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S. Hotop, J. Beatrice, A. Wenzel, C.E. Remé, C. Grimm; RPE65 Does Not Account for Increased Light Damage Susceptibility at Night in Rats . Invest. Ophthalmol. Vis. Sci. 2003;44(13):5131.
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Purpose: The susceptibility of rats to light-induced retinal degeneration is increased at night. In mice, an important determinant of light damage susceptibility is the efficacy of rhodopsin regeneration after bleaching. The rate of rhodopsin regeneration is at least partly controlled by RPE65, a protein expressed in the retinal pigment epithelium. We therefore tested a potential involvement of RPE65 and rhodopsin regeneration in the increased light damage susceptibility of rats at night. Methods: Rats were exposed to visible light at noon or at midnight and extent of light damage was determined by retinal morphology and TUNEL staining. Rpe65 gene expression was analyzed by semiquantitative RT-PCR and levels of RPE65 protein were determined by Western blotting. Rhodopsin regeneration kinetics was determined by measuring rhodopsin content immediately after a strong bleach and after different times of recovery in darkness. Results: Rats were more susceptible to light damage at night as described by Organisciak and collegues (Invest. Ophthalmol. Vis. Sci. 2000,41,3694). Rpe65 gene expression followed a day – night rhythm with highest steady-state mRNA levels at the beginning and lowest levels at the end of the day period. However, RPE65 protein levels remained constant throughout the day. Rhodopsin regeneration kinetics did not differ during day and night. Conclusions: Levels of RPE65 protein and rhodopsin regeneration kinetics do not correlate with the increased light damage susceptibility observed in rats at night. Additional genetic or physiologic modifiers may exist in rats that regulate the retinal responsiveness to acute light exposure.
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