4A4, was segregated with clinical status in all affected dogs and obligate carriers. The mutation also was identified in 39 of 114 other English Cocker Spaniels with previously unknown status.
  
  
  CONCLUSIONS
  The cause of this disease has been identified, and use of a test for the mutation will permit eradication of ARHN in the English Cocker Spaniel.
authors.affiliation: Department of Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843-4467, USA.
authors.forename: Ashley G
authors.initials: AG
authors.lastname: Davidson
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Bell
authors.forename: George E
authors.initials: GE
authors.lastname: Lees
authors.forename: Clifford E
authors.initials: CE
authors.lastname: Kashtan
authors.forename: George S
authors.initials: GS
authors.lastname: Davidson
authors.forename: Keith E
authors.initials: KE
authors.lastname: Murphy
mesh_terms: D000818:Animals; D001483:Base Sequence; D003937:Diagnosis, Differential; D004283:Dog Diseases / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D004285:Dogs; D005091:Exons; D005260:Female; D040181:Genetic Diseases, X-Linked / Q000662:veterinary*; D007150:Immunohistochemistry / Q000662:veterinary*; D007668:Kidney / Q000473:pathology / Q000648:ultrastructure*; D007674:Kidney Diseases / Q000175:diagnosis / Q000235:genetics / Q000473:pathology / Q000662:veterinary*; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary