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neurogenetics

, Volume 11, Issue 2, pp 163–174 | Cite as

Progressive retinal atrophy in Schapendoes dogs: mutation of the newly identified CCDC66 gene

  • Gabriele Dekomien
  • Conni Vollrath
  • Elisabeth Petrasch-Parwez
  • Michael H. Boevé
  • Denis A. Akkad
  • Wanda M. Gerding
  • Jörg T. Epplen
ORIGINAL ARTICLE

Abstract

Canine generalized progressive retinal atrophy (gPRA) is characterized by continuous degeneration of photoreceptor cells leading to night blindness and progressive vision loss. Until now, mutations in 11 genes have been described that account for gPRA in dogs, mostly following an autosomal recessive inheritance mode. Here, we describe a gPRA locus comprising the newly identified gene coiled-coil domain containing 66 (CCDC66) on canine chromosome 20, as identified via linkage analysis in the Schapendoes breed. Mutation screening of the CCDC66 gene revealed a 1-bp insertion in exon 6 leading to a stop codon as the underlying cause of disease. The insertion is present in all affected dogs in the homozygous state as well as in all obligatory mutation carriers in the heterozygous state. The CCDC66 gene is evolutionarily conserved in different vertebrate species and exhibits a complex pattern of differential RNA splicing resulting in various isoforms in the retina. Immunohistochemically, CCDC66 protein is detected mainly in the inner segments of photoreceptors in mouse, dog, and man. The affected Schapendoes retina lacks CCDC66 protein. Thus this natural canine model for gPRA yields superior potential to understand functional implications of this newly identified protein including its physiology, and it opens new perspectives for analyzing different aspects of the general pathophysiology of gPRA.

Keywords

CCDC66 gene Insertion mutation Generalized progressive retinal atrophy Retinal protein expression CCDC66 immunohistochemistry 

Notes

Acknowledgements

We thank the owners of the dogs for blood samples, especially H. Mohr and G. de Wit-Bazelmans for their support; the veterinarians of the Dortmunder Ophthalmologenkreis (DOK; Dr. R. Brahm) for the ophthalmologic examinations and enucleation; J. Rutten who carried out parts of the fine mapping and the analysis of candidate genes; and S. Oberland who investigated the C3ORF63 gene. We thank K. Rumpf, A. Schlichting, and H.-W. Habbes for their excellent technical assistance and B. Manderson for proofreading. These studies were supported in part by the Gesellschaft für kynologische Forschung (Bonn, Germany).

Supplementary material

10048_2009_223_MOESM1_ESM.doc (352 kb)
Supplementary Table 1 a: Oligonucleotides used for mutation screening, DNA sequencing, splicing, cloning and qRT-PCR analyses in dog and mouse. *#: Oligonucleotides were also used for splice analysis (DOC 352 kb)
10048_2009_223_MOESM2_ESM.doc (60 kb)
Supplementary Table 2 b: Oligonucleotides used for linkage analysis and fine mapping of the candidate region. F primers were tailed using the unique sequence CAT CGC TGA TTC GCA for HEX or FAM labeling needed for genotyping analyses (DOC 60 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Gabriele Dekomien
    • 1
  • Conni Vollrath
    • 1
  • Elisabeth Petrasch-Parwez
    • 2
  • Michael H. Boevé
    • 3
  • Denis A. Akkad
    • 1
  • Wanda M. Gerding
    • 1
  • Jörg T. Epplen
    • 1
  1. 1.Department of Human GeneticsRuhr-University BochumBochumGermany
  2. 2.Department of Neuroanatomy and Molecular Brain ResearchRuhr-University BochumBochumGermany
  3. 3.Department of Clinical Sciences of Companion Animals, Faculty of Veterinary MedicineUniversity of UtrechtUtrechtThe Netherlands

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