Purpose: To determine whether early retinal degeneration (erd) and progressive rod cone degeneration (prcd), two canine hereditary retinal degenerations, are caused by allelic mutations; to determine the cDNA sequence of the canine RDS/peripherin homolog (CFRDSP); and to test whether mutations(s) in CFRDSP cause(s) either erd or prcd.
Methods: Three erd-affected dogs were crossbred to three prcd-affected dogs, and their progeny were tested by electroretinography and retinal morphology for evidence of retinal degeneration. Canine RDS/peripherin cDNA was cloned and sequenced after reverse-transcription-polymerase chain reaction (RT-PCR) of total retinal RNA. A set of overlapping fragments of CFRDSP cDNA amplified from normal and prcd-affected retinal RNA was examined by double-stranded conformational polymorphism analysis for evidence of any mutation in prcd-affected dogs. RDS/peripherin-specific restriction fragment length polymorphism (RFLP) allelic differences within informative prcd and erd pedigrees were sought by digestion of amplified regions of the CFRDSP gene with different restriction enzymes. A Hinf I RFLP was identified with alleles segregating in a set of prcd and erd informative pedigrees. Linkage of CFRDSP to either prcd or erd was tested using the LINKAGE analysis package.
Results: All progeny from the erd x prcd cross were phenotypically normal at ages beyond the age of diagnosis for both parental disorders. The sequence of CFRDSP cDNA is reported (Genbank accession U27349). It is, overall, 79% identical at the nucleotide level with the corresponding human sequence. The coding region shares 89% and 93% nucleotide identity with the corresponding human and feline sequences, respectively. No mutation has been identified in the coding region of CFRDSP in prcd-affected dogs. In prcd pedigrees informative for both prcd and the CFRDSP Hinf I RFLP, a minimum of six obligate recombinants were identified. Similarly, in erd pedigrees, 14 of 29 progeny informative for both erd and this RFLP were obligate recombinants.
Conclusions: The canine erd and prcd mutations are nonallelic. The canine RDS/peripherin gene (CFRDSP) has been excluded as a candidate for both prcd and erd. The demonstrated informativeness of the canine pedigrees on which these studies were based will enable testing other candidate genes for prcd and erd. Sequence information of CFRDSP will enable testing this locus as a candidate in other canine hereditary retinal degenerations.