Exceptional Visual-Opsin Coexpression and Phenotypic Diversity in Outer-Retinal Photoreceptors of Caenophidian Snakes.

Abstract

Snakes are a valuable yet understudied taxon for investigating evolutionary adaptations in the vertebrate retina. They possess up to three visual pigments: a short-wavelength-sensitive opsin (SWS1), a medium/long-wavelength-sensitive opsin (LWS), and rhodopsin (RH1). Nocturnal snakes have duplex retinas containing both rod and cone photoreceptors, whereas diurnal caenophidian ("advanced") snakes exhibit simplex "all-cone" retinas, lacking morphologically typical rods. In this study, we analyzed photoreceptor morphology in the retinas of caenophidian snakes using high-resolution scanning electron microscopy (SEM) and examined visual-opsin expression patterns with immunohistochemistry (IHC). Our analyses revealed remarkable interspecific variability in visual-cell morphology. Light microscopy showed that in all sampled diurnal caenophidians, photoreceptors expressing RH1 exhibit a gross cone-like morphology. However, SEM analysis revealed a subset of photoreceptors with distinct features-thinner inner segments and rod-like synaptic terminals-suggesting they are transmuted, cone-like rods. In retinal sections from nocturnal caenophidian snakes, coexpression of the cone opsins SWS1 and LWS in individual cones was observed, whereas rhodopsin expression remained restricted to morphologically typical rods and showed no coexpression. In contrast, diurnal caenophidians commonly coexpress rhodopsin and SWS1 in single cones, with some instances of triple coexpression (SWS1, RH1, and LWS) in single cones. We evaluated the patterns of spatial distribution of RH1- and SWS1-expressing photoreceptors, as well as SWS1 + RH1 multiopsin cones, in wholemounted retinas of ten species. Our findings revealed considerable species-specific variation in photoreceptor density, topography, and opsin coexpression patterns. IHC results suggest that in some species, rhodopsin is not only expressed in transmuted, cone-like rods but may also be co-opted by UV/violet-sensitive (SWS1-expressing) cones. These findings underscore the exceptional diversity and adaptive innovation in snake visual systems. The unique features and striking interspecific differences in their photoreceptors highlight snakes as an outstanding taxon for studying vertebrate visual-system function and evolution.