Distribution and axial diffusion of retinol in bleached rod outer segments of frogs (Rana pipiens).

Isolated retinas and rod outer segments from frogs (Rana pipiens) were exposed to light that produced axially uniform total bleaches of rhodopsin. Using fluorescence video microscopy, it was shown that the formation and equilibrium distribution of all-trans-retinol, the final chromophore product of rhodopsin bleaching is axially uniform. This result shows that the rate and amount of oxidoreductase-mediated reduction of all-trans-retinal to all-trans-retinol is not affected by the relative age of the disk membranes to which the enzymes are bound. Therefore previously reported axial differences in regeneration of rhodopsin and recovery of photocurrent after exposure to bright light probably are not due to axial differences in the formation of rhodopsin photoproducts. In addition, measurements on individual rod cells show that there is no significant redistribution of retinol for up to 2 hr following localized partial bleaches of rhodopsin. This raises the perplexing question of how retinol is shuttled between disk membranes and the pigment epithelium during visual pigment regeneration following substantial bleaches.