Vision begins when light is absorbed by visual pigments. It is commonly believed that the absorption spectra of visual pigments are modulated by interactions between the retinal and amino acids within or near 4.5 Å of the retinal in the transmembrane (TM) segments. However, this dogma has not been rigorously tested. In this study, we show that the retinal-opsin interactions extend well beyond the retinal binding pocket. We found that, although it is positioned outside of TM segments, the C-terminus of the rhodopsin in the rockfish longspine thornyhead (Sebastolobus altivelis) modulates its λmax by interacting mainly with the last TM segment. Our results illustrate how amino acids in the C-terminus are likely to interact with the retinal. We anticipate our analyses to be a starting point for viewing the spectral tuning of visual pigments as interactions between the retinal and key amino acids that are distributed throughout the entire pigment.
Crystallins in the retina may serve a chaperone-like protective function. In this study we measured mRNA levels for alpha-, beta- and gamma-crystallins in rat retinas following treatment with potentially damaging levels of light. We also determined crystallin protein patterns in photoreceptor cell rod outer segments (ROSs) isolated from rats exposed to intense light. Weanling albino rats were maintained in a dim cyclic light environment or in darkness for 40 days. At P60 animals were treated with intense visible light, for as long as 8 h, beginning at various times of the day or night. Retinas were excised immediately after light treatment and used for quantitative RT-PCR, or to prepare ROSs for western analysis. Some eyes were frozen in OCT for crystallin immunohistochemistry. Intense light exposure led to increases in mRNA expression for all retinal crystallins and to changes in ROS crystallin immunoreactivity. These light-induced changes were found to depend on the time of day that exposure started, duration of light treatment and previous light rearing history. We suggest that crystallin synthesis in retina exhibits a dependence on both light stress and circadian rhythm and that within photoreceptor cells crystallins appear to migrate in a light-independent, circadian fashion.