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S. S. Lee, D. R. Hinton, L. Chan, C. Spee, P. Sreekumar, G. J. Chader, J. D. Weiland; The Role of Alpha Crystallins on Safety of Acute Electrical Retinal Stimulation in Mice. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3053.
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The purpose of this study is to evaluate the role of retinal α-crystallins, members of the heat shock family of proteins, on the safety and effectiveness of acute electrical retinal stimulation in mice.
The experiment involved temporary placement of a stimulating electrode on the cornea of mice. The electrode was held in place by an articulating arm for the duration of the procedure. Groups of 2-3 month old mice (4 αA-crystallin knock out (KO) mice, 3 αB-crystallin KO; and 10 129S6/SvEvTac control mice) underwent acute stimulation for 1 hour (0, 250, or 500 µA at 2 Hz) and sacrificed after 3, 7, or 14 days. Enucleated eyes were bisected with posterior portions used for Hematoxylin and Eosin (H&E) and immunohistochemical staining. Sections stained for α-crystallins and glial fibrillary acidic protein (GFAP) were imaged using a Zeiss LSM510 confocal microscope.
Preliminary results show increased crystallin expression was observed in 129S6/SvEvTac mice that underwent extraocular electrical stimulation. Alpha-A crystallin staining was the most prominent and widespread. The areas that generally showed the highest amount of immunopositivity were the ganglion cell and retinal pigment epithelial layers (RPE). Both the 129S6/SvEvTac and αA-crystallin KO mice that had undergone stimulation showed no change in H&E morphology and were virtually identical to wild type retinas. Only αB-crystallin KO mice showed any significant histological changes after stimulation. In all cases with αB-crystallin KO mice, cellular layer disruption was observed at the ciliary margins. In one case, exudate and RPE hypertrophy was also present in the posterior retina. Higher current levels did show GFAP upregulation at the ciliary margins across all groups.
These results suggest that the retina is highly tolerant of electrical stimulation and that alpha-B crystallins may play a role in its response to injury.
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