From candelas to photoisomerizations in the mouse eye by rhodopsin bleaching in situ and the light-rearing dependence of the major components of the mouse ERG

Abstract

To quantify the rate at which light in a ganzfeld produces photoisomerizations in mouse rods in situ, we measured the rate of rhodopsin bleaching in eyes of recently euthanized mice with fully dilated pupils. The amount of rhodopsin declined as a first-order (exponential) function of the duration of the exposure at the luminance of 920 scot cd m −2: the rate constants of bleaching were 8.3 × 10 −6 and 2.8 × 10 −5 s −1 (scot cd −1 m 2) −1 for C57B1/6 and 129P3/J mice, respectively. When the ∼3-fold difference in effective areas of the pupils of the mice are taken into consideration, the bleaching rates for both strains become essentially the same, 2.6 × 10 −6 fraction rhodopsin (scot Td s) −1. Assuming 7 × 10 7 rhodopsin molecules per rod, this bleaching rate yields the result that a flash of 1 scot Td s produces 181 photoisomerizations per rod, a value close to that derived from analysis of the collecting area of the rod for axially propagating light. We measured the electroretinograms of mice of the two strains reared under controlled illumination conditions (2 and 100 lux), and compared their properties, using the calibrations to determine the absolute sensitivities of the b-wave and a-waves. The intensity that produces a half-saturating rod b-wave response is 0.3–0.6 photoisomerizations rod −1, and the amplification constant of the rod a-wave is 5–6 s −2 photoisomerization −1, with little dependence on the strain.