Abstract
Purpose: :
To determine how arrestin availability affects the rate of recovery in rod photoreceptors.
Methods: :
We used two-flash ERG to determine the rate of rod recovery in vivo.
Results: :
Mice expressing arrestin at various levels: 4% (Tr-4Arr-/-), 12% (Tr-12Arr-/-), 50% (Arr+/-), and 100% of wild-type (WT), were tested by two-flash ERG with the first (desensitizing) flash at 160, 400, 1000, and 2500 photons/rod. The level of recovery was determined by the response to a probe flash of 4500 photons/rod delivered at variable intervals after the desensitizing flash. We showed that recovery in WT retinas progressively slows with increasing intensity of the initial flash. Time of half recovery is ~ 2.5-fold longer after the highest flash intensity (2500 photons/rod) than after the lowest (160 photons/rod). Mice expressing arrestin at 50% and 12% of the normal level were not statistically different from WT mice at any light intensity tested. Interestingly, the mice expressing the lowest level of arrestin (4%) had a dramatically slower recovery than the other three genotypes at all intensities. In these mice, the time of half recovery increased ~28 fold when tested with the highest flash intensity, in contrast to ~2.5 in other animals. Even after the dimmest desensitizing flash, the rate of recovery of Tr-4Arr-/- rods was two times slower than in other lines.
Conclusions: :
Our data show that arrestin expression between 100% and 12% of WT is sufficient for rapid recovery at different light levels. Gradual slowing of recovery with increasing intensity of the first flash observed in Tr-12Arr-/-, Arr+/-, and WT animals becomes dramatic with limited arrestin in Tr-4Arr-/- mice. This suggests that there is a threshold of arrestin expression between 4% and 12% of WT for transition from near normal to abnormally slow photoresponse recovery.
Keywords: photoreceptors • electroretinography: non-clinical • protein structure/function