Retinoic acid signaling regulates spatiotemporal specification of human green and red cones.

Trichromacy is unique to primates among placental mammals, enabled by blue (short/S), green (medium/M), and red (long/L) cones. In humans, great apes, and Old World monkeys, cones make a poorly understood choice between M and L cone subtype fates. To determine mechanisms specifying M and L cones, we developed an approach to visualize expression of the highly similar M- and L-opsin mRNAs. M-opsin was observed before L-opsin expression during early human eye development, suggesting that M cones are generated before L cones. In adult human tissue, the early-developing central retina contained a mix of M and L cones compared to the late-developing peripheral region, which contained a high proportion of L cones. Retinoic acid (RA)-synthesizing enzymes are highly expressed early in retinal development. High RA signaling early was sufficient to promote M cone fate and suppress L cone fate in retinal organoids. Across a human population sample, natural variation in the ratios of M and L cone subtypes was associated with a noncoding polymorphism in the NR2F2 gene, a mediator of RA signaling. Our data suggest that RA promotes M cone fate early in development to generate the pattern of M and L cones across the human retina.

S-cone circuits in the primate retina for non-image-forming vision.

Intrinsically photosensitive retinal ganglion cells (ipRGCs) respond directly to light by virtue of containing melanopsin which peaks at about 483 nm. However, in primates, ipRGCs also receive color opponent inputs from short-wavelength-sensitive (S) cone circuits that are well-suited to encode circadian changes in the color of the sky that accompany the rising and setting sun. Here, we review the retinal circuits that endow primate ipRGCs with the cone-opponency capable of encoding the color of the sky and contributing to the wide-ranging effects of short-wavelength light on ipRGC-mediated non-image-forming visual function in humans.

From cones to color vision: a neurobiological model that explains the unique hues.

The irreducible unique hues-red, green, blue, and yellow-remain one of the great mysteries of vision science. Attempts to create a physiologically parsimonious model that can predict the spectral locations of the unique hues all rely on at least one post hoc adjustment to produce appropriate loci for unique green and unique red, and struggle to explain the non-linearity of the Blue/Yellow system. We propose a neurobiological color vision model that overcomes these challenges by using physiological cone ratios, cone-opponent normalization to equal-energy white, and a simple adaptation mechanism to produce color-opponent mechanisms that accurately predict the spectral locations and variability of the unique hues.

Potential value of color vision aids for varying degrees of color vision deficiency.

Red-green color vision deficiency (CVD) is the most common single locus genetic disorder in humans, affecting approximately 8% of males and 0.4% of females [G. H. M. Waaler, Acta Ophthalmol.5, 309 (2009)10.1111/j.1755-3768.1927.tb01016.x]; however, only about 1/4 of CVD individuals are dichromats who rely on only two cone types for color vision. The remaining 3/4 are anomalous trichromats whose CVD is milder, being based on three cone types, and who still perform remarkably well on many color-based tasks. To illustrate this, we have developed an algorithm that computes the relative loss of color discrimination in red-green CVD individuals with varying degrees of deficiency and accurately simulates their color experience for color normal observers. The resulting simulation illustrates the large gap in color discrimination between dichromats and even the most severe anomalous trichromats, showing that, relative to dichromats, the majority of anomalous trichromats can function without aids for color vision deficiency.

Tritan color vision deficiency may be associated with an OPN1SW splicing defect and haploinsufficiency.

Here we present evidence implicating disrupted RNA splicing as a potential cause of inherited tritan color vision. Initially we tested 51 subjects for color vision deficiencies. One made significant tritan errors; the others were classified as normal trichromats. The putative tritan subject was the only one of the 51 subjects found to be heterozygous for an OPN1SW gene mutation that disrupts RNA splicing in an in vitro assay. In order to gather further support for the role of the splicing mutation in tritan color vision, the putative tritan subject's mother and sister were examined. They also made tritan errors and had the same OPN1SW gene mutation.

Effect of cone spectral topography on chromatic detection sensitivity.

The spatial and spectral topography of the cone mosaic set the limits for detection and discrimination of chromatic sinewave gratings. Here, we sought to compare the spatial characteristics of mechanisms mediating hue perception against those mediating chromatic detection in individuals with known spectral topography and with optical aberrations removed with adaptive optics. Chromatic detection sensitivity in general exceeded previous measurements and decreased monotonically for increasingly skewed cone spectral compositions. The spatial grain of hue perception was significantly coarser than chromatic detection, consistent with separate neural mechanisms for color vision operating at different spatial scales.

Broad thorny ganglion cells: a candidate for visual pursuit error signaling in the primate retina.

Functional analyses exist only for a few of the morphologically described primate ganglion cell types, and their correlates in other mammalian species remain elusive. Here, we recorded light responses of broad thorny cells in the whole-mounted macaque retina. They showed ON-OFF-center light responses that were strongly suppressed by stimulation of the receptive field surround. Spike responses were delayed compared with parasol ganglion cells and other ON-OFF cells, including recursive bistratified ganglion cells and A1 amacrine cells. The receptive field structure was shaped by direct excitatory synaptic input and strong presynaptic and postsynaptic inhibition in both ON and OFF pathways. The cells responded strongly to dark or bright stimuli moving either in or out of the receptive field, independent of the direction of motion. However, they did not show a maintained spike response either to a uniform background or to a drifting plaid pattern. These properties could be ideally suited for guiding movements involved in visual pursuit. The functional characteristics reported here permit the first direct cross-species comparison of putative homologous ganglion cell types. Based on morphological similarities, broad thorny ganglion cells have been proposed to be homologs of rabbit local edge detector ganglion cells, but we now show that the two cells have quite distinct physiological properties. Thus, our data argue against broad thorny cells as the homologs of local edge detector cells.

ASSESSING PHOTORECEPTOR STRUCTURE ASSOCIATED WITH ELLIPSOID ZONE DISRUPTIONS VISUALIZED WITH OPTICAL COHERENCE TOMOGRAPHY.

PURPOSE: To compare images of photoreceptor layer disruptions obtained with optical coherence tomography (OCT) and adaptive optics scanning light ophthalmoscopy (AOSLO) in a variety of pathologic states. METHODS: Five subjects with photoreceptor ellipsoid zone disruption as per OCT and clinical diagnoses of closed-globe blunt ocular trauma (n = 2), macular telangiectasia type 2 (n = 1), blue-cone monochromacy (n = 1), or cone-rod dystrophy (n = 1) were included. Images were acquired within and around photoreceptor lesions using spectral domain OCT, confocal AOSLO, and split-detector AOSLO. RESULTS: There were substantial differences in the extent and appearance of the photoreceptor mosaic as revealed by confocal AOSLO, split-detector AOSLO, and spectral domain OCT en face view of the ellipsoid zone. CONCLUSION: Clinically available spectral domain OCT, viewed en face or as B-scan, may lead to misinterpretation of photoreceptor anatomy in a variety of diseases and injuries. This was demonstrated using split-detector AOSLO to reveal substantial populations of photoreceptors in areas of no, low, or ambiguous ellipsoid zone reflectivity with en face OCT and confocal AOSLO. Although it is unclear if these photoreceptors are functional, their presence offers hope for therapeutic strategies aimed at preserving or restoring photoreceptor function.

Circuitry to explain how the relative number of L and M cones shapes color experience.

The wavelength of light that appears unique yellow is surprisingly consistent across people even though the ratio of middle (M) to long (L) wavelength sensitive cones is strikingly variable. This observation has been explained by normalization to the mean spectral distribution of our shared environment. Our purpose was to reconcile the nearly perfect alignment of everyone's unique yellow through a normalization process with the striking variability in unique green, which varies by as much as 60 nm between individuals. The spectral location of unique green was measured in a group of volunteers whose cone ratios were estimated with a technique that combined genetics and flicker photometric electroretinograms. In contrast to unique yellow, unique green was highly dependent upon relative cone numerosity. We hypothesized that the difference in neural architecture of the blue-yellow and red-green opponent systems in the presence of a normalization process creates the surprising dependence of unique green on cone ratio. We then compared the predictions of different theories of color vision processing that incorporate L and M cone ratio and a normalization process. The results of this analysis reveal that-contrary to prevailing notions--postretinal contributions may not be required to explain the phenomena of unique hues.

Distinctive receptive field and physiological properties of a wide-field amacrine cell in the macaque monkey retina.

At early stages of visual processing, receptive fields are typically described as subtending local regions of space and thus performing computations on a narrow spatial scale. Nevertheless, stimulation well outside of the classical receptive field can exert clear and significant effects on visual processing. Given the distances over which they occur, the retinal mechanisms responsible for these long-range effects would certainly require signal propagation via active membrane properties. Here the physiology of a wide-field amacrine cell-the wiry cell-in macaque monkey retina is explored, revealing receptive fields that represent a striking departure from the classic structure. A single wiry cell integrates signals over wide regions of retina, 5-10 times larger than the classic receptive fields of most retinal ganglion cells. Wiry cells integrate signals over space much more effectively than predicted from passive signal propagation, and spatial integration is strongly attenuated during blockade of NMDA spikes but integration is insensitive to blockade of NaV channels with TTX. Thus these cells appear well suited for contributing to the long-range interactions of visual signals that characterize many aspects of visual perception.

Rod- and cone-driven responses in mice expressing human L-cone pigment.

The mouse is commonly used for studying retinal processing, primarily because it is amenable to genetic manipulation. To accurately study photoreceptor driven signals in the healthy and diseased retina, it is of great importance to isolate the responses of single photoreceptor types. This is not easily achieved in mice because of the strong overlap of rod and M-cone absorption spectra (i.e., maxima at 498 and 508 nm, respectively). With a newly developed mouse model (Opn1lw(LIAIS)) expressing a variant of the human L-cone pigment (561 nm) instead of the mouse M-opsin, the absorption spectra are substantially separated, allowing retinal physiology to be studied using silent substitution stimuli. Unlike conventional chromatic isolation methods, this spectral compensation approach can isolate single photoreceptor subtypes without changing the retinal adaptation. We measured flicker electroretinograms in these mutants under ketamine-xylazine sedation with double silent substitution (silent S-cone and either rod or M/L-cones) and obtained robust responses for both rods and (L-)cones. Small signals were yielded in wild-type mice, whereas heterozygotes exhibited responses that were generally intermediate to both. Fundamental response amplitudes and phase behaviors (as a function of temporal frequency) in all genotypes were largely similar. Surprisingly, isolated (L-)cone and rod response properties in the mutant strain were alike. Thus the LIAIS mouse warrants a more comprehensive in vivo assessment of photoreceptor subtype-specific physiology, because it overcomes the hindrance of overlapping spectral sensitivities present in the normal mouse.

Gene therapy for red-green colour blindness in adult primates.

Red-green colour blindness, which results from the absence of either the long- (L) or the middle- (M) wavelength-sensitive visual photopigments, is the most common single locus genetic disorder. Here we explore the possibility of curing colour blindness using gene therapy in experiments on adult monkeys that had been colour blind since birth. A third type of cone pigment was added to dichromatic retinas, providing the receptoral basis for trichromatic colour vision. This opened a new avenue to explore the requirements for establishing the neural circuits for a new dimension of colour sensation. Classic visual deprivation experiments have led to the expectation that neural connections established during development would not appropriately process an input that was not present from birth. Therefore, it was believed that the treatment of congenital vision disorders would be ineffective unless administered to the very young. However, here we show that the addition of a third opsin in adult red-green colour-deficient primates was sufficient to produce trichromatic colour vision behaviour. Thus, trichromacy can arise from a single addition of a third cone class and it does not require an early developmental process. This provides a positive outlook for the potential of gene therapy to cure adult vision disorders.

S-opsin knockout mice with the endogenous M-opsin gene replaced by an L-opsin variant.

Specific variants of human long-wavelength (L) and middle-wavelength (M) cone opsin genes have recently been associated with a variety of vision disorders caused by cone malfunction, including red-green color vision deficiency, blue cone monochromacy, myopia, and cone dystrophy. Strikingly, unlike disease-causing mutations in rhodopsin, most of the cone opsin alleles that are associated with vision disorders do not have deleterious point mutations. Instead, specific combinations of normal polymorphisms that arose by genetic recombination between the genes encoding L and M opsins appear to cause disease. Knockout/knock-in mice promise to make it possible to study how these deleterious cone opsin variants affect the structure, function, and viability of the cone photoreceptors. Ideally, we would like to evaluate different variants that cause vision disorders in humans against a control pigment that is not associated with vision disorders, and each variant should be expressed as the sole photopigment in each mouse cone, as is the case in humans. To evaluate the feasibility of this approach, we created a line of mice to serve as the control in the analysis of disease-causing mutations by replacing exon 2 through 6 of the mouse M-opsin gene with the corresponding cDNA for a human L-opsin variant that is associated with normal vision. Experiments reported here establish that the resulting pigment, which differs from the endogenous mouse M opsin at 35 amino acid positions, functions normally in mouse cones. This pigment was evaluated in mice with and without coexpression of the mouse short wavelength (S) opsin. Here, the creation and validation of two lines of genetically engineered mice that can be used to study disease-causing variants of human L/M-opsins, in vivo, are described.

Comparison of the Richmond HRR 4th edition and Farnsworth-Munsell 100 Hue Test for quantitative assessment of tritan color deficiencies.

Drugs and environmental factors can induce tritan deficiencies. The Farnsworth-Munsell (FM) 100 Hue Test has become the gold standard in measuring these acquired defects. However, the test is time consuming, and color discrimination is confounded by concentration and patience. Here, we describe a test that compares six tritan plates from the HRR Pseudoisochromatic Plates 4th edition to 16 FM 100 Hue tritan caps. CIE Standard Illuminant C was reduced over five light intensities to simulate the effects of acquired losses in the S-cone pathway. Both tests showed quantitative differences in error rates with all light levels; thus they could serve equally well for assessing acquired deficiencies. However, compared to the FM 100, the HRR took subjects about 20-40 s per trial, making it more practical.

Specialized synaptic pathway for chromatic signals beneath S-cone photoreceptors is common to human, Old and New World primates.

The distribution of the soluble NSF-attachment protein receptor protein syntaxin-4 and the Na-K-Cl cotransporter (NKCC) were investigated in the outer plexiform layer of human retina using immunohistochemistry. Both proteins, which are proposed to be components of a gamma-aminobutyric acid mediated feed-forward circuit from horizontal cells directly to bipolar cells, were enriched beneath S-cones. The expression pattern of syntaxin-4 was further analyzed in baboon and marmoset to determine if the synaptic specialization is common to primates. Syntaxin-4 was enriched beneath S-cones in both species, which together with the human results indicates that this specialization may have evolved for the purpose of mediating unique color vision capacities that are exclusive to primates.

Neurobiological hypothesis of color appearance and hue perception.

De Valois and De Valois [Vis. Res.33, 1053 (1993)] showed that to explain hue appearance, S-cone signals have to be combined with M versus L opponent signals in two different ways to produce red-green and yellow-blue axes, respectively. Recently, it has been shown that color appearance is normal for individuals with genetic mutations that block S-cone input to blue-ON ganglion cells. This is inconsistent with the De Valois hypothesis in which S-opponent konio-geniculate signals are combined with L-M signals at a third processing stage in cortex. Instead, here we show that color appearance, including individual differences never explained before, are predicted by a model in which S-cone signals are combined with L versus M signals in the outer retina.

Cone-isolating ON-OFF electroretinogram for studying chromatic pathways in the retina.

The electroretinogram (ERG) provides information about outer retina function in both clinical and research applications. ERG components elicited by light increments and decrements can be separated using a long-flash paradigm in which periods of light ON and OFF are alternated. Here, the ON-OFF ERG is combined with a silent substitution technique to elicit responses from individual cone photoreceptor classes by modulating the intensities of three color lights between the two periods. The results focus on the short wavelength (S) cone pathways since they are vulnerable to disease and because there are many unanswered questions about S-cone ON and OFF circuitry.

Diffusion Optics Technology (DOT): A Myopia Control Spectacle Lens Based on Contrast Theory.

Diffusion optics Technology (DOT) myopia control spectacle lenses are based on contrast theory. This innovative theory represents a radical departure from the classical concept of visual deprivation myopia. However, traditional theories have evolved, arriving at remarkably similar solutions for myopia control as the DOT lenses. Nonetheless, contrast theory still represents a departure from mainstream theories. Here, in an effort to resolve discrepancies, we review the science behind contrast theory and compare it to more conventional blur and defocus theories. Finally, we consider the implications of the different theories for the rational design of myopia control solutions.

Clarification on the understanding of contrast theory in relation to the article "ON and OFF receptive field processing in the presence of optical scattering": comment.

We are writing to address errors of misrepresentation in the article "ON and OFF receptive field processing in the presence of optical scattering" [Biomed. Opt. Express14, 2618 (2023)10.1364/BOE.489117]. In their investigation of predictions of "contrast theory" to explain the efficacy of diffusion optics technology (DOT), a myopia control lens design [Br. J. Ophthalmol.107, 1709 (2023)10.1136/bjo-2021-321005], Breher et al. incorrectly indicated that our contrast theory proposed that the association between cone opsin gene splicing defects and myopia was due to differential involvement in ON- and OFF-visual pathways. In addition, the Authors write that we have "hypothesized enhanced ON contrast sensitivity in myopes," but we predict the opposite.

Toward an Indoor Lighting Solution for Social Jet Lag.

There is growing interest in developing artificial lighting that stimulates intrinsically photosensitive retinal ganglion cells (ipRGCs) to entrain circadian rhythms to improve mood, sleep, and health. Efforts have focused on stimulating the intrinsic photopigment, melanopsin; however, specialized color vision circuits have been elucidated in the primate retina that transmit blue-yellow cone-opponent signals to ipRGCs. We designed a light that stimulates color-opponent inputs to ipRGCs by temporally alternating short- and long-wavelength components that strongly modulate short-wavelength sensitive (S) cones. Two-hour exposure to this S-cone modulating light produced an average circadian phase advance of 1 h and 20 min in 6 subjects (mean age = 30 years) compared to no phase advance for the subjects after exposure to a 500 lux white light equated for melanopsin effectiveness. These results are promising for developing artificial lighting that is highly effective in controlling circadian rhythms by invisibly modulating cone-opponent circuits.