title: Characterization of a new, dominant V124E mutation in the mouse alphaA-crystallin-encoding gene.
abstract: OBJECTIVE
  During an ethylnitrosourea (ENU) mutagenesis screening, mice were tested for the occurrence of dominant cataracts. The purpose of the study was morphologic description, mapping of the mutant gene, and characterization of the underlying molecular lesion in a particular mutant, Aey7.
  
  
  METHODS
  Isolated lenses were photographed and histologic sections of the eye were analyzed according to standard procedures. Linkage analysis was performed with a set of microsatellite markers covering all autosomal chromosomes. cDNA was amplified after reverse transcription of lens mRNA. For PCR, cDNA or genomic DNA was used as a template.
  
  
  RESULTS
  Nuclear opacity and posterior suture anomaly were visible at eye opening and progressed to a nuclear and zonular cataract at 2 months of age. The opacity as well as the microphthalmia was more pronounced in the homozygotes than in the heterozygotes. The mutation was mapped to chromosome 17 between the markers D17Mit133 and D17Mit180. This position made the alphaA-crystallin-encoding gene (Cryaa) an excellent candidate gene. Sequence analysis revealed a mutation of a T to an A at position 371 in the Cryaa cDNA. The mutation was confirmed by an additional MnlI restriction site in the genomic DNA of homozygous mutants leading to replacement of Val with Glu at codon 124 affecting the C-terminal region of the alphaA-crystallin.
  
  
  CONCLUSIONS
  The Aey7 mutant represents the first dominant mouse cataract mutation affecting the Cryaa gene. The mutation leads to progressive opacification of the lens. Compared with the beta- and gamma-crystallin-encoding genes, mutations in the alpha-crystallin-encoding genes are rare.
authors.affiliation: National Research Center for Environment and Health (GSF), Institute of Mammalian Genetics, Neuherberg, Germany. graw@gsf.de
authors.forename: J
authors.initials: J
authors.lastname: Graw
authors.forename: J
authors.initials: J
authors.lastname: Löster
authors.forename: D
authors.initials: D
authors.lastname: Soewarto
authors.forename: H
authors.initials: H
authors.lastname: Fuchs
authors.forename: B
authors.initials: B
authors.lastname: Meyer
authors.forename: A
authors.initials: A
authors.lastname: Reis
authors.forename: E
authors.initials: E
authors.lastname: Wolf
authors.forename: R
authors.initials: R
authors.lastname: Balling
authors.forename: M
authors.initials: M
authors.lastname: Hrabé de Angelis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002386:Cataract / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002874:Chromosome Mapping; D002875:Chromosomes / Q000235:genetics; D003459:Crystallins / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D005799:Genes, Dominant; D008040:Genetic Linkage; D007908:Lens, Crystalline / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D003459:Crystallins; D017931:DNA Primers