title: Characterization of the genetic basis for autosomal recessive hereditary nephropathy in the English Springer Spaniel.
abstract: BACKGROUND
  Autosomal recessive hereditary nephropathy (ARHN) was diagnosed in 2 English Springer Spaniels (ESS), a breed not previously reported to be affected by hereditary nephropathy (HN).
  
  
  OBJECTIVE
  To identify and characterize the genetic cause of ARHN in ESS.
  
  
  METHODS
  Sixty-three ESS (2 with ARHN, 2 obligate carriers, and 59 others), 2 mixed-breed dogs with X-linked HN, and 2 English Cocker Spaniels (ECS) with ARHN were included.
  
  
  METHODS
  ARHN was diagnosed based on transmission electron microscopy and immunostaining of kidney. DNA from affected dogs was screened for the mutation known to cause ARHN in ECS. Quantities of COL4A3, COL4A4, and COL4A5 mRNA transcripts in renal cortex were determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for ARHN-affected dogs and 7 other dogs. The coding regions of COL4A3 and COL4A4 were sequenced for the 2 ARHN-affected ESS and an unaffected dog. Exon 30 of COL4A4 was sequenced for all 63 ESS.
  
  
  RESULTS
  qRT-PCR indicated a significant reduction in transcript levels of both COL4A3 and COL4A4 mRNA in the kidney of ARHN-affected ESS. Sequencing identified a single nucleotide substitution in COL4A4 at base 2806 resulting in a premature stop codon. Thirteen of 25 related dogs were identified as carriers.
  
  
  CONCLUSIONS
  A mutation highly likely to cause ARHN in ESS has been identified.
authors.affiliation: Department of Genetics and Biochemistry, Clemson University, Clemson, SC 29634-0318, USA.
authors.forename: K L
authors.initials: KL
authors.lastname: Nowend
authors.forename: A N
authors.initials: AN
authors.lastname: Starr-Moss
authors.forename: G E
authors.initials: GE
authors.lastname: Lees
authors.forename: B R
authors.initials: BR
authors.lastname: Berridge
authors.forename: F J
authors.initials: FJ
authors.lastname: Clubb
authors.forename: C E
authors.initials: CE
authors.lastname: Kashtan
authors.forename: M B
authors.initials: MB
authors.lastname: Nabity
authors.forename: K E
authors.initials: KE
authors.lastname: Murphy
mesh_terms: D000818:Animals; D001483:Base Sequence; D024141:Collagen Type IV / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004283:Dog Diseases / Q000235:genetics* / Q000473:pathology; D004285:Dogs; D005260:Female; D014644:Genetic Variation; D007678:Kidney Glomerulus / Q000473:pathology / Q000648:ultrastructure; D008297:Male; D046529:Microscopy, Electron, Transmission / Q000662:veterinary; D008969:Molecular Sequence Data; D009394:Nephritis, Hereditary / Q000235:genetics / Q000473:pathology / Q000662:veterinary*; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA
chemical_list: D024141:Collagen Type IV; D018076:DNA, Complementary