Cézar A. Saito

Morphology and physiology of primate M- and P-cells

Catarrhines and platyrrhines, the so-called Old- and New-World anthropoids, have different cone photopigments. Postreceptoral mechanisms must have co-evolved with the receptors to provide trichromatic color vision, and so it is important to compare postreceptoral processes in these two primate groups, both from anatomical and physiological perspectives. The morphology of ganglion cells has been studied in the retina of catarrhines such as the diurnal and trichromatic Macaca, as well as platyrrhines such as the diurnal, di- or trichromatic Cebus, and the nocturnal, monochromatic Aotus. Diurnal platyrrhines, both di- and trichromats, have ganglion cell classes very similar to those found in catarrhines: M (parasol), P (midget), small-field bistratified, and several classes of wide-field ganglion cells. In the fovea of all diurnal anthropoids, P-cell dendritic trees contact single midget bipolars, which contact single cones. The Aotus retina has far fewer cones than diurnal species, but M- and P-cells are similar to those in diurnal primates although of larger size. As in diurnal anthropoids, in the Aotus, the majority of midget bipolar cells, found in the central 2 mm of eccentricity, receive input from a single cone and the sizes of their axon terminals match the sizes of P-cell dendritic fields in the same region. The visual responses of retinal ganglion cells of these species have been studied using single-unit electrophysiological recordings. Recordings from retinal ganglion cells in Cebus and Aotus showed that they have very similar properties as those in the macaque, except that P-cells of mono- and dichromatic animals lack cone opponency. Whatever the original role of the M- and P-cells was, they are likely to have evolved prior to the divergence of catarrhines and platyrrhines. M- and P-cell systems thus appear to be strongly conserved in the various primate species. The reasons for this may lie in the roles of these systems for both achromatic and chromatic vision.

Visual impairment on dentists related to occupational mercury exposure

A detailed assessment of visual function was obtained in subjects with low-level occupational mercury exposure by measuring hue saturation thresholds and contrast sensitivity functions for luminance and chromatic modulation. General practice dentists (n=15) were compared to age-matched healthy controls (n=13). Color discrimination estimated by the area of Mac Adam ellipses was impaired, showing diffuse discrimination loss. There was also reduction of contrast sensitivity for luminance and chromatic (red-green and blue-yellow) modulation, in all tested spatial frequencies. Low concentrations of urinary mercury (1.97±1.61μg/g creatinine) were found in the dentists group. Color discrimination as well as contrast sensitivity function, assessed psychophysically, constitutes a sensitive indicator of subtle neurotoxic effect of elemental mercury exposure.

Cone selective adaptation influences L- and M-cone driven signals in electroretinography and psychophysics.

To assess the influence of selective adaptation of long (L) and middle (M) wavelength sensitive cones with electroretinography (ERG) and psychophysics, a novel adaptation procedure was developed, which comprises a selective and quantifiable change in the state of adaptation in the different cone types. One adaptation condition was used as a reference. In four additional conditions, the M-cones or the L-cones were selectively adapted, so that they absorbed either more or less photons. At each of these five states of adaptation, the ERG response amplitudes to 30Hz L- and to M-cone selective stimuli were measured. Furthermore, the psychophysical sensitivities to L- and M-cone selective stimuli were measured at different temporal frequencies. In subjects with normal color vision, adaptation can have a strong influence on the L- and M-cone driven response amplitudes in the ERG and on both the L- and the M-cone sensitivities in the psychophysical luminance channel. As a result, the L- to M-cone ERG and psychophysical ratios can change dramatically at the different states of adaptation. The cone sensitivity thresholds and the L- to M-cone sensitivity ratio in the psychophysical chromatic channel are about unity at all states of adaptation, suggesting the presence of a compensatory mechanism. In dichromats, the responses and sensitivities to stimulation of the absent cone type were generally small at all states of adaptation. But, with reddish backgrounds residual ERG responses and residual psychophysical sensitivities were observed, indicating the presence of either a robust rod driven signal or an additional adaptation mechanisms that are not cone driven and that have not been described before.

Twelve chromatically opponent ganglion cell types in turtle 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.

Amplitude of the transient visual evoked potential (tVEP) as a function of achromatic and chromatic contrast : Contribution of different visual pathways

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 degrees 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.

The topography of cone photoreceptors in the retina of a diurnal rodent, the agouti (Dasyprocta aguti)

The presence, density distribution, and mosaic regularity of cone types were studied in the retina of the diurnal agouti, Dasyprocta aguti. Longwave-sensitive (L-) and shortwave-sensitive (S-) cones were detected by antibodies against the respective cone opsins. L- and S-cones were found to represent around 90 and 10% of the cone population, respectively. There was no evidence for L- and S-opsin coexpression in agouti cones. L-cone densities were highest, up to 14,000/mm2, along a horizontal visual streak located about 2-3 mm dorsal to the optic nerve, and the L-cone distribution showed a dorsoventral asymmetry with higher densities in ventral (about 10,000/mm2) than in dorsal (about 4000/mm2) retinal regions. This L-cone topography parallels the agoutis ganglion cell topography. S-cones had a peak density of 1500-2000/mm2 in the central retinal region but did not form a visual streak. Their distribution also showed a dorsoventral asymmetry with densities around 600/mm2 in dorsal and around 1000/mm2 in ventral retinal regions. The patterning of cone arrays was assessed by the density recovery profile analysis. At all eccentricities evaluated, the S-cone mosaic less efficiently packed than the L-cone mosaic. Rod densities ranged from 47,000/mm2 in peripheral to 64,000/mm2 in central retina, and rod:cone ratios were 4:1-9:1. The comparatively low rod density and high cone proportion appear well adapted to the diurnal lifestyle of the agouti.

Morphology of horizontal cells in the retina of the capuchin monkey, Cebus apella: How many horizontal cell classes are found in dichromatic primates?

The morphology of horizontal cells was studied in the retina of dichromatic capuchin monkeys, Cebus apella. The cells were labeled with the carbocyanine dye, 1,1′,dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI), and the labeling was then photoconverted to a stable product by using a diaminobenzidine reaction. The sizes of cell body, dendritic field, and axon terminal, as well as the number of dendritic clusters and cone convergence, were measured at increasing distance from the fovea. Three distinct morphological classes of horizontal cells were identified. Their dendritic and axonal morphology resembles those of H1, H2, and H3 cells described in trichromatic primates. The size of the cell bodies, dendritic fields, and axon terminals of all cell classes increases towards retinal periphery. H3 cells have larger dendritic fields and more dendritic clusters than H1 cells. All labeled horizontal cells located in selected patches of retina were further analyzed to quantify the differences between H1 and H3 cells. H1 cells have smaller dendritic field area, smaller total length of primary dendrites, more dendritic branching points, and larger fractal dimension than H3 cells. We have distinguished H1 and H3 cells based solely in morphological criteria. Their physiology should be further analyzed with detail, but their presence in both dichromatic and trichromatic primates suggests that neither of them have a specialized role in the red‐green color opponent channel of color vision. J. Comp. Neurol. 443:105–123, 2002.

Normal and dichromatic color discrimination measured with transient visual evoked potential.

It would be informative to have an electrophysiological method to study, in an objective way, the effects of mercury exposure and other neurotoxics on human color vision performance. The purpose of the present work was to study human color discrimination by measuring chromatic difference thresholds with visual evoked potential (VEP). Six young normal trichromats (24 +/- 1 years old) and one deutan (26 years old) were tested. The stimuli consisted of sinusoidal isoluminant chromatic gratings made from chromaticity pairs located along four different color directions centered on two reference points. Heterochromatic flicker photometry (HFP) protocol was used to obtain the isoluminance condition for every subject and for all chromaticity pairs. Spatial frequency was 2 cycles/deg. Presentation mode comprised onset (300 ms)/offset (700 ms) periods. As previously described, we found a negative deflection in the VEP which was related to the chromatic difference: as chromatic difference increased, amplitude increased and latency decreased. VEP response amplitude was plotted against distance in the CIE 1976 color space between the grating chromaticities and fitted with a regression line. We found color thresholds by extrapolating the fitting to null amplitude values. The thresholds were plotted in the CIE 1976 color space as MacAdam ellipses. In normal trichromats the ellipses had small size, low ellipticity, and were vertically oriented. In the deutan subject, the ellipses had large size, high ellipticity, and were oriented towards the deutan copunctal locus. The VEP thresholds were similar to those obtained using grating stimuli and psychophysical procedures, however smaller than those obtained using pseudoisochromatic stimuli (Mollon-Reffin method). We concluded that transient VEP amplitude as a function of contrast can be reliably used in objective studies of chromatic discrimination performance in normal and altered human subjects.

Horizontal cells in the retina of a diurnal rodent, the agouti ( Dasyprocta aguti )

The morphology and distribution of normally placed and displaced A horizontal cells were studied in the retina of a diurnal hystricomorph rodent, the agouti Dasyprocta aguti. Cells were labeled with anti-calbindin immunocytochemistry. Dendritic-field size reaches a minimum in the visual streak, of about 9,000 microm(2), and increases toward the retinal periphery both in the dorsal and ventral regions. There is a dorsoventral asymmetry, with dorsal cells being larger than ventral cells at equal distances from the streak. The peak value for cell density of 281 +/- 28 cells/mm(2) occurs in the center of the visual streak, decreasing toward the dorsal and ventral retinal periphery, paralleling the increase in dendritic-field size. Along the visual streak, the decline in cell density is less pronounced, remaining between 100-200 cells/mm(2) in the temporal and nasal periphery. Displaced horizontal cells are rare and occur in the retinal periphery. They tend to be smaller than normally placed horizontal cells in the ventral region, whilst no systematic difference was observed between the two cell groups in the dorsal region. Mosaic regularity was studied using nearest-neighbor analysis and the Ripley function. When mosaic regularity was determined removing the displaced horizontal cells, there was a slight increase in the conformity ratio, but the bivariate Ripley function indicated some repulsive dependence between the two mosaics. Both results were near the level of significance. A similar analysis performed in the capybara retina, a closely related hystricomorph rodent bearing a higher density of displaced horizontal cells than found in the agouti, suggested spatial independence between the two mosaics, normally placed versus displaced horizontal cells.

Voronoi analysis uncovers relationship between mosaics of normally placed and displaced amacrine cells in the thraira retina

Although neuronal dynamics is to a high extent a function of synapse strength, the spatial distribution of neurons is also known to play an important role, which is evidenced by the topographical organization of the main stations of the visual system: retina, lateral geniculate nucleus, and cortex. The coexisting systems of normally placed and displaced amacrine cells in the vertebrate retina provide interesting examples of retinotopic spatial organization. However, it is not clear whether these two systems are spatially interrelated or not. The current work applies two mathematical-computational methods-a new method involving Voronoi diagrams for local density quantification and a more traditional approach, the Ripley K function-in order to characterize the mosaics of normally placed and displaced amacrine cells in the retina of Hoplias malabaricus and search for possible spatial relationships between these two types of mosaics. The results obtained by the Voronoi local density analysis suggest that the two systems of amacrine cells are spatially interrelated through nearly constant local density ratios.