The 19th Symposium of the International Colour Vision Society (ICVS) was held in Belém, Brazil in July 2007. This was the first time that the Society has met in Latin America. The Amazonian region was a unique location for this inauguration due both to its situation close to a vast, largely unexplored region of nature, and resources—an inspiring challenge for scientific study—and to the quality of the vision research developing in the region.

Neural models of retinal processing provide an important tool for analyzing retinal signals and their functional significance. However, it is here argued that in biological reality, retinal connectivity is unlikely to be as specific as ideal neural models might suggest. The retina is thought to provide functionally specific signals, but this specificity is unlikely to be anatomically complete. This is illustrated by examples of cone connectivity to macaque ganglion cells. For example, cells of the magnocellular pathway appear to avoid short-wavelength cone input, so that such input is negligible under normal conditions. However, there is anatomical, physiological, and psychophysical evidence that under special conditions, weak input may be revealed. Second, ideal models of how retinal information is centrally utilized have to take into account the biological reality of retinal signals. The stochastic nature of impulse trains modifies signal-to-noise ratio in unexpected ways. Also, non-linearities in cell responses make, for example, multiplexing of luminance and chromatic signals in the parvocellular pathway impracticable. The purpose of this analysis is to show than ideal neural models must confront an often more complex and nuanced physiological reality.

Electrophysiological and molecular genetic studies have shown that howler monkeys (Alouatta) are unique among all studied platyrrhines: they have the potential to display trichromatic color vision among males and females. This study examined the color discrimination abilities of four howler monkeys (Alouatta caraya) through a series of tasks involving a behavioral paradigm of discrimination learning. The animals were maintained and housed as a group in the Zoological Gardens of Brasília and were tested in their own home cages. Stimuli consisting of pairs of Munsell color chips were presented in random brightness values to assure that discriminations were based on color rather than brightness cues. All the animals (three males, one female) successfully discriminated all the stimulus pairs, including those that would be expected to be difficult for a dichromatic monkey. These results are consistent with the earlier predictions suggesting that howler monkeys are routinely trichromatic.

During their complex life history, anguilliform eels go through a major metamorphosis when developing from a fresh water yellow eel into a deep-sea silver eel. In addition to major changes in body morphology, the visual system also adapts from a fresh water teleost duplex retina with rods and cones, to a specialized deep-sea retina containing only rods. The history of the rods is well documented with an initial switch from a porphyropsin to a rhodopsin (P5232 to P5011) and then a total change in gene expression with the down regulation of a “freshwater” opsin and its concomitant replacement by the expression of a typical “deep-sea” opsin (P5011 to P4821). Yellow eels possess only two spectral classes of single cones, one sensitive in the green presumably expressing an RH2 opsin gene and the second sensitive in the blue expressing an SWS2 opsin gene. In immature glass eels, entering into rivers from the sea, the cones contain mixtures of rhodopsins and porphyropsins, whereas the fully freshwater yellow eels have cone pigments that are almost pure porphyropsins with peak sensitivities at about 540–545 nm and 435–440 nm, respectively. However, during the early stages of metamorphosis, the pigments switch to rhodopsins with the maximum sensitivity of the “green”-sensitive cone shifting to about 525 nm, somewhat paralleling, but preceding the change in rods. During metamorphosis, the cones are almost completely lost.

We have determined the sequence and genomic organization of the genes encoding the cone visual pigment of the platypus (Ornithorhynchus anatinus) and the echidna (Tachyglossus aculeatus), and inferred their spectral properties and evolutionary pathways. We prepared platypus and echidna retinal RNA and used primers of the middle-wave-sensitive (MWS), long-wave-sensitive (LWS), and short-wave sensitive (SWS1) pigments corresponding to coding sequences that are highly conserved among mammals; to PCR amplify the corresponding pigment sequences. Amplification from the retinal RNA revealed the expression of LWS pigment mRNA that is homologous in sequence and spectral properties to the primate LWS visual pigments. However, we were unable to amplify the mammalian SWS1 pigment from these two species, indicating this gene was lost prior to the echidna-platypus divergence (∼21 MYA). Subsequently, when the platypus genome sequence became available, we found an LWS pigment gene in a conserved genomic arrangement that resembles the primate pigment, but, surprisingly we found an adjacent (∼20 kb) SWS2 pigment gene within this conserved genomic arrangement. We obtained the same result after sequencing the echidna genes. The encoded SWS2 pigment is predicted to have a wavelength of maximal absorption of about 440 nm, and is paralogous to SWS pigments typically found in reptiles, birds, and fish but not in mammals. This study suggests the locus control region (LCR) has played an important role in the conservation of photo receptor gene arrays and the control of their spatial and temporal expression in the retina in all mammals. In conclusion, a duplication event of an ancestral cone visual pigment gene, followed by sequence divergence and selection gave rise to the LWS and SWS2 visual pigments. So far, the echidna and platypus are the only mammals that share the gene structure of the LWS-SWS2 pigment gene complex with reptiles, birds and fishes.

In support of the long-held idea that cone ratio is genetically determined by variation linked to the X-chromosome opsin gene locus, the present study identified nucleotide differences in DNA segments containing regulatory regions of the L and M opsin genes that are associated with significant differences in the relative number of L versus M cones. Specific haplotypes (combinations of genetic differences) were identified that correlated with high versus low L:M cone ratio. These findings are consistent with the biological principle that DNA sequence variations affect binding affinities for protein components of complexes that influence the relative probability that an L versus M opsin gene will be silenced during development, and in turn, produce variation in the proportion of L to M cones.

Delivery of foreign opsin genes to cone photoreceptors using recombinant adeno-associated virus (rAAV) is a potential tool for studying the basic mechanisms underlying cone based vision and for treating vision disorders. We used an in vivo retinal imaging system to monitor, over time, expression of virally-delivered genes targeted to cone photoreceptors in the Mongolian gerbil (Meriones unguiculatus). Gerbils have a well-developed photopic visual system, with 11–14% of their photoreceptors being cones. We used replication deficient serotype 5 rAAV to deliver a gene for green fluorescent protein (GFP). In an effort to direct expression of the gene specifically to either S or M cones, the transgene was under the control of either the human X-chromosome opsin gene regulatory elements, i.e., an enhancer termed the locus control region (LCR) and L promoter, or the human S-opsin promoter. Longitudinal fluorescence images reveal that gene expression is first detectable about 14 days post-injection, reaches a peak after about 3 months, and is observed more than a year post-injection if the initial viral concentration is sufficiently high. The regulatory elements are able to direct expression to a subpopulation of cones while excluding expression in rods and non-photoreceptor retinal cells. When the same viral constructs are used to deliver a human long-wavelength opsin gene to gerbil cones, stimulation of the introduced human photopigment with long-wavelength light produces robust cone responses.

Many attempts have been made over the years to distinguish human and primate L (long-wavelength sensitive) from M (middle-wavelength sensitive) cone photoreceptors using either immunohistochemistry or in situ hybridization. These attempts have been unsuccessful due to the very high degree of identity between the sequences of the L and M proteins and encoding mRNAs. The recent development of chemically modified oligonucleotide probes, referred to as locked nucleic acid (LNA) probes, has shown that they hybridize with much greater affinity and specificity to the target nucleic acid. This has greatly increased the potential for differentiating L from M cones by in situ hybridization. We have designed LNA oligonucleotide probes that are complementary to either the L or M coding sequences located in exon 5 of the Macaca nemestrina L and M pigment genes. We have shown that the LNA-M and LNA-L probes hybridize specifically to their respective target nucleic acid sequences in vitro. This result strongly suggests that these probes would be instrumental in rapidly distinguishing L from M cone in the entire retina, and in defining the cone mosaic during development and in adults.

To better understand the evolution of spatial and color vision, the number and spatial distributions of cones, rods, and optic nerve axon numbers were assessed in seven New World primates (Cebus apella, Saimiri ustius, Saguinus midas niger, Alouatta caraya, Aotus azarae, Calllithrix jacchus, and Callicebus moloch). The spatial distribution and number of rods and cones was determined from counts of retinal whole mounts. Optic axon number was determined from optic nerve sections by electron microscopy. These data were amassed with existing data on retinal cell number and distribution in Old World primates, and the scaling of relative densities and numbers with respect to retinal area, eye and brain sizes, and foveal specializations were evaluated. Regular scaling of all cell types was observed, with the exceptionally large, rod-enriched retina of the nocturnal owl monkey Aotus azarae, and the unusually high cone density of the fovea of the trichromatic howler monkey Alouatta caraya presenting interesting variations on this basic plan. Over all species, the lawful scaling of rods, cones, and retinal ganglion cell number is hypothesized to result from a conserved sequence of cell generation that defends retinal acuity and sensitivity over a large range of eye sizes.

The topographical distribution of relative sensitivity to red and green lights across the retina was assayed using a custom-made wide-field color multifocal electroretinogram apparatus. There were increases in the relative sensitivity to red compared to green light in the periphery that correlate with observed increases in the relative amount of long (L) compared to middle (M) wavelength sensitive opsin mRNA. These results provide electrophysiological evidence that there is a dramatic increase in the ratio of L to M cones in the far periphery of the human retina. The central to far peripheral homogeneity in cone proportions has implications for understanding the developmental mechanisms that determine the identity of a cone as L or M and for understanding the circuitry for color vision in the peripheral retina.

The turtle retina has been extensively used for the study of chromatic processing mechanisms. Color opponency has been previously investigated with trichromatic paradigms, but behavioral studies show that the turtle has an ultraviolet (UV) channel and a tetrachromatic visual system. Our laboratory has been working in the characterization of neuronal responses in the retina of vertebrates using stimuli in the UV-visible range of the electromagnetic spectrum. In the present investigation, we recorded color-opponent responses from turtle amacrine and ganglion cells to UV and visible stimuli and extended our previous results that UV color-opponency is present at the level of the inner nuclear layer. We recorded from 181 neurons, 36 of which were spectrally opponent. Among these, there were 10 amacrine (5%), and 26 ganglion cells (15%). Morphological identification of color-opponent neurons was possible for two ganglion cell classes (G17 and G22) and two amacrine cell classes (A22 and A23b). There was a variety of cell response types and a potential for complex processing of chromatic stimuli, with intensity- and wavelength-dependent response components. Ten types of color opponency were found in ganglion cells and by adding previous results from our laboratory, 12 types of opponent responses have been found. The majority of the ganglion cells were R+UVBG- and RG+UVB-color-opponents but there were other less frequent types of chromatic opponency. This study confirms the participation of a UV channel in the processing of color opponency in the turtle inner retina and shows that the turtle visual system has the retinal mechanisms to allow many possible chromatic combinations.

We investigated how the stimulation mode influences transient visual evoked potentials (tVEP) amplitude as a function of contrast of achromatic and isoluminant chromatic gratings. The chromatic stimulation probed only responses to the red-green axis. Visual stimuli were monocularly presented in a 5° diameter circle, achromatic and chromatic horizontal gratings, 1 Hz pattern reversal stimulation, and achromatic and chromatic gratings, 300 ms onset per 700 ms offset stimulation. For the achromatic pattern reversal stimulation, a double slope function describes how the P100 amplitude varied as a function of log contrast which had a limb at low-to-medium contrasts and another limb at high contrasts. For the achromatic onset/offset stimulation, C2 amplitude saturated at the highest contrast tested and a single straight line described how it changed along most of the contrast range. Both presentation modes for chromatic gratings resulted in amplitude versus log contrast relations which were well described by single straight lines along most of the contrast range. The results may be interpreted as if at 2 cpd, achromatic pattern reversal stimulation evoked the activity of at least two visual pathways with high and low contrast sensitivity, respectively, while achromatic onset/offset stimulation favored the activity of a pathway with high contrast sensitivity. The neural activity in the M pathway is the best candidate to be the high contrast mechanism detected with pattern reversal and pattern onset/offset VEPs. The activity of color opponent pathways such as the P and K pathways either combined or in isolation seems to be responsible for VEPs obtained with isoluminant chromatic gratings at both presentation modes. When the amplitudes of chromatic VEPs were plotted in the same contrast scale as used for achromatic VEPs, chromatic contrast thresholds had similar values to those of the achromatic mechanism with high contrast sensitivity.

Using double silent substitution, it is possible to generate L-cone and M-cone isolating electroretinograms (ERGs) on a CRT. A major limitation of the technique is that the depth of modulation of cone classes is limited by the restricted luminance of the phosphors and their spectral overlap. To address this problem we have ported the technique to a four-color LED Ganzfeld stimulus (Diagnosis ColorDome). This allows higher retinal illuminances, higher contrasts, and triple silent substitution. With careful control over the retinal area stimulated, we show that the same data can be recorded from both CRT and LED stimuli when luminance, size and cone contrast are kept constant. Importantly, the different temporal profiles of the two devices do not influence the ERG amplitude and phase plots. We present data over a much wider range of luminances (up to about 10000 trolands) and contrasts with the LED stimulator than previously reported with CRT screens. We conclude that the close resemblance between data obtained with an LED stimulator and with a CRT screen indicate that the differences have a purely physiological origin.

The purpose of this work is to investigate the use of different forms of visual evoked potentials (VEPs) to measure color discrimination thresholds and to plot color discrimination ellipses (MacAdam, 1942). Five normal trichromats (24.5 ± 2.6 years-old) were monocularly tested. Stimuli consisted of sinusoidal isoluminant chromatic gratings made from chromaticity pairs located along four different color directions radiating from one reference point of the CIE 1976 chromaticity diagram (u′ = 0.225; v′ = 0.415). Heterochromatic flicker photometry (HFP) was used to obtain the isoluminance condition for every subject and for all chromaticity pairs. VEPs were elicited using two cycles per degree grating stimuli at three different temporal configurations: transient, onset (300 ms)/offset (700 ms), 1 Hz fundamental frequency; steady-state, onset (50 ms)/offset (50 ms), 10 Hz fundamental frequency; and steady-state pattern reversal at 5 Hz fundamental frequency (10 Hz phase reversal). VEP amplitude was measured using transient VEP N1-P1 components and steady state VEP first (10 Hz) and second (20 Hz) harmonics. VEP amplitude was plotted as a function of chromatic distance in the CIE 1976 color space and the data points were extrapolated to zero amplitude level to obtain chromatic discrimination thresholds. The results were compared with psychophysical measurements performed using the same stimulus configurations and with the pseudoisochromatic method of Mollon-Reffin (one-way ANOVA). For all subjects and all stimulation methods, the ellipses showed small sizes, low ellipticities, and were vertically oriented. Despite some consistent differences in the results obtained with different procedures, there was no statistical difference between ellipses obtained electrophysiologically and psychophysically. For steady state VEPs, ellipses obtained from second harmonic amplitudes were larger and more elongated in the tritan direction than those obtained with first harmonic amplitudes.

Impulse response functions (IRFs) were obtained from two-pulse detection thresholds using isoluminant stimuli that produced increments or decrements in S-cone excitation. The pulses were chromatically modulated at constant luminance (based on 18 Hz heterochromatic flicker photometry). Chromatic stimuli were presented as a Gaussian patch (±1 SD = 2.3°) in one of four quadrants around a central fixation cross on a CRT screen. Each of the two pulses (6.67 ms) was separated by an inter-stimulus interval (ISI) from 20 to 360 ms. Chromaticity of the pulses was changed from the equal-energy white of the background to a bluish or yellowish color along individually determined tritan lines (based on color matching under strong S-cone adaptation from a 420 nm background superimposed in Maxwellian view). Chromatic detection thresholds were determined by a four-alternative forced-choice method with staircases for each ISI interleaved in each session. Measurements were repeated in at least four sessions for each observer. IRFs were calculated by varying four parameters of an exponentially-damped sinewave. Both S-cone increment and decrement IRFs are characterized by a single excitatory phase and a much longer time course compared with IRFs derived for luminance modulation using the same apparatus and observers. S-cone increment IRFs are faster than S-cone decrement IRFs; the time to peak amplitude of S-cone increment and decrement IRFs is 50–70 and 100–120 ms, respectively. These results were used to derive the temporal contrast sensitivity for human observers of putative ON- and OFF-channels carrying signals from S-cones.

In the natural environment, color discriminations are made within a rich context of spatial and temporal variation. In classical laboratory methods for studying chromatic discrimination, there is typically a border between the test and adapting fields that introduces a spatial chromatic contrast signal. Typically, the roles of spatial and temporal contrast on chromatic discrimination are not assessed in the laboratory approach. In this study, S-cone discrimination was measured using stimulus paradigms that controlled the level of spatio-temporal S-cone contrast between the tests and adapting fields. The results indicate that S-cone discrimination of chromaticity differences between a pedestal and adapting surround is equivalent for stimuli containing spatial, temporal or spatial-and-temporal chromatic contrast between the test field and the surround. For a stimulus condition that did not contain spatial or temporal contrast, the visual system adapted to the pedestal instead of the surround. The data are interpreted in terms of a model consistent with primate koniocellular pathway physiology. The paradigms provide an approach for studying the effects of spatial and temporal contrast on discrimination in natural scenes.

Under dichoptic viewing conditions, rivalrous gratings that differ in both color and form can give the percept of the color from one eye in part of the form in the other eye. This study examined the afterimage following such misbinding of color to form. The first experiment established that afterimages of the misbound percept were seen. Two possible mechanisms for the misbound afterimage are (1) persisting retinal representations that are rivalrous and subsequently resolved to give misbinding, as during rivalrous viewing, and (2) a persisting response from a central neural representation of the misbound percept with the form from one eye and color from the other eye. The results support afterimage formation from a central representation of the misbound percept, not from resolution of rivalrous monocular representations.

An open question in color rivalry is whether alternation between two colors is caused by a difference in receptoral stimulation or a difference in the neural representation of color appearance. This question was examined with binocular rivalry between physically identical lights that differed in appearance due to chromatic induction. Perceptual alternation was measured between gratings of the same chromaticity; each one was presented within a different patterned surround that caused the gratings, one to each eye, to appear unequal in hue because of chromatic induction. The gratings were presented dichoptically with binocular disparity so the rivalrous gratings appeared in front of the surround. Perceptual alternation in hue was found for the two physically identical chromaticities. Stereoscopic depth also was perceived, corroborating binocular neural combination despite color rivalry (Treisman, 1962). The results show that color rivalry is resolved after color-appearance shifts caused by chromatic context, and that color rivalry does not require competing unequal cone excitations from the rivalrous stimuli.

Luminance signals mediated by the magnocellular (MC) pathway play an important role in vernier tasks. MC ganglion cells show a phase advance in their responses to sinusoidal stimuli with increasing contrast due to contrast gain control mechanisms. If the phase information in MC ganglion cell responses were utilized by central mechanisms in vernier tasks, one might expect systematic errors caused by the phase advance. This systematic error may contribute to the contrast paradox phenomenon, where vernier performance deteriorates, rather than improves, when only one of the target pair increases in contrast. Vernier psychometric functions for a pair of gratings of mismatched contrast were measured to seek such misestimation. In associated electrophysiological experiments, MC and parvocellular (PC) ganglion cells' responses to similar stimuli were measured to provide a physiological reference. The psychophysical experiments show that a high-contrast grating is perceived as phase advanced in the drift direction compared to a low-contrast grating, especially at a high drift rate (8 Hz). The size of the phase advance was comparable to that seen in MC cells under similar stimulus conditions. These results are consistent with the MC pathway supporting vernier performance with achromatic gratings. The shifts in vernier psychometric functions were negligible for pairs of chromatic gratings under the conditions tested here, consistent with the lack of phase advance both in responses of PC ganglion cells and in frequency-doubled chromatic responses of MC ganglion cells.

Two experiments explore the color perception of objects in complex scenes. The first experiment examines the color perception of objects across variation in surface gloss. Observers adjusted the color appearance of a matte sphere to match that of a test sphere. Across conditions we varied the body color and glossiness of the test sphere. The data indicate that observers do not simply match the average light reflected from the test. Indeed, the visual system compensates for the physical effect of varying the gloss, so that appearance is stabilized relative to what is predicted by the spatial average. The second experiment examines how people perceive color across locations on an object. We replaced the test sphere with a soccer ball that had one of its hexagonal faces colored. Observers were asked to adjust the match sphere have the same color appearance as this test patch. The test patch could be located at either an upper or lower location on the soccer ball. In addition, we varied the surface gloss of the entire soccer ball (including the test patch). The data show that there is an effect of test patch location on observers' color matching, but this effect is small compared to the physical change in the average light reflected from the test patch across the two locations. In addition, the effect of glossy highlights on the color appearance of the test patch was consistent with the results from Experiment 1.