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Restoration of photoreceptor ultrastructure and function in retinal degeneration slow mice by gene therapy

Abstract

The gene Prph2 encodes a photoreceptor-specific membrane glycoprotein1, peripherin-2 (also known as peripherin/rds), which is inserted into the rims of photoreceptor outer segment discs in a complex with rom-1 (ref. 2). The complex is necessary for the stabilization of the discs, which are renewed constantly throughout life, and which contain the visual pigments necessary for photon capture3. Mutations in Prph2 have been shown to result in a variety of photoreceptor dystrophies, including autosomal dominant retinitis pigmentosa and macular dystrophy4. A common feature of these diseases is the loss of photoreceptor function, also seen in the retinal degeneration slow (rds or Prph2 Rd2/Rd2) mouse, which is homozygous for a null mutation in Prph2. It is characterized by a complete failure to develop photoreceptor discs and outer segments5, downregulation of rhodopsin6,7 and apoptotic loss of photoreceptor cells8,9. The electroretinograms (ERGs) of Prph2Rd2/Rd2 mice have greatly diminished a-wave and b-wave amplitudes, which decline to virtually undetectable concentrations by two months10. Subretinal injection of recombinant adeno-associated virus (AAV) encoding a Prph2 transgene results in stable generation of outer segment structures and formation of new stacks of discs containing both perpherin-2 and rhodopsin, which in many cases are morphologically similar to normal outer segments. Moreover, the re-establishment of the structural integrity of the photoreceptor layer also results in electrophysiological correction. These studies demonstrate for the first time that a complex ultrastructural cell defect can be corrected both morphologically and functionally by in vivo gene transfer.

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Figure 1: Immunohistochemical detection of peripherin-2 in a Prph2Rd2/Rd2 mouse 3 weeks after subretinal injection of AAV.rho.prph2.
Figure 2: Comparison of peripherin-2 expression in Prph2Rd2/Rd2 mice 3 and 6 weeks after subretinal injection of AAV.rho.prph2.
Figure 3: Co-localization of peripherin-2 and rhodopsin in Prph2Rd2/Rd2 mice 3 weeks after subretinal injection of AAV.rho.prph2.
Figure 4: TEM analysis of Prph2Rd2/Rd2 mouse retina 4 weeks after subretinal injection of AAV.rho.prph2.
Figure 5: SEM of the outer surface of the Prph2Rd2/Rd2 mouse neuroretina 6 weeks after subretinal injection of AAV.rho.prph2.
Figure 6: Representative ERG traces.

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Acknowledgements

We thank G. Holder, C. Hogg and M. Seeliger for advice on ERG and X. Zhang for supplying the original vector plasmids. This work was supported by grants from the Foundation Fighting Blindness (USA), the Wellcome Trust, Fight for Sight, and DAAD/British Council. Personal support for GS from Ausbildungsstiftung für den Kanton Schwyz; EMDO Stiftung, Schweiz. Fonds zurVerhütung und Bekämpfung der Blindheit and Holderbank-Stiftung.

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Correspondence to Robin R. Ali.

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Ali, R., Sarra, GM., Stephens, C. et al. Restoration of photoreceptor ultrastructure and function in retinal degeneration slow mice by gene therapy. Nat Genet 25, 306–310 (2000). https://doi.org/10.1038/77068

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