A goal of our research is to characterize genetic organization in the domestic cat and to develop genomic resources facilitating and establishing, Felis catus, as a useful animal model contributing to our understanding of human hereditary disease analogues, neoplasia, genetic factors associated with host response to infectious disease and mammalian genome evolution. In order to map and characterize genes associated with inherited pathologies in the domestic cat which mirror inherited human conditions we have focused on increasing microsatellite density in the genetic map of the cat to increase resolution for mapping of genes associated with inherited and infectious disease. We are currently focused on completion of a third generation full genome genetic linkage map of microsatellites in the cat, with marker resolution of approximately 3 cM. This map will provide us with a sufficient resolution to perform linkage-mapping exercises in several cat pedigrees of interest segregating for disease phenotype and phenotypes of other biological interest. Approximately 700 microsatellites, described above, are being genotyped in a large multi-generation domestic cat pedigree maintained by the Nestle-Purina Pet Care Company (n=483 informative meioses). Based on previous mapping of a majority of these loci in the cat radiation-hybrid panel, we know that the loci are well distributed across the cat genome. Currently approximately 700 loci have been amplified and genotyped in the multigeneration pedigree. Mendelian inheritance checking is currently in progress and necessary before markers can be ordered with respect to one another. Mapping and Characterization of Genes Associated with Inherited Disease Pathology in Cat Pedigrees with Homology to Human Hereditary Disease: 1. Autosomal recessive retinitis pigmentosa (arRP) is a genetically and clinically heterogeneous and progressive degenerative disorder of the retina, leading usually to severe visual handicap in adulthood. Our collaborator, Dr. Kristina Narfstrom at the Missouri College of Veterinary Medicine, maintains a colony of Abyssinian cats with progressive retinal atrophy (rdAc), a slowly progressive degeneration process of the rod and cone systems with similarities to classical human retinitis pigmentosa (RP). The Abyssinian cat has the potential of becoming a new and important animal model in the study of hereditary visual cell disease processes. In order to identify and characterize the gene which gives rise to feline rdAc, our approach is to map the gene using classical linkage mapping with a full genome scan of Dr. Narfstroms's pedigree. As initial screening of the pedigree with 254 microsatellite loci exhibited that the colony was highly inbred, Dr. Narfstrom outbred members of the pedigree with controlled matings to increase the average marker heterozygosity. To date, a total of 52 backcross individuals have been generated from 18 outbred F1, and more individuals are being generated. Simulation results (SIMLINK) indicate that the likelihood of detecting linkage to the disease locus in a pedigree structure/size similar to Dr. Narfstrom's with 45 or 60 back-crossed individuals is 0.88 and 0.91, respectively, at q = 0.1 (lod score = 5.5, 7.1, respectively) (average heterozygosity of microsatellites = 0.75). 2. Spinal Muscular Atrophy, Degeneration of lower motor neurons in the spinal cord that causes neurogenic muscle atrophy, the so-called spinal muscular atrophies (SMAs), are a large category of inherited disorders, but the molecular basis or linkage of a genetic locus has been established in only a few. The SMAs are categorized generally by pattern of inheritance and by distribution of muscle atrophy. In collaboration with Dr. John Fyfe (Michigan State University), who maintains a pedigree of Maine Coon cats segregating for SMA Type III, we are investigating the molecular basis of this new and unique model of inherited motor neuron degeneration in cats. Our prior linkage analysis has indicated that disease phenotypes are unlinked to the feline spinal muscular neuron (SMN) locus. A full genome scan of microsatellites is currently being performed in his affected pedigree. Once linkage is established to a cat chromosome, comparative gene mapping will be utilized to identify candidate genes for the pathology. 3. Completion of an single tandem repeat (STR) Forensic Typing System for Genetic Individualization of Domestic Cat Specimens. Under support from the National Institute of Justice (NIJ), we are completing the development of a microsatellite forensic typing system and genetic database of cat breeds for genetic individualization of cat specimens. A panel of 11 tetra-nucleotide repeat microsatellite loci has been isolated, mapped in the cat radiation hybrid (RH) map and examined for Mendelian inheritance and mutation rate. A robust multiplex amplification protocol has been developed, validated and genotyped in a panel of 1043 cats representing 38 recognized breeds in the United States. The multiplex demonstrates high discriminating power in cat breeds, exhibiting average locus heterozygosity across the 38 breeds of 0.71. A real-time polymerase chain reaction (PCR) based method has been developed for quantifying domestic cat genomic DNA, which is sensitive to 10 femtograms.
|Troyer, Jennifer L; Brown, Meredith A (2011) Feline models of viral pathogenesis: opportunity knocks. Vet J 188:252-3|
|Coomber, Nikia; David, Victor A; O'Brien, Stephen J et al. (2007) Validation of a short tandem repeat multiplex typing system for genetic individualization of domestic cat samples. Croat Med J 48:547-55|
|Ishida, Yasuko; David, Victor A; Eizirik, Eduardo et al. (2006) A homozygous single-base deletion in MLPH causes the dilute coat color phenotype in the domestic cat. Genomics 88:698-705|
|Fyfe, John C; Menotti-Raymond, Marilyn; David, Victor A et al. (2006) An approximately 140-kb deletion associated with feline spinal muscular atrophy implies an essential LIX1 function for motor neuron survival. Genome Res 16:1084-90|
|He, Qianchuan; Lowrie, Charles; Shelton, G Diane et al. (2005) Inherited motor neuron disease in domestic cats: a model of spinal muscular atrophy. Pediatr Res 57:324-30|
|Schmidt-Kuntzel, A; Eizirik, E; O'Brien, S J et al. (2005) Tyrosinase and tyrosinase related protein 1 alleles specify domestic cat coat color phenotypes of the albino and brown loci. J Hered 96:289-301|