Antisense Inactivation of Rds/Peripherin in Xenopus Laevis Embryonic Retinal Cultures

Abstract

To investigate the role of rds/J.peripherin in photoreceptor outer segment membrane assembly, we have transfected embryonic Xenopus laevis retinas with phosphorothioated antisense or sense RNA complementary to specific regions of the three Xenopus rds/peripherin homologs. We have demonstrated that using antisense oligonucleotides complementary to the three homologs of rds/peripherin, we can significantly alter the structure of photoreceptor outer and inner segments. In addition, the immunolocalization patterns using xrds35 and -38 anti-peptide sera and an anti-opsin monoclonal antibody were significantly reduced, suggestive of aberrant outer segment membrane assembly. We have successfully generated a model upon which to study the functional role of rds/peripherin, the product of the rds gene, by down-regulating the expression of this protein in developing photoreceptors. A better understanding of rds/peripherin gene expression will shed light on the role of this molecule in rod and cone photoreceptor outer segment development, structure, function and survival. This will also provide additional insights into the mechanisms of, and possibly open new therapeutic avenues for, the retinal degenerative processes taking place in the heterogeneous clinical patterns so far described in association with peripherinJ.RDS human gene mutations in association with autosomal dominant retinitis pigmentosa.