Cone Pigments Research Articles

Molecular Mechanisms Underlying the Spectral Shift in Zebrafish Cone Opsins.

Visual pigments are essential for converting light into electrical signals during vision. Composed of an opsin protein and a retinal-based chromophore, pigments in vertebrate rods (Rh1) and cones (Rh2) have different spectral sensitivities, with distinct peak absorption wavelengths determined by the shape and composition of the chromophore binding pocket. Despite advances in understanding Rh1 pigments such as bovine rhodopsin, the molecular basis of spectral shifts in Rh2 cone opsins has been less studied, particularly the E122Q mutation, which accounts for about half of the observed spectral shift in these pigments. In this study, we employed molecular modeling and quantum mechanical techniques to investigate the molecular mechanisms behind the spectral difference in blue-shifted Rh2-1 (absorption peak = 467 nm, 122Q) and green-shifted Rh2-4 (absorption peak = 505 nm, 122E) zebrafish cone opsins. We modeled the pigments 3D structures based on their sequences and conducted all-atom molecular dynamics simulations totaling 2 microseconds. Distance analysis of the trajectories identified three key sites: E113, E181, and E122. The E122Q mutation, previously known, validates our findings, while E181 and E113 are newly identified contributors. Structural analysis revealed key features with differing values that explain the divergent spectral sensitivities of Rh2-1 and Rh2-4: 1) chromophore atom fluctuations and C5-C6 torsion angle, 2) binding pocket volume, 3) hydration patterns, and 4) E113-chromophore interaction stability. Quantum mechanics further confirms the critical role of residue E181 in Rh2-1 and E122 in Rh2-4 for their spectral behavior. Our study provides new insights into the molecular determinants of spectral shifts in cone opsins, and we anticipate that it will serve as a starting point for a broader understanding of the functional diversity of visual pigments.

Dominant role for pigment epithelial CRALBP in supplying visual chromophore to photoreceptors

Cellular retinaldehyde-binding protein (CRALBP) supports production of 11-cis-retinaldehyde and its delivery to photoreceptors. It is found in the retinal pigment epithelium (RPE) and Müller glia (MG), but the relative functional importance of these two cellular pools is debated. Here, we report RPE- and MG-specific CRALBP knockout (KO) mice and examine their photoreceptor and visual cycle function. Bulk visual chromophore regeneration in RPE-KO mice is 15-fold slower than in controls, accounting for their delayed rod dark adaptation and protection against retinal phototoxicity, whereas MG-KO mice have normal bulk visual chromophore regeneration and retinal light damage susceptibility. Cone pigment regeneration is significantly impaired in RPE-KO mice but mildly affected in MG-KO mice, disclosing an unexpectedly strong reliance of cone photoreceptors on the RPE-based visual cycle. These data reveal a dominant role for RPE-CRALBP in supporting rod and cone function and highlight the importance of RPE cell targeting for CRALBP gene therapies.

Visual pigment concentration and photoreceptor outer segment length in the human retina.

The Beer-Lambert law suggests that visual pigment optical density (OD) should be linearly related to the length of photoreceptor outer segments (POSs). Mammalian studies indicate that visual pigment concentration increases with POS length, but the nature of this relationship may vary due to factors such as visual pigment packing density or retinal eccentricity, and may not necessarily be linearly related. The purpose of this study was to establish the relationship between OD and POS length in humans. Spectral domain optical coherence tomography (OCT) was used to image POS, and imaging retinal densitometry (IRD) was used to measure OD at corresponding locations in 19 healthy participants (age range 25-82 years). POS length and OD measurements were extracted from OCT and IRD images at 23 discrete locations spanning the central 9° of the retina. The averaged data from all participants were fitted with models based on the Beer-Lambert law to establish the relationship between OD and POS length. Visual pigment OD increased monotonically with POS length, but the relationship was non-linear, and a straight-line fit, based on a simple interpretation of the Beer-Lambert law, provided a poor description. A model allowing for different rod and cone visual pigment concentrations provided a superior fit. Specifically, the data were well described by a model where the molar concentration of visual pigment in cones and rods were 3.8 × 10-3 mol/L and 1.8 × 10-3mol/L, respectively. In accordance with the Beer-Lambert law, the results indicate that OD increases monotonically with POS length in humans, but the precise relationship is dependent on photoreceptor type. These results suggest that visual pigment concentration in rods is only about 48% of that found in cones. This may be due to the ubiquitous nature of artificial light that works to reduce the concentration of rhodopsin in rod photoreceptors.

Convergent mechanism underlying the acquisition of vertebrate scotopic vision

High sensitivity of scotopic vision (vision in dim light conditions) is achieved by the rods’ low background noise, which is attributed to a much lower thermal activation rate (kth) of rhodopsin compared with cone pigments. Frogs and nocturnal geckos uniquely possess atypical rods containing noncanonical cone pigments that exhibit low kth, mimicking rhodopsin. Here, we investigated the convergent mechanism underlying the low kth of rhodopsins and noncanonical cone pigments. Our biochemical analysis revealed that the kth of canonical cone pigments depends on their absorption maximum (λmax). However, rhodopsin and noncanonical cone pigments showed a substantially lower kth than predicted from the λmax dependency. Given that the λmax is inversely proportional to the activation energy of the pigments in the Hinshelwood distribution-based model, our findings suggest that rhodopsin and noncanonical cone pigments have convergently acquired low frequency of spontaneous-activation attempts, including thermal fluctuations of the protein moiety, in the molecular evolutionary processes from canonical cone pigments, which contributes to highly sensitive scotopic vision.

Glu1022.53-Mediated Early Conformational Changes in the Process of Light-Induced Green Cone Pigment Activation.

Ligand-triggered activation of G protein-coupled receptors (GPCRs) relies on the phenomenon of loose allosteric coupling, which involves conformational alterations spanning from the extracellular ligand-binding domain to the cytoplasmic region, where interactions with G proteins occur. During the GPCR activation process, several intermediate and equilibrium states orchestrate the movement of the flexible and rigid transmembrane (TM) segments of the GPCR. Monitoring early conformational changes is important in unraveling the structural intricacies of the loose allosteric coupling. Here, we focus on the lumi intermediate formed by thermal relaxation from the initial photointermediate, batho in primate green cone pigment (MG), a light-sensitive GPCR responsible for color vision. Our findings from light-induced Fourier transform infrared difference spectroscopy reveal its similarity with rhodopsin, which mediates twilight vision, specifically involving the flip motion of the β-ionone ring, the relaxation of the torsional structure of the retinal, and local perturbations in the α-helix upon lumi intermediate formation. Conversely, we observe a hydrogen bond modification specific to MG's protonated carboxylic acid, identifying its origin as Glu1022.53 situated in TM2. The weakening of the hydrogen bond strength at Glu1022.53 during the transition from the batho to the lumi intermediates corresponds to a slight outward movement of TM2. Additionally, within the X-ray crystal structure of the rhodopsin lumi intermediate, we note the relocation of the Met862.53 side chain in TM2, expanding the volume of the retinal binding pocket. Consequently, the position of 2.53 emerges as the early step in the conformational shift toward light-induced activation. Moreover, given the prevalence of IR-insensitive hydrophobic amino acids at position 2.53 in many rhodopsin-like GPCRs, including rhodopsin, the hydrogen bond alteration in the C═O stretching band at Glu1022.53 of MG can be used as a probe for tracing conformational changes during the GPCR activation process.

Effect of pre‐receptor filters on chromatic sensitivity and other visual functions in the eye

Spatial and spectral information carried by light emitted from visual displays is filtered as it passes through spectacle lenses in front of the eye as well as by filters within the eye. The image forming and the spectral transmittance properties of the elements involved in image formation in the eye cause changes in both the luminance and colour contrast in retinal images. In addition, the amount of light that reaches the retina is always reduced by pre‐receptor filters and this reduction in retinal illuminance can cause diminished retinal sensitivity to both luminance and colour contrast. Of particular interest are the use of coloured lenses in front of the eye to reduce the short‐wavelength content of the light that reaches the retina and the use of ‘notch’ filters to absorb light selectively in certain wavebands, with inevitable consequences on both the luminance and chromatic contrast of retinal images. The presence of variant cone‐pigments in some subjects or the complete absence of one class of cone pigment in others, can also produce large deviations in both luminance and colour contrast signals that can either enhance or diminish the conspicuity of coloured objects (doi: 10.1364/josaa.22.000017).The purpose of this study was to develop and validate a model of red/green (RG) and yellow/blue (YB) colour discrimination that can also predict how chromatic sensitivity is affected by the use of spectrally‐selective, spectacle lenses or by filters internal to the eye, when viewing visual displays. We examined how RG and YB colour thresholds relate to the corresponding cone contrasts needed for detection of colour differences. The results reveal an invariance of cone contrasts at threshold for different states of chromatic adaptation. In other words, colour contrast signals at threshold follow Weber's law (doi: 10.1111/j.1475‐1313.2010.00773.x). A combination of RG and YB colour signals is needed to predict the colour threshold ellipse. The model relies on these findings and predicts how coloured filters external to the eye as well as pre‐receptor filters within the eye alter the measured RG and YB colour thresholds. Model predictions have been compared against measured colour thresholds using the CAD test (doi: 10.1093/bmb/ldx007). A number of experiments were carried out when the visual display was viewed with no lens in front of the eye or through a number of tinted lenses designed to absorb progressively more blue light.The effects of pre‐receptor filters within the eye such as the lens and the macular pigment have also been investigated. The results show that ‘blue‐blocking’ filters employed in typical spectacle lenses used with daylight illumination cause only small shifts, approximately along the daylight locus, without any significant loss of either RG or YB chromatic sensitivity. ‘Orange’ lenses that absorb greater amounts of short‐wavelength light cause a significant increase in YB thresholds, but have little or no effect on RG thresholds. More surprisingly, the model also predicts that the optical density of the macular pigment has little or no effect on the size of the colour threshold ellipse.Keywords: colour, cone contrasts, blue light, CAD test.

Molecular Mechanism of Spectral Tuning by Chloride Binding in Monkey Green Sensitive Visual Pigment

The visual pigments of the cones perceive red, green, and blue colors. The monkey green (MG) pigment possesses a unique Cl- binding site; however, its relationship to the spectral tuning in green pigments remains elusive. Recently, FTIR spectroscopy revealed the characteristic structural modifications of the retinal binding site by Cl- binding. Herein, we report the computational structural modeling of MG pigments and quantum-chemical simulation to investigate its spectral redshift and physicochemical relevance when Cl- is present. Our protein structures reflect the previously suggested structural changes. AlphaFold2 failed to predict these structural changes. Excited-state calculations successfully reproduced the experimental red-shifted absorption energies, corroborating our protein structures. Electrostatic energy decomposition revealed that the redshift results from the His197 protonation state and conformations of Glu129, Ser202, and Ala308; however, Cl- itself contributes to the blueshift. Site-directed mutagenesis supported our analysis. These modeled structures may provide a valuable foundation for studying cone pigments.

Impact of matching field size on color matching (functions) accuracy

AbstractAccurate colorimetry starts with accurate color matching functions (CMFs). Due to changes in the macular pigment and cone pigment optical densities at different retinal locations, different CMFs are required for different stimulus field sizes. To characterize the accuracy of the CIE recommendation for the use of 2° and 10° standard CMFs and the field‐size dependent CIEPO06 model, in this study, a series of achromatic matching experiments were performed with 2°, 4°, 6°, 8°, and 10° bipartite fields using spectrally narrowband primaries. Using the CIEPO06 model, optimal field sizes were estimated that minimize the chromaticity differences between the spectrally narrowband observer matches and the spectrally broadband achromatic target. It was found that the optimal field size estimated using the CIEPO06 model is close to half the actual bipartite field size in most cases, except for the 2° field. The discrepancy between the 10° bipartite field in Stiles & Burch's experiments and our optimal field size (6.54°) was assumed to be due to different individual color comparison strategies. However, the results of experiments requiring specific observer gaze positions did not support this assumption and the reason for the discrepancy remains unclear. Confirming our earlier results, the primary set (636, 521, 447 nm) was again found to be largely insensitive to changes in CMFs and to provide the most accurate matches under various fields of view. The inter‐ and intra‐observer variability ellipses for 2° matches are larger than those for 10°, consistent with Brown's finding for color discrimination ellipses. The magnitude of the intraobserver variability was similar for all field sizes, except for 2° field size, where matching errors were larger for some primary sets.

Grk7 but not Grk1 undergoes cAMP-dependent phosphorylation in zebrafish cone photoreceptors and mediates cone photoresponse recovery to elevated cAMP

In the vertebrate retina, phosphorylation of photoactivated visual pigments in rods and cones by G protein-coupled receptor kinases (GRKs) is essential for sustained visual function. Previous invitro analysis demonstrated that GRK1 and GRK7 are phosphorylated by PKA, resulting in a reduced capacity to phosphorylate rhodopsin. Invivo observations revealed that GRK phosphorylation occurs in the dark and is cAMP dependent. In many vertebrates, including humans and zebrafish, GRK1 is expressed in both rods and cones while GRK7 is expressed only in cones. However, mice express only GRK1 in both rods and cones and lack GRK7. We recently generated a mutation in Grk1 that deletes the phosphorylation site, Ser21. This mutant demonstrated delayed dark adaptation in mouse rods but not in cones invivo, suggesting GRK1 may serve a different role depending upon the photoreceptor cell type in which it is expressed. Here, zebrafish were selected to evaluate the role of cAMP-dependent GRK phosphorylation in cone photoreceptor recovery. Electroretinogram analyses of larvae treated with forskolin show that elevated intracellular cAMP significantly decreases recovery of the cone photoresponse, which is mediated by Grk7a rather than Grk1b. Using a cone-specific dominant negative PKA transgene, we show for the first time that PKA is required for Grk7a phosphorylation invivo. Lastly, immunoblot analyses of rod grk1a-/- and cone grk1b-/- zebrafish and Nrl-/- mouse show that cone-expressed Grk1 does not undergo cAMP-dependent phosphorylation invivo. These results provide a better understanding of the function of Grk phosphorylation relative to cone adaptation and recovery.

Human cone elongation responses can be explained by photoactivated cone opsin and membrane swelling and osmotic response to phosphate produced by RGS9-catalyzed GTPase

Human cone outer segment (COS) length changes in response to stimuli bleaching up to 99% of L- and M-cone opsins were measured with high resolution, phase-resolved optical coherence tomography (OCT). Responses comprised a fast phase (∼5 ms), during which COSs shrink, and two slower phases (1.5 s), during which COSs elongate. The slower components saturated in amplitude (∼425 nm) and initial rate (∼3 nm ms-1) and are well described over the 200-fold bleaching range as the sum of two exponentially rising functions with time constants of 80 to 90 ms (component 1) and 1,000 to 1,250 ms (component 2). Measurements with adaptive optics reflection densitometry revealed component 2 to be linearly related to cone pigment bleaching, and the hypothesis is proposed that it arises from cone opsin and disk membrane swelling triggered by isomerization and rate-limited by chromophore hydrolysis and its reduction to membrane-localized all-trans retinol. The light sensitivity and kinetics of component 1 suggested that the underlying mechanism is an osmotic response to an amplified soluble by-product of phototransduction. The hypotheses that component 1 corresponds to G-protein subunits dissociating from the membrane, metabolites of cyclic guanosine monophosphate (cGMP) hydrolysis, or by-products of activated guanylate cyclase are rejected, while the hypothesis that it corresponds to phosphate produced by regulator of G-protein signaling 9 (RGS9)-catalyzed hydrolysis of guanosine triphosphate (GTP) in G protein-phosphodiesterase complexes was found to be consistent with the results. These results provide a basis for the assessment with optoretinography of phototransduction in individual cone photoreceptors in health and during disease progression and therapeutic interventions.

Assessment of photoreceptor function with ultrafast retinal densitometry.

The optical density of visual pigment can be measured by imaging the dark-adapted eye while bleaching with visible light. This measurement can be made for individual photoreceptor cells using adaptive optics; however, activation of the phototransduction cascade imparts rapid changes in phase that modulate the signal via optical interference. This limits utility because data must be averaged over many experimental runs. Here we used a "flood" illuminated adaptive optics system at 4000 fps, bright light to achieve rapid bleaching, and broad illumination bandwidth to mitigate interference effects. Data were super-resolved using the natural motion of the eye to overcome the reduced pixel resolution of the ultrafast camera. This approach was applied to classify the trichromatic cone photoreceptor mosaic at a single fixation locus within the foveal region of 3 healthy subjects. Subjects were dark adapted for 6 minutes to replenish cone photopigment. This was followed either directly by imaging at 555 ± 50 nm, or by first pre-adapting the retina to 700 nm light to preferentially deplete "L" cone pigment. A total of 3,252 cones were classified as either "S", "M", or "L" type based on clustering of the intensity data observed under these two conditions. Mean classification probability ranged from 99.3 to 99.8%, with individual cell probabilities exceeding 95% in 97.0 to 99.2% of cones. Accuracy of cone classification peaked when using the first 10-30 ms of data, with significant reductions in accuracy noted with the inclusion of data from later times. Our results show that rapid bleaching and data acquisition significantly improve the robustness of cell-resolved densitometry.

Visual system plasticity is differently mediated by cone opsin expression and chromophore composition in closely related cichlid species

Phenotypic plasticity allows organisms to rapidly adjust to environmental changes. Cichlid fish inhabit a wide range of light environments and show a large diversity in visual system properties, which makes them a good model system to address the role of phenotypic plasticity in visual adaptation. Cichlid retinal cone pigments consist of opsin proteins bound to Vitamin A1 or A2-derived chromophores. Plasticity in expression has been shown for cichlid opsin genes, but less is known about the contribution of cyp27c1, the enzyme that converts Vitamin A1 into A2,. Here, we studied both opsin and cyp27c1 expression patterns for three closely related cichlid species from different visual habitats in Lake Victoria, across different light treatments. We found differences in cyp27c1 as well as in opsin expression patterns between the three species. Experimental light treatments affected the developmental trajectory of cyp27c1 expression in one species and opsin expression in all three species. Within each species, we found large individual variation in cyp27c1 expression levels and no consistent association with opsin expression levels. These results indicate that visual system plasticity of even closely related species can be differentially mediated by opsin and cyp27c1 expression, possibly associated with species differences in visual niche.

Comparison of Bovine and Carp Fish Visual Pigment Photo-Intermediates at Room Temperature.

This paper presents room temperature nanoseconds to milliseconds time-resolved spectra and kinetics of the intermediate states and species of bovine and carp fish rhodopsin visual pigments, which also contained ~5% cone pigments. The nanoseconds to milliseconds range cover all the major intermediates in the visual phototransduction process except the formation of bathorhodopsin intermediate which occurs at the femtosecond time scale. The dynamics of these visual pigment intermediates are initiated by excitation with a 532 nm nanosecond laser pulse. The recorded differences between bovine and carp rhodopsin time-resolved spectra of the formation and decay kinetics of their intermediates are presented and discussed. The data show that the carp samples batho intermediate decays faster, nearly by a factor of three, compared to the bovine samples. The formation and decay spectra and kinetics of rhodopsin outer segments and extracted rhodopsin inserted in buffer solution were found to be identical, with very small differences between them in the decay lifetimes of bathorhodopsin and formation of lumirhodopsin.

Prevalence of color vision deficiency and it’s relation with ABO blood group among medical students

Background: Normal human beings can appreciate color in all three of it’s attributes; Hue, intensity, and saturation. Human beings can perceive three primary colors, that is, red, green, and blue. Any defect in appreciation of colors is known as color vision deficiency (CVD). Complete inability to appreciate color is known as color blindness. The genes for red and green cone pigments are found in the q arm of the X chromosome. Hence, red and green CVD are inherited as X-linked recessive diseases. Blue CVD is autosomaly inherited. Blood groups are genetically inherited as well. Although it is a known fact that there is a preponderance of genetic diseases in subjects belonging to a specific blood group, no such study was available in Western Odisha as per our knowledge. This becomes more relevant as consanguineous marriages are a serious social problem in that part of Odisha. Aim and Objectives: Therefore, the study was undertaken to find out the prevalence of CVD among Medical students in Western Odisha and to find out it’s relation, if any, with ABO blood groups, as these two entities are genetically inherited. Record should be kept for future use, especially for counseling at the time of marriage and if necessary, to choose a subject for further study, where color detection does not play an important part. Materials and Methods: Prior permission was taken from the Institutional Ethics Committee to carry out this study. The study was carried out among the 1st–4th year medical students of V.S.S. Medical College and Hospital (n = 690) from August 2022 to October 2022. Ishihara test plates for color vision and agglutination method for ABO blood grouping were employed to get the prevalence of CVD and to get the percentage of subjects belonging to each blood group among those having CVD. The observations were noted carefully. Results: After careful screening, it was seen that 647 (93.76%) were normal, 42 (6.08%) had CVD, and 1 (0.14%) subject was completely color blind. All those affected were males. Distribution of blood group in CVD subjects showed 7 (16.27%) numbers belonging to blood group A, 22 (51.16%) belonging to blood group B, 1 (2.3%) belonging to blood group AB, and 12 (27.9%) belonging to blood group O. The only color blind student belonged to blood group B (2.3%). Conclusion: As medical students have to deal with colors throughout their career, the study helps the subjects to be aware of their condition early in their professional life. It also tried to find out the percentage distribution into ABO blood groups. A larger population should be included to get more accurate results.

Contribution of opsins and chromophores to cone pigment variation across populations of Lake Victoria cichlids.

Adaptation to heterogeneous sensory environments has been implicated as a key parameter in speciation. Cichlid fish are a textbook example of divergent visual adaptation, mediated by variation in the sequences and expression levels of cone opsin genes (encoding the protein component of visual pigments). In some vertebrates including fish, visual sensitivity is also tuned by the ratio of vitamin A1/A2–derived chromophores (i.e., the light‐sensitive component of the visual pigment bound to the opsin protein), where higher proportions of A2 cause a more red‐shifted wavelength absorbance. This study explores the variation in chromophore ratios across multiple cichlid populations in Lake Victoria, using as a proxy the expression of the gene Cyp27c1, which has been shown to regulate the conversion of vitamin A1 into vitamin A2 in several vertebrates. This study focuses on sympatric Pundamilia cichlids, where species with blue or red male coloration co‐occur at multiple islands but occupy different depths and consequently different visual habitats. In the red species, we found higher cyp27c1 expression in populations from turbid waters than from clear waters, but there was no such pattern in the blue species. Across populations, differences between the sympatric species in cyp27c1 expression had a consistent relationship with species differences in opsin expression patterns, but the red/blue identity reversed between clear and turbid waters. To assess the contribution of heritable vs. environmental causes of variation, we tested whether light manipulations induce a change in cyp27c1 expression in the laboratory. We found that cyp27c1 expression was not influenced by experimental light conditions, suggesting that the observed variation in the wild is due to genetic differences. Nonetheless, compared to other cichlid species, cyp27c1 is expressed at very low levels in Pundamilia, suggesting that it may not be relevant for visual adaptation in this species. Conclusively, establishing the biological importance of this variation requires testing of actual A1/A2 ratios in the eye, as well as its consequences for visual performance.

Color representations of normals and congenital red–green color deficiencies: Estimation of individual results based on color vision model

AbstractDifference scaling experiment was conducted to investigate the mental color representation of congenital color‐deficient observers and observers with normal color vision. Two decks of cards, high and medium chroma, each containing 10 Munsell chips, were prepared. A total of 45 pairs of hues were prepared for each of the decks. Ten protans, 10 deutans, and 10 people with normal color vision participated as observers, with each asked to rate the perceptual distance between two colors on a given color card. The results were analyzed using the multidimensional scaling method. All observers with normal color vision showed a circular shape close to the Munsell hue circle, while the majority of color‐deficient observers showed a concave shape bending at Y and PB. To indicate the degree of distortion from a circle, the distortion index was proposed to quantitatively evaluate intergroup and individual differences. To investigate the underlying mechanism of intergroup differences as well as individual differences in color representations of observers with normal color vision and color‐deficient observers, we proposed a model that considers various levels of human color vision mechanism from the cone pigment absorption, the luminance and opponent‐color coding level, and nonlinear transformation to difference‐scaling judgment. The circular shape for observers with normal color vision and some color‐deficient observers, as well as concave shapes for most color‐deficient observers were estimated. The correlation coefficient between the estimation and experiment‐based difference ranged from r = 0.64 to r = 0.94 with the grand average of r = 0.82, with p‐values less than 0.001 for all observers, suggesting that the concept of proposed model is appropriate.

Phototransduction in Anuran Green Rods: Origins of Extra-Sensitivity.

Green rods (GRs) represent a unique type of photoreceptor to be found in the retinas of anuran amphibians. These cells harbor a cone-specific blue-sensitive visual pigment but exhibit morphology of the outer segment typical for classic red rods (RRs), which makes them a perspective model object for studying cone–rod transmutation. In the present study, we performed detailed electrophysiological examination of the light sensitivity, response kinetics and parameters of discrete and continuous dark noise in GRs of the two anuran species: cane toad and marsh frog. Our results confirm that anuran GRs are highly specialized nocturnal vision receptors. Moreover, their rate of phototransduction quenching appeared to be about two-times slower than in RRs, which makes them even more efficient single photon detectors. The operating intensity ranges for two rod types widely overlap supposedly allowing amphibians to discriminate colors in the scotopic region. Unexpectedly for typical cone pigments but in line with some previous reports, the spontaneous isomerization rate of the GR visual pigment was found to be the same as for rhodopsin of RRs. Thus, our results expand the knowledge on anuran GRs and show that these are even more specialized single photon catchers than RRs, which allows us to assign them a status of “super-rods”.

Evolutionary adaptation of visual pigments in geckos for their photic environment.

Vertebrates generally have a single type of rod for scotopic vision and multiple types of cones for photopic vision. Noteworthily, nocturnal geckos transmuted ancestral photoreceptor cells into rods containing not rhodopsin but cone pigments, and, subsequently, diurnal geckos retransmuted these rods into cones containing cone pigments. High sensitivity of scotopic vision is underlain by the rod’s low background noise, which originated from a much lower spontaneous activation rate of rhodopsin than of cone pigments. Here, we revealed that nocturnal gecko cone pigments decreased their spontaneous activation rates to mimic rhodopsin, whereas diurnal gecko cone pigments recovered high rates similar to those of typical cone pigments. We also identified amino acid residues responsible for the alterations of the spontaneous activation rates. Therefore, we concluded that the switch between diurnality and nocturnality in geckos required not only morphological transmutation of photoreceptors but also adjustment of the spontaneous activation rates of visual pigments.

English

BACKGROUND Colour vision is a function of three types of cone pigments present in the retina. Colour vision deficiency is an important disorder of vision that may pose a handicap to the performance of an affected individual. The prevalence of colour blindness varies in different geographical areas. The identification and estimation of the prevalence of colour vision deficiency in school-going children will help to educate and guide the caregivers to help the children in selecting their profession. This study was done to estimate the prevalence, sex distribution, and types of colour vision deficiency among school-going children of 5 to 15 years. METHODS A cross-sectional observational study was done among 500 students to evaluate the colour vision during the period from 1st January 2018 to 30th June 2019 at the Regional Institute of Ophthalmology, Kolkata. Ishihara’s pseudo isochromatic colour vision chart 38th edition was used to assess the school children for colour vision status. The children who were found to be colour blind were further classified into degree and types of colour vision deficiency. RESULTS A total of 500 students (250 male & 250 female) of surrounding schools, in the age group of 5 years to 15 years, were screened. 480 students (96 %) had normal colour vision while 20 (4 %) students were found to have defective colour vision. Prevalence (4 %) for colour blindness was found to be higher in males (3.6 %) than females (0.4 %). It was observed that out of 20 (4 %) colour-blind subjects 3.6 % were protanopes and 0.4 % were deuteranopes. CONCLUSIONS The present study shows the prevalence of colour blindness found to be quite low (4 %) and more common in males (3.6 %) in comparison to females (0.4 %). Protanomaly (3.6 %) was more common than deuteranomaly (0.4 %). KEYWORDS Colour Blindness, Protanomaly, Deuteranomaly, School Children

COLOUR VISION DEFICIENCY AMONG MEDICAL STUDENTS OF CHITWAN

Background: Colour vision is the function of three types of cone pigments present in the retina. Though colours are used for important signs in the medical profession, screening for colour blindness is not paid attention at beginning of a medical career. This study was aimed to assess the presence of colour blindness, if any among the medical students of Chitwan Medical College. Methods: A total of 220 medical students including 127 females and 93 males aged between 16-24 years were examined for colour blindness. Students were shown the complete range of Ishihara’s plates under daylight conditions. The chart was held 75 cm away from the subjects. The types of colour vision deficiency were differentiated with help of key provided with the chart. The data was analyzed by using Statistical Package for the Social Sciences version 20. Results: Among 220 participants, 5(2.27%) were found to have colour vision deficiency. Out of 5colourdeficient students, 4of them were red-green colour vision deficient and 1 of them couldn’t appreciate colour (total colour blindness) within the speculated time for the test. Female partici­pants didn’t have colour vision deficiency while colour vision deficient students were unaware of their colour vision status. Conclusions: Colour vision deficiency affects male than female. Medical students should be screened for colour vision deficiency and made aware of their limitation so that they can take special care in their future clinical practice.