TIMP-1 expression is increased in X-linked progressive retinal atrophy despite its exclusion as a candidate gene.

ABSTRACT

X-linked progressive retinal atrophy (XLPRA) is the only known natural animal model for X-linked retinitis pigmentosa (XLRP), a blinding disorder in man. The tissue inhibitor metalloproteinase 1 gene (TIMP-1), present in close proximity to one of the two XLRP loci, was tested as a candidate for XLPRA, by first characterizing the cDNA and gene from a normal dog. The cloned canine TIMP-1 cDNA is predicted to encode a protein of 207 amino acids with 66-83% identity in the deduced aa sequence with homologous mammalian genes. No sequence difference in the coding sequence of TIMP-1 was observed between normal and XLPRA-affected dogs. TIMP-1 was found to be expressed in all of the canine tissues examined by reverse transcription and polymerase chain reaction. The canine TIMP-1 spans 3.5kb and is interrupted by five introns with sizes comparable to those observed in the human and mouse homologues of the gene. The proximal promoter region of canine TIMP-1 contains sequence motifs shown to have regulatory significance in transcription of human TIMP-1. Linkage analysis between XLPRA and TIMP-1 using a newly identified intragenic polymorphism identified recombinants, which conclusively excluded the gene as a candidate for the disease. TIMP-1 is overexpressed several months before retinal degeneration is histologically evident in XLPRA dogs, implying that alterations in interphotoreceptor matrix composition precede retinal degeneration by a significant time period.