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J. Kuchtey, L. Olson, J. L. Haines, P. J. Dexheimer, S. Levy, E. O. MacKay, K. N. Gelatt, R. W. Kuchtey; Identifying the Gene Causing Hereditary Primary Open Angle Glaucoma in Beagles. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2150.
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© ARVO (1962-2015); The Authors (2016-present)
To identify the disease gene causing hereditary primary open angle glaucoma (POAG) in Beagle dogs.
A colony of Beagles with naturally occurring autosomal recessive glaucoma was founded decades ago and established as an animal model for human POAG. DNA samples were obtained from affected, carrier and normal dogs from the POAG Beagle colony, as well as from normal unrelated Beagles. Affection status of the dogs from the POAG colony was determined by complete eye exams including gonioscopy, measurements of intraocular pressure (IOP) and optic nerve evaluation. Normal Beagles unrelated to the colony were determined to be not affected by glaucoma by clinical exam. Approximately 50,000 single nucleotide polymorphisms (SNPs) covering all canine autosomes were genotyped using SNP microarrays. Selected SNP genotypes were confirmed by PCR/mass-spectrometry analysis. Genome wide SNP genotype data for dogs from the POAG Beagle colony were analyzed by zygosity analysis (identification of SNPs that are both homozygous for all affected and heterozygous for all carriers) and two-point linkage analysis. Genome wide two-point linkage analysis was followed up with multipoint linkage analysis for regions of interest. Microarray-based sequence capture was used to isolate DNA for analysis by high-throughput sequencing.
Zygosity analysis of genome wide SNP data from the POAG Beagle colony identified a single 4 Mb locus likely to contain the disease allele. Two-point linkage analysis of the genome wide data set also identified the same 4 Mb locus and two additional loci. Multipoint linkage analysis and/or haplotype analysis ruled out the two additional loci. DNA libraries representing all non-repetitive DNA within the 4 Mb region were isolated from normal, carrier and affected Beagles for high-throughput DNA sequencing.
A genetic variant responsible for POAG in the Beagles is likely located within the 4 Mb region identified by zygosity and linkage analysis. High-throughput DNA sequencing of the 4 Mb region which has been captured by microarray is being carried out to identify the disease-causing genetic variant.
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