Journal
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
Volume 45, Issue 8, Pages 2514-2521Publisher
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
DOI: 10.1167/iovs.04-0065
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Funding
- NEI NIH HHS [R56 EY010609, R01 EY007361-11, R01 EY010609-09, R01 EY007361, R01 EY010609-08, EY-07361, R01 EY010609, EY-10609, P30 EY012190] Funding Source: Medline
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PURPOSE. Mutations in the photoreceptor-specific protein peripherin/rds are associated with multiple retinal diseases. To date, attempts to achieve complete structural and functional rescue in animal models of peripherin/rds-induced retinal degeneration have not been successful. Gene therapy-directed approaches have been hindered by the haploinsufficiency phenotype, which dictates well-regulated expression of peripherin/rds protein levels. METHODS. Using a transgenic mouse line expressing wildtype peripherin/rds (NMP), the authors evaluated the critical in vivo level of peripherin/rds needed to maintain photoreceptor structure and ERG function and assessed the consequences of peripherin/rds overexpression in both rods and cones by Western blot and immunoprecipitation analyses, immunohistochemistry, electron microscopy, and electroretinography. The NMP transgene included a C-terminal modification (P341Q) to facilitate detection of the transgenic protein in the presence of wild-type peripherin/rds, using the monoclonal antibody 3B6. RESULTS. Peripherin/rds protein levels in NMP homozygotes were similar to60% of wild-type levels. Western blot and immunoprecipitation analyses confirmed normal biochemical properties of the NMP protein when compared with wild-type peripherin/rds. Immunohistochemistry demonstrated appropriate localization of transgenic peripherin/rds protein to the disc rim region of photoreceptor outer segments. Total peripherin/rds levels in the retina were modulated by crossing NMP transgenic mice into different rds genetic backgrounds. A positive correlation was observed between peripherin/rds expression levels and the structural and functional integrity of photoreceptor outer segments. Overexpression of peripherin/rds caused no detectable adverse effects on rod or cone structure and function. CONCLUSIONS. These findings may have significant implications regarding therapeutic intervention in peripherin/rds-associated retinal diseases.
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