Availability of 11-cis retinal and opsins without chromophore as revealed by small bleaches of rhodopsin in excised albino mouse eyes

Small bleaches were used to study the rhodopsin regeneration process. At bleaches from 5.2% to 24.7%, the rhodopsin regenerations were consistent with a one-for-one recovery of bleached molecules. At response saturation rod photoreceptors exhibit a bleach level of only 5%. Major increases in rhodopsin regeneration were observed at bleach levels between 1.3% and 5.2%. The rhodopsin regenerations exhibited a linear relationship that was 4-times the bleach (dark adaptations of 0.75 and 1.5 h). The data show that the bleach initiates the availability, and possibly production, of 11-cis retinal in amounts that are 4-times the number of bleached molecules within the functional range of the rod photoreceptors. Rhodopsin regeneration also requires the presence of opsins without chromophore. Regenerations beyond the bleach indicate the presence of such opsins prior to the bleach. The opsin amounts were 8.1%, 8.6%, 3.1% and 0% of the total visual pigment at dark adaptation times of 0.75, 1.5, 24 and 48 h, respectively. Those opsins, as well as the ones produced by the bleach, may be regenerated to rhodopsin following a small bleach or with additional time in the dark.