A mammalian-like melanopsin (Opn4m) has been found in all major vertebrate classes except reptile. Since the pupillary light reflex (PLR) of the fresh water turtle takes between 5 and 10 min to achieve maximum constriction, and since photosensitive retinal ganglion cells (ipRGCs) in mammals use Opn4m to control their slow sustained pupil responses, we hypothesized that a Opn4m homolog exists in the retina of the turtle. To identify its presence, retinal tissue was dissected from seven turtles, and total RNA extracted. Reverse transcriptase-polymerase chain reactions (RT-PCRs) were carried out to amplify gene sequences using primers targeting the highly conserved core region of Opn4m, and PCR products were analyzed by gel electrophoresis and sequenced. Sequences derived from a 1004-bp PCR product were compared to those stored in GenBank by the basic local alignment search tool (BLAST) algorithm and returned significant matches to several Opn4ms from other vertebrates including chicken. Quantitative real-time PCR (qPCR) was also carried out to compare expression levels of Opn4m in different tissues. The normalized expression level of Opn4m in the retina was higher in comparison to other tissue types: iris, liver, lung, and skeletal muscle. The results suggest that Opn4m exists in the retina of the turtle and provides a possible explanation for the presence of a slow PLR. The turtle is likely to be a useful model for further understanding the photoreceptive mechanisms in the retina which control the dynamics of the PLR.

Our goal in this study was to examine the red-eared slider turtle for a photomechanical response (PMR) and define its spectral sensitivity. Pupils of enucleated eyes constricted to light by ∼11%, which was one-third the response measured in alert behaving turtles at ∼33%. Rates of constriction in enucleated eyes that were measured by time constants (1.44-3.70 min) were similar to those measured in turtles at 1.97 min. Dilation recovery rates during dark adaptation for enucleated eyes were predicted using line equations and computed times for reaching maximum sizes between 26 and 44 min. Times were comparable to the measures in turtles where maximum pupil size occurred within 40 min and possessed a time constant of 12.78 min. Hill equations were used to derive irradiance threshold values from enucleated hemisected eyes and then plot a spectral sensitivity curve. The analysis of the slopes and maximum responses revealed contribution from at least two different photopigments, one with a peak at 410 nm and another with a peak at 480 nm. Fits by template equations suggest that contractions are triggered by multiple photopigments in the iris including an opsin-based visual pigment and some other novel photopigment, or a cryptochrome with an absorbance spectrum significantly different from that used in our model. In addition to being regulated by retinal feedback via parasympathetic nervous pathways, the results support that the iris musculature is photointrinsically responsive. In the turtle, the control of its direct pupillary light response (dPLR) includes photoreceptive mechanisms occurring both in its iris and in its retina.

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