Effects of color-enhancing glasses on color vision in congenital red-green color deficiencies.

As commercially available glasses for color vision deficiency (CVD) are classified as low risk, they are not subject to stringent marketing regulations. We investigate how EnChroma and VINO glasses affect performance on the Colour Assessment and Diagnosis (CAD) test in individuals with CVD. Data were obtained from 51 individuals with red-green CVD. Blood or saliva samples were collected to examine the structure of the OPN1LW/OPN1MW array. Individuals completed the CAD test twice without glasses and once with each pair of glasses. Although there was a statistically significant effect of both glasses, only that of VINO could be considered functionally meaningful.

Daily activity patterns influence retinal morphology, signatures of selection, and spectral tuning of opsin genes in colubrid snakes.

BACKGROUND: Morphological divergences of snake retinal structure point to complex evolutionary processes and adaptations. The Colubridae family has a remarkable variety of retinal structure that can range from all-cone and all-rod to duplex (cone/rod) retinas. To explore whether nocturnal versus diurnal activity is responsible for constraints on molecular evolution and plays a role in visual opsin spectral tuning of colubrids, we carried out molecular evolution analyses of the visual opsin genes LWS, RH1, and SWS1 from 17 species and performed morphological analyses. RESULTS: Phylogenetic reconstructions of the RH1 and LWS recovered major clades characterized by primarily diurnal or primarily nocturnal activity patterns, in contrast with the topology for SWS1, which is very similar to the species tree. We found stronger signals of purifying selection along diurnal and nocturnal lineages for RH1 and SWS1, respectively. A blue-shift of the RH1 spectral peak is associated with diurnal habits. Spectral tuning of cone opsins did not differ among diurnal and nocturnal species. Retinas of nocturnal colubrids had many rows of photoreceptor nuclei, with large numbers of rods, labeled by wheat germ agglutinin (WGA), and two types of cones: large cones sensitive to long/medium wavelengths (L/M) and small cones sensitive to ultra-violet/violet wavelengths (UV/VS). In contrast, retinas of diurnal species had only one row of photoreceptor nuclei, with four types of cones: large and double L/M cones, small UV/VS cones, and a second group of small cones, labeled by WGA. CONCLUSIONS: For LWS gene, selection tests did not confirm different constraints related to activity pattern. For SWS1, stronger purifying selection in nocturnal lineages indicates divergent evolutionary pressures related to the activity pattern, and the importance of the short wavelength sensitivity at low light condition. Activity pattern has a clear influence on the signatures of selection and spectral tuning of RH1, with stronger purifying selection in diurnal lineages, which indicates selective pressure to preserve rhodopsin structure and function in pure-cone retinas. We suggest that the presence of four cone types in primarily diurnal colubrids might be related to the gain of color discrimination capacity.

Evolution of the circuitry for conscious color vision in primates.

There are many ganglion cell types and subtypes in our retina that carry color information. These have appeared at different times over the history of the evolution of the vertebrate visual system. They project to several different places in the brain and serve a variety of purposes allowing wavelength information to contribute to diverse visual functions. These include circadian photoentrainment, regulation of sleep and mood, guidance of orienting movements, detection and segmentation of objects. Predecessors to some of the circuits serving these purposes presumably arose before mammals evolved and different functions are represented by distinct ganglion cell types. However, while other animals use color information to elicit motor movements and regulate activity rhythms, as do humans, using phylogenetically ancient circuitry, the ability to appreciate color appearance may have been refined in ancestors to primates, mediated by a special set of ganglion cells that serve only that purpose. Understanding the circuitry for color vision has implications for the possibility of treating color blindness using gene therapy by recapitulating evolution. In addition, understanding how color is encoded, including how chromatic and achromatic percepts are separated is a step toward developing a complete picture of the diversity of ganglion cell types and their functions. Such knowledge could be useful in developing therapeutic strategies for blinding eye disorders that rely on stimulating elements in the retina, where more than 50 different neuron types are organized into circuits that transform signals from photoreceptors into specialized detectors many of which are not directly involved in conscious vision.

A study of unusual Rayleigh matches in deutan deficiency.

Rayleigh match data were modeled with the aim of explaining the locations of match midpoints and matching ranges, both in normal trichromats and in subjects with congenital color deficiency. Model parameters included the wavelength of peak sensitivity of cone photopigments, the effective photopigment optical density, and the noise amplitude in the red-green color channel. In order to avoid the suprathreshold, perceptual effects of extreme L:M cone ratios on color vision, selective post-receptoral amplification of cone signals is needed. The associated noise is also amplified and this causes corresponding changes in red-green threshold sensitivity. We propose that the noise amplitude and hence the size of the matching range in normal trichromats relates to the known inter-subject variation in the relative numbers of L and M cones. If this hypothesis can be shown to account for the extremes of the red-green matching range measured in normal trichromats, it is of interest to establish the extent to which it also predicts the unexpected, small matching ranges that are observed in some subjects with red-green color deficiency. A subset of subjects with deutan deficiency that exhibited less common Nagel matches were selected for genetic analysis of their cone pigment genes in order to confirm the type of deficiency, and to predict the corresponding peak wavelength separation (delta lambda(max)) of their two, long-wavelength cone pigments. The Rayleigh match model predicted accurately the midpoint and the range for the spectral differences specified by the genes. The prediction also required plausible selection of effective optical density of the cone pigments and noise. The noise needed varied, but the estimates were confined to lie within the limits established from the matching ranges measured in normal trichromats. The model predicts correctly the small matching ranges measured in some deuteranomalous subjects, principally accounted for by a low estimate of noise level in the red-green channel. The model also predicts the "normal" matches made by some subjects that rely on two hybrid genes and therefore exhibit red-green thresholds outside the normal range, typical of mild deuteranomaly.

Polymorphism of visual pigment genes in the muriqui (Primates, Atelidae).

Colour vision varies within the family Atelidae (Primates, Platyrrhini), which consists of four genera with the following cladistic relationship: {Alouatta[Ateles (Lagothrix and Brachyteles)]}. Spider monkeys (Ateles) and woolly monkeys (Lagothrix) are characteristic of platyrrhine monkeys in possessing a colour vision polymorphism. The polymorphism results from allelic variation of the single-locus middle-to-long wavelength (M/L) cone opsin gene on the X-chromosome. The presence in the population of alleles coding for different M/L photopigments results in a variety of colour vision phenotypes. Such a polymorphism is absent in howling monkeys (Alouatta), which, alone among platyrrhines, acquired uniform trichromatic vision similar to that of Old World monkeys, apes, and humans through opsin gene duplication. Dietary and morphological similarities between howling monkeys and muriquis (Brachyteles) raise the possibility that the two genera share a similar form of colour vision, uniform trichromacy. Yet parsimony predicts that the colour vision of Brachyteles will resemble the polymorphism present in Lagothrix and Ateles. Here we test this assumption. We obtained DNA from the blood or faeces of 18 muriquis and sequenced exons 3 and 5 of the M/L opsin gene. Our results affirm the existence of a single M/L cone opsin gene in the genus Brachyteles. We detected three alleles with predicted lambdamax values of 530, 550, and 562 nm. Two females were heterozygous and are thus predicted to have different types of M/L cone pigment. We discuss the implication of this result towards understanding the evolutionary ecology of trichromatic vision.

The uncommon retina of the common house mouse.

Unlike most mammals, most cones in house mouse retina express two opsins, one sensitive to UV-light, and another sensitive to middle-wavelengths. Is the mouse unique, having a single cone type that normally expresses two opsins? Or is the mouse a typical mammal having two cone types, but a species wide mutation results in co-expression of two opsins?

Photopigment basis for dichromatic color vision in the horse.

Horses, like other ungulates, are active in the day, at dusk, dawn, and night; and, they have eyes designed to have both high sensitivity for vision in dim light and good visual acuity under higher light levels (Walls, 1942). Typically, daytime activity is associated with the presence of multiple cone classes and color-vision capacity (Jacobs, 1993). Previous studies in other ungulates, such as pigs, goats, cows, sheep and deer, have shown that they have two spectrally different cone types, and hence, at least the photopigment basis for dichromatic color vision (Neitz & Jacobs, 1989; Jacobs, Deegan II, Neitz, Murphy, Miller, & Marchinton, 1994; Jacobs, Deegan II, & Neitz, 1998). Here, electroretinogram flicker photometry was used to measure the spectral sensitivities of the cones in the domestic horse (Equus caballus). Two distinct spectral mechanisms were identified and are consistent with the presence of a short-wavelength-sensitive (S) and a middle-to-long-wavelength-sensitive (M/L) cone. The spectral sensitivity of the S cone was estimated to have a peak of 428 nm, while the M/L cone had a peak of 539 nm. These two cone types would provide the basis for dichromatic color vision consistent with recent results from behavioral testing of horses (Macuda & Timney, 1999; Macuda & Timney, 2000; Timney & Macuda, 2001). The spectral peak of the M/L cone photopigment measured here, in vivo, is similar to that obtained when the gene was sequenced, cloned, and expressed in vitro (Yokoyama & Radlwimmer, 1999). Of the ungulates that have been studied to date, all have the photopigment basis for dichromatic color vision; however, they differ considerably from one another in the spectral tuning of their cone pigments. These differences may represent adaptations to the different visual requirements of different species.

Molecular genetics of color vision and color vision defects.

Color is an extremely important component of the information that we gather with our eyes. Most of us use color so automatically that we fail to appreciate how important it is in our daily activities. It serves as a nonlinguistic code that gives us instant information about the world around us. From observing color, for example, we can find the bee sting on an infant's arm even before it begins to swell by looking for the little spot where the infant's skin is red. We know when fruit is ripe; the ripe banana is yellow not green. We know when meat is cooked because it is no longer red. When watching a football game, we can instantly keep track of the players on opposing teams from the colors of their uniforms. Using color, we know from a distance which car is ours in the parking lot--it is the blue one--and whether we will need to stop at the distant traffic light, even at night, when we cannot see the relative positions of red and green lights.

Flicker-photometric electroretinogram estimates of L:M cone photoreceptor ratio in men with photopigment spectra derived from genetics.

Relative proportions of long-wavelength-sensitive (L) to middle-wavelength-sensitive (M) cones were estimated by use of the flicker-photometric electroretinogram (ERG). It has been demonstrated that a major source of error in estimates of cone proportions from spectral luminosity functions is the known variation in the lambda(max) of the photopigments [Vision Res. 38, 1961 (1998)]. To correct for these errors, estimates of cone proportions were derived by use of individualized L-cone spectral sensitivity curves deduced from photopigment gene sequences from each subject. For some individuals this correction made a large difference in the estimated cone proportions compared with the value obtained when a fixed standard L cone was assumed. The largest discrepancy occurred in a man estimated to have 62% L cones (L:M ratio 1.6:1) when a standard L pigment was assumed but a value of 80% L cones (L:M ratio 4:1) when his individualized L-cone spectrum was used. From repeated measurements made with the ERG, it was determined that individual estimates of the relative L-to-M cone contributions, expressed as %L cones, are usually reliable within approximately 2%. The average L:M ratio for 15 male subjects was estimated at 2:1 (67% L cones). Previously, a large range of individual variability was reported for L:M ratios obtained from photometry. An unresolved issue concerns how much of the range might be attributed to error. Here efforts have been taken to markedly reduce measurement error. Nonetheless, a large range of individual differences persists. Estimated L:M ratios for individuals ranged from 0.6:1 to 12:1 (40% L to 92% L).

Cone pigment gene expression in individual photoreceptors and the chromatic topography of the retina.

Human trichromatic vision is based on three classes of cones: L, M, and S (long-, middle-, and short-wavelength sensitive, respectively). Individuals can have more than one M and/or more than one L pigment gene on the X chromosome along with an S pigment gene on chromosome 7. In some people the X-linked pigment gene array can include polymorphic variants that encode multiple, spectrally distinct cone photopigment subtypes. A single-cell, polymerase chain reaction approach was used to examine visual pigment gene expression in individual human cone cells and identify them as L or M. The ratio of L:M pigment gene expression was assayed in homogenized retinal tissues taken from the same eyes. Results indicate that there is a close correspondence between the cone ratio determined from counting single cells and the L:M pigment mRNA ratio estimated from homogenized pieces of retina. The results also show that the different pigment genes in one array are often expressed at very different levels, giving rise to unequal numbers of L and M cones. Expression of only one photopigment gene was detected in each cone cell. However, individual males can have more than the classically described three spectrally distinct cone types in their retinas.

Functional consequences of the relative numbers of L and M cones.

Direct imaging of the retina by adaptive optics allows assessment of the relative number of long-wavelength-sensitive (L) and middle-wavelength-sensitive (M) cones in living human eyes. We examine the functional consequences of variation in the relative numbers of L and M cones (L/M cone ratio) for two observers whose ratios were measured by direct imaging. The L/M cone ratio for the two observers varied considerably, taking on values of 1.15 and 3.79. Two sets of functional data were collected: spectral sensitivity measured with the flicker electroretinogram (ERG) and the wavelength of unique yellow. A genetic analysis was used to determine L and M cone spectra appropriate for each observer. Rayleigh matches confirmed the use of these spectra. We determined the relative strength of L and M cone contributions to ERG spectral sensitivity by fitting the data with a weighted sum of L and M cone spectra. The relative strengths so determined (1.06 and 3.38) were close to the cone ratios established by direct imaging. Thus variation in L/M cone ratio is preserved at the sites tapped by the flicker ERG. The wavelength of unique yellow varied only slightly between the two observers (576.8 and 574.7 nm). This small variation indicates that neural factors play an important role in stabilizing unique yellow against variation in the L/M cone ratio.

Vitronectin gene expression in the adult human retina.

PURPOSE: To determine whether vitronectin (Vn), a plasma protein and extracellular matrix molecule that is also a prominent constituent of drusen, is synthesized by cells in the adult human retina. METHODS: The distribution of Vn in the normal adult human retina was examined using antibodies to circulating plasma Vn and to the multimeric, heparin-binding form that is most prevalent in extravascular tissues. Evidence of Vn transcription by retinal cells was analyzed by in situ hybridization and also by reverse transcription of total RNA derived from dissociated human or mouse photoreceptors followed by amplification using polymerase chain reaction (RT-PCR). RESULTS: Cytoplasmic immunoreactivity for plasma Vn or multimeric Vn was detected in photoreceptors, in a subpopulation of neurons situated in the inner retina, and in vitreous hyalocytes. Extracellular labeling was limited primarily to Bruch's membrane and the retinal vasculature. At the transcriptional level, Vn mRNA was localized to both photoreceptors and ganglion cells by in situ hybridization. The in situ findings were corroborated by RT-PCR using total RNA from dissociated mouse or human photoreceptor cells. CONCLUSIONS: The results constitute the first evidence for Vn gene expression by adult neurons in the mammalian central nervous system. The identification of the photoreceptors as a cellular source of Vn suggests that these cells have the potential to make a biosynthetic contribution to the Vn that is found in drusen.

Trichromatic color vision with only two spectrally distinct photopigments.

Protanomaly is a common, X-linked abnormality of color vision. Like people with normal color vision, protanomalous observers are trichromatic, but their ability to discriminate colors in the red-green part of the spectrum is reduced because the photopigments that mediate discrimination in this range are abnormally similar. Whereas normal subjects have pigments whose wavelengths of peak sensitivity differ by about 30 nm, the peak wavelengths for protanomalous observers are thought to differ by only a few nanometers. We found, however, that although this difference occurred in some protanomalous subjects, others had pigments whose peak wavelengths were identical. Genetic and psychophysical results from the latter class indicated that limited red-green discrimination can be achieved with pigments that have the same peak wavelength sensitivity and that differ only in optical density. A single amino acid substitution was correlated with trichromacy in these subjects, suggesting that differences in pigment sequence may regulate the optical density of the cone.

Variations in cone populations for red-green color vision examined by analysis of mRNA.

In the central human retina, there are estimated to be nearly two L cone photoreceptors for each M cone. The extent to which this value varies across individuals is unclear and little is known about how the M:L cone ratio might change with retinal location. To address these questions, the ratio of M:L cone pigment mRNA was examined at different locations. For patches of central retina, the average M:L ratio was about 2:3 which decreased to about 1:3 for patches 40 degrees eccentric. There were also large individual differences among the 23 eyes examined. The extremes differed in central M:L mRNA ratio by a factor of > 3. The measured differences in mRNA ratio are proposed to reflect differences in photoreceptor ratio. Such variations provide unique opportunities for understanding how the neural circuitry for color vision is affected by changes in cone ratio.

Spectra of human L cones.

Variations in the amino acid sequences of the human cone opsins give rise to spectrally variant subtypes of L and M cone pigments even in the population with normal color vision. In vitro mutagenesis studies have shown that a limited number of amino acid substitutions produce shifts in the wavelength sensitivity. Presented here are results comparing electrophysiological measurements of single human cones with the expressed cone pigment gene sequences from the same retina. In a sample of eight long-wavelength sensitive cone (L cone) spectra obtained from five donors the precise spectral sensitivities, measured in situ, of the two most commonly occurring spectral variants were determined. The peak sensitivity of the Lser180 cone was 563 nm while that of the Lala180 cone was 559 nm.

L-cone pigment genes expressed in normal colour vision.

To directly test the hypothesis that only two pigment genes are expressed from the X-chromosome array, we examined expressed M and L pigment gene sequences from > 100 male eye donors. In this sample, there were eight men who expressed high levels of more than one L pigment gene in addition to M pigment genes. The fact that these eyes expressed both L and M pigment genes at significant levels suggests they were from men with normal colour vision. We reject the hypothesis that only two pigment genes from one X-chromosome array can be expressed.

Expression of L cone pigment gene subtypes in females.

Spectral subtypes of L pigment are produced by a serine/alanine dimorphism at amino acid position 180. X-chromosomes that carry genes for the different subtypes occur with about equal frequency in normal men. Females have two X-chromosomes: thus, about 50% of women will inherit genes for both L pigment subtypes, although on different X-chromosomes. In these women, X-inactivation is expected to produce about equal numbers of LS180 and LA180 cones in addition to middle (M) and short (S) wavelength-sensitive cones to total four spectrally distinct cone types. Consistent with this expectation we found nearly equal expression of genes for two spectrally distinct subtypes of L pigment in five of nine female retinas examined.

M- and L-cones in early infancy: III. Comparison of genotypic and phenotypic markers of color vision in infants and adults.

Genetic analyses were performed on five male children (approximately 3 years), two suspect color-normals and three suspects for congenital color vision deficiencies. These classifications were based on visually-evoked potential (VEP) responses to M- and L-cone-isolating stimuli obtained in a previous study when each subject was either 4- or 8-weeks old. The present analyses were performed in a blind study to characterize the genotypes of these subjects. Four male adults with various color vision phenotypes were also tested as a control. DNA was isolated using a non-invasive technique followed by polymerase chain reaction (PCR) amplification and restriction enzyme analysis to examine the genomic DNA of each subject. The genetic analyses confirmed the VEP identification of two color defective infants, and were consistent with the diagnosis of two other infants as color normal. A third infant was predicted by VEP analysis to have a protan defect, but he did not have a gene array typically found in protan observers.

Recent evolution of uniform trichromacy in a New World monkey.

Until recently, New World primates were found to have a single M/L photopigment gene on the X-chromosome. This arrangement limits males to dichromatic, or monochromatic color vision. Only females who were heterozygous for the M/L gene were trichromatic. Recently, an exception has been discovered. Male howler monkeys appear to have more than one M/L pigment gene, and both genders are uniformly trichromatic. We characterized promoter regions corresponding to two M/L pigment genes in howlers. Comparison of DNA sequences with those of humans and three species of New World primate suggest a recent and independent acquisition of a second M/L gene locus in the howler.

Pigment gene expression in protan color vision defects.

We screened 150 male eye donors and identified four who did not have or express L pigment genes, consistent with each of them having a congenital protan color vision defect. One donor was identified as a protanope because he had and expressed a single X-chromosome photopigment gene that encoded an M pigment. Three were categorized as protanomalous because each expressed significant levels of genes specifying two spectrally different M pigments. The first gene in each of the protanomalous arrays was expressed the most and encoded an M pigment that differed in amino acid sequence from M pigments in color normal men.