Purchase this article with an account.
A. McKee, M. Ader, A. Kennan, P.F. Kenna, G. Farrar, P. Humphries; Overexpression of A20 Induces Aberrant Retinal Morphology . Invest. Ophthalmol. Vis. Sci. 2005;46(13):1684.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: We recently identified several transcripts showing significantly altered expression in the retinas of mice with a targeted disruption of the rhodopsin gene. Rho–/– mice do not elaborate rod outer segments, losing their photoreceptors by apoptosis over 3 months. A20, a known antiapoptotic gene and potent inhibitor of nuclear factor kappa B (NF–kB) was shown to be considerably downregulated in the knockout retina. Using the technique of in vivo and in vitro retinal electroporation we have introduced an A20 plasmid into retinas of Rho–/– mice to analyze its function and investigate its potential role in retinal development and disease. Methods: A CMV driven A20–GFP plasmid was directly injected into the subretinal space of knockout and wild type neonatal mouse pups and electric pulses were subsequently applied using tweezer electrodes. Electroretinograph (ERG) responses were measured 6 weeks post injection. Eyes were then enucleated and sectioned for histological analysis. A similar technique was used for in vitro electroporation of retinal explants using a micro electroporation chamber. These explants were then cultured for two weeks, sectioned using a vibratome and analyzed by immunocytochemistry. Results: ERG rod responses in Rho–/– eyes showed little sign of improvement 6 weeks post injection in comparison with uninjected controls. Histological and immunocytological sections of whole eyes and retinal explants showed that retinas overexpressing A20 had considerable morphological differences compared to sham injected eyes, uninjected eyes, or control retinas transfected with a GFP expressing plasmid. Conclusions: Because A20 is antiapoptotic and is known to inhibit NF–kB it is likely that the unusual phenotype observed in retinas overexpressing this gene is due to altered apoptosis. It is also interesting to speculate that NF–kB may play a role in retinal development
This PDF is available to Subscribers Only