Cloning and characterization of opticin cDNA: evaluation as a candidate for canine oculo-skeletal dysplasia

Abstract

Opticin, a novel member of the leucine-rich repeat (LRR) family, has been reported to bind to collagen fibrils. Many members of the LRR family of extracellular matrix proteins have been reported to bind to fibrillar collagen and regulate the diameter of collagen fibrils and lateral fusion between fibrils. Collagen fibrils are important for the maintenance of the vitreous body in the eye and growth plate cartilage of joints. Oculo-skeletal dysplasia (OSD) is a heterogeneous group of heritable genetic disorders affecting humans and a few breeds of dogs. Labrador retrievers and Samoyeds affected with non-allelic forms of OSD exhibit vitreous dysplasia and dwarfism, and could serve as an animal model for the disorder. To test the opticin gene as a candidate for OSD, canine opticin cDNA has been cloned and characterized. The predicted 327 amino acid sequence is 77% homologous to human opticin, and maintains characteristic structural domains including seven LRR domains, two cysteine clusters and potential O-linked glycosylation sites. It shows highest protein sequence identity to epiphycan (37%) and osteoglycin (31%) and belongs to the Class III family of LRR extracellular matrix proteins. In addition to ocular tissues and cartilage, opticin mRNA and protein have been identified in ligament, skin, muscle, and testes. No alteration of opticin expression at the protein level was observed in OSD affected dogs relative to normal controls. Based on linkage analysis using a newly identified intragenic single nucleotide polymorphism opticin has been excluded from having any causal association with the OSD loci in both Samoyeds and Labrador retrievers.