Sex-related differences in chromatic sensitivity.

Generally women are believed to be more discriminating than men in the use of color names and this is often taken to imply superior color vision. However, if both X-chromosome linked color deficient males (8%) and females (<1%) as well as heterozygote female carriers (15%) are excluded from comparisons, then differences between men and women in red-green (RG) color discrimination have been reported as not being significant (e.g., Pickford, 1944; Hood et al., 2006). We re-examined this question by assessing the performance of 150 males and 150 females on the color assessment and diagnosis (CAD) test (Rodriguez-Carmona et al., 2005). This is a sensitive test that yields small color detection thresholds. The test employs direction-specific, moving, chromatic stimuli embedded in a background of random, dynamic, luminance contrast noise. A four-alternative, forced-choice procedure is employed to measure the subject's thresholds for detection of color signals in 16 directions in color space, while ensuring that the subject cannot make use of any residual luminance contrast signals. In addition, we measured the Rayleigh anomaloscope matches in a subgroup of 111 males and 114 females. All the age-matched males (30.8 +/- 9.7) and females (26.7 +/- 8.8) had normal color vision as diagnosed by a battery of conventional color vision tests. Females with known color deficient relatives were excluded from the study. Comparisons between the male and female groups revealed no significant differences in anomaloscope midpoints (p = 0.709), but a significant difference in matching ranges (p = 0.040); females on average tended to have a larger mean range (4.11) than males (3.75). Females also had significantly higher CAD thresholds than males along the RG (p = 0.0004), but not along the yellow-blue (YB) discrimination axis. The differences between males and females in RG discrimination may be related to the heterozygosity in X-linked cone photo pigment expression common among females.

M-cone opsin gene number does not correlate with variation in L/M-cone sensitivity.

Molecular genetic studies demonstrate that the human cone opsin gene array on the q-arm of the X-chromosome typically consists of one long-wave-sensitive (L) cone opsin gene and from one to several middle-wave-sensitive (M) cone opsin genes. Although the presence of the single L-cone opsin gene and at least one M-cone opsin gene is essential for normal red-green colour discrimination, the function of the additional M-cone opsin genes is still unclear. To investigate whether any variations in phenotype correlate with differences in the number of M-cone opsin genes, we selected 13 normal trichromat males, for whom four independent molecular techniques have exactly determined their number of M-cone opsin genes, ranging from one to four. Their phenotype was characterized by estimating their foveal L- to M-cone ratio from heterochromatic flicker photometric (HFP) thresholds, by measuring the wavelength corresponding to their 'unique yellow', and by determining their L- and M-cone modulation thresholds (CMTs). No correlation was found between these psychophysical measures and the number of M-cone opsin genes. Although, we found a reasonably good correlation between the L/M-cone ratios based on HFP and on CMT, we did not find any correlation between the estimated L/M-cone ratios and the settings of 'unique yellow'. Our results accord with previous molecular genetic studies that suggest that only the first two genes in the X-linked opsin gene array are expressed.

CNGA3 mutations in hereditary cone photoreceptor disorders.

We recently showed that mutations in the CNGA3 gene encoding the alpha-subunit of the cone photoreceptor cGMP-gated channel cause autosomal recessive complete achromatopsia linked to chromosome 2q11. We now report the results of a first comprehensive screening for CNGA3 mutations in a cohort of 258 additional independent families with hereditary cone photoreceptor disorders. CNGA3 mutations were detected not only in patients with the complete form of achromatopsia but also in incomplete achromats with residual cone photoreceptor function and (rarely) in patients with evidence for severe progressive cone dystrophy. In total, mutations were identified in 53 independent families comprising 38 new CNGA3 mutations, in addition to the 8 mutations reported elsewhere. Apparently, both mutant alleles were identified in 47 families, including 16 families with presumed homozygous mutations and 31 families with two heterozygous mutations. Single heterozygous mutations were identified in six additional families. The majority of all known CNGA3 mutations (39/46) are amino acid substitutions compared with only four stop-codon mutations, two 1-bp insertions and one 3-bp in-frame deletion. The missense mutations mostly affect amino acids conserved among the members of the cyclic nucleotide gated (CNG) channel family and cluster at the cytoplasmic face of transmembrane domains (TM) S1 and S2, in TM S4, and in the cGMP-binding domain. Several mutations were identified recurrently (e.g., R277C, R283W, R436W, and F547L). These four mutations account for 41.8% of all detected mutant CNGA3 alleles. Haplotype analysis suggests that the R436W and F547L mutant alleles have multiple origins, whereas we found evidence that the R283W alleles, which are particularly frequent among patients from Scandinavia and northern Italy, have a common origin.

Abnormal retinotopic representations in human visual cortex revealed by fMRI.

The representation of the visual field in early visual areas is retinotopic. The point-to-point relationship on the retina is therefore maintained on the convoluted cortical surface. Functional magnetic resonance imaging (fMRI) has been able to demonstrate the retinotopic representation of the visual field in occipital cortex of normal subjects. Furthermore, visual areas that are retinotopic can be identified on computationally flattened cortical maps on the basis of positions of the vertical and horizontal meridians. Here, we investigate abnormal retinotopic representations in human visual cortex with fMRI. We present three case studies in which patients with visual disorders are investigated. We have tested a subject who only possesses operating rod photoreceptors. We find in this case that the cortex undergoes a remapping whereby regions that would normally represent central field locations now map more peripheral positions in the visual field: In a human albino we also find abnormal visual cortical activity. Monocular stimulation of each hemifield resulted in activations in the hemisphere contralateral to the stimulated eye. This is consistent with abnormal decussation at the optic chiasm in albinism. Finally, we report a case where a lesion to white matter has resulted in a lack of measurable activity in occipital cortex. The activity was absent for a small region of the visual field, which was found to correspond to the subject's field defect. The cases selected have been chosen to demonstrate the power of fMRI in identifying abnormalities in the cortical representations of the visual field in patients with visual dysfunction. Furthermore, the experiments are able to show how the cortex is capable of modifying the visual field representation in response to abnormal input.

Motion perception at scotopic light levels.

Although the spatial and temporal properties of rod-mediated vision have been extensively characterized, little is known about scotopic motion perception. To provide such information, we determined thresholds for the detection and identification of the direction of motion of sinusoidal grating patches moving at speeds from 1 to 32 deg/s, under scotopic light levels, in four different types of observers: three normals, a rod monochromat (who lacks all cone vision), an S-cone monochromat (who lacks M- and L-cone vision), and four deuteranopes (who lack M-cone vision). The deuteranopes, whose motion perception does not differ from that of normals, allowed us to measure rod and L-cone thresholds under silent substitution conditions and to compare directly the perceived velocity for moving stimuli detected by either rod or cone vision at the same light level. We find, for rod as for cone vision, that the direction of motion can be reliably identified very near to detection threshold. In contrast, the perceived velocity of rod-mediated stimuli is reduced by approximately 20% relative to cone-mediated stimuli at temporal frequencies below 4 Hz and at all intensity levels investigated (0.92 to -1.12 log cd m(-2)). Most likely, the difference in velocity perception is distal in origin because rod and cone signals converge in the retina and further processing of their combined signals in the visual cortex is presumably identical. To account for the difference, we propose a model of velocity, in which the greater temporal averaging of rod signals in the retina leads to an attenuation of the motion signal in the detectors tuned to high velocities.

Mutations in the CNGB3 gene encoding the beta-subunit of the cone photoreceptor cGMP-gated channel are responsible for achromatopsia (ACHM3) linked to chromosome 8q21.

Achromatopsia is an autosomal recessive disorder featuring total colour blindness, photophobia, reduced visual acuity and nystagmus. While mutations in the CNGA3 gene on chromosome 2q11 are responsible for achromatopsia in a subset of patients, previous linkage studies have localized another achromatopsia locus, ACHM3, on chromosome 8q21. Using achromatopsia families in which CNGA3 mutations have been excluded, we refined the ACHM3 locus to a 3.7 cM region enclosed by markers D8S1838 and D8S273. Two yeast artificial chromosome (YAC) contigs covering nearly the entire ACHM3 interval were constructed. Database searches with YAC content sequences identified two overlapping high throughput genomic sequencing phase (HTGS) entries which contained sequences homologous to the murine cng6 gene encoding the putative beta-subunit of the cone photoreceptor cGMP-gated channel. Using RT-PCR and RACE, we identified and cloned the human cDNA homologue, designated CNGB3, which encodes an 809 amino acid polypeptide. Northern blot analysis revealed a major transcript of approximately 4.4 kb specifically expressed in the retina. The human CNGB3 gene consists of 18 exons distributed over approximately 200 kb of genomic sequence. Analysis of the CNGB3 gene in achromats revealed six different mutations including a missense mutation (S435F), two stop codon mutations (R203X and E336X), a 1 bp and an 8 bp deletion (1148delC and 819-826del) and a putative splice site mutation of intron 13. The 1148delC mutation was identified recurrently in several families, and in total was present on 11 of 22 disease chromosomes segregating in our families.

The spectral sensitivities of the middle- and long-wavelength-sensitive cones derived from measurements in observers of known genotype.

The spectral sensitivities of middle- (M-) and long- (L-) wavelength-sensitive cones have been measured in dichromats of known genotype: M-cone sensitivities in nine protanopes, and L-cone sensitivities in 20 deuteranopes. We have used these dichromat cone spectral sensitivities, along with new luminous efficiency determinations, and existing spectral sensitivity and color matching data from normal trichromats, to derive estimates of the human M- and L-cone spectral sensitivities for 2 and 10 degrees dia. central targets, and an estimate of the photopic luminosity function [V(lambda)] for 2 degrees dia. targets, which we refer to as V(2)*(lambda). These new estimates are consistent with dichromatic and trichromatic spectral sensitivities and color matches.

Tritanopic color matches and the middle- and long-wavelength-sensitive cone spectral sensitivities.

Tritanopic color matches (i.e. matches that depend on the middle- (M) and long- (L), but not short- (S) wavelength-sensitive cones) were made between two half-fields: one illuminated by either a 405 or a 436 nm Hg spectral line; the other by a light of variable wavelength and radiance. Our purpose was to test between rival M- and L-cone spectral sensitivities, which should predict the tritanopic matches. The observers were tritanopes, in whom functioning S-cones are lacking, or normal trichromats, in whom artificial tritanopia was induced by a strong, violet adapting field. The wavelengths found to match the 405 and 436 nm lights agreed poorly with those predicted by the cone spectral sensitivities of Smith and Pokorny (1975) [Vision Research, 15, 161], while the 405 nm matching wavelength agreed poorly with that predicted by Stockman, MacLeod and Johnson (1993) [Journal of the Optical Society of America, A10, 2491]. Both matching wavelengths agreed well, however, with the predictions of the Stockman and Sharpe (2000) [Vision Research] M- and L-cone spectral sensitivities, which lie within the range of measured matches.

Asymmetries in the time-course of chromatic adaptation and the significance of contrast.

The time-course of chromatic adaptation was determined as a function of the spectral content of the adaptation-light and of image-contrast. The adaptation-lights varied along the chromatic cardinal axes or one of their intermediate axes in an equiluminant plane in colour-space. We found an asymmetry in the initial time-course of adaptation (0.2-10 s) in that adaptation to middle-wavelength light was significantly faster than adaptation to short- and long-wavelength light. The asymmetry was only observed in the presence of a spatially complex pattern. It was fully supported by luminance and chromatic contrast and present under haploscopic stimulus conditions.

L/M cone ratios in human trichromats assessed by psychophysics, electroretinography, and retinal densitometry.

Estimates of the relative numbers of long-wavelength-sensitive (L) and middle-wavelength-sensitive (M) cones vary considerably among normal trichromats and depend significantly on the nature of the experimental method employed. Here we estimate L/M cone ratios in a population of normal observers, using three psychophysical tasks-detection thresholds for cone-isolating stimuli at different temporal frequencies, heterochromatic flicker photometry, and cone contrast ratios at minimal flicker perception--as well as flicker electroretinography and retinal densitometry. The psychophysical tasks involving high temporal frequencies, specifically designed to tap into the luminance channel, provide average L/M cone ratios that significantly differ from unity with large interindividual variation. In contrast, the psychophysical tasks involving low temporal frequencies, chosen to tap into the red-green chromatic channel, provide L/M cone ratios that are always close to unity. L/M cone ratios determined from electroretinographic recordings or from retinal densitometry correlate with those determined from the high-temporal-frequency tasks. These findings suggest that the sensitivity of the luminance channel is directly related to the relative densities of the L and the M cones and that the red-green chromatic channel introduces a gain adjustment to compensate for differences in L and M cone signal strength.

Rod pathways: the importance of seeing nothing.

Anatomical and physiological studies of the mammalian retina have revealed two primary pathways available for the transmission of rod signals to the ganglion cells: one via ON rod bipolars, amacrine II cells, and ON and OFF cone bipolars, which is exquisitely designed for the transmission of single-photon absorption events; and a second via rod-cone gap junctions, and ON and OFF cone bipolars, which is designed for the transmission of multiple photon-absorption events at higher light levels. Psychophysical and electroretinographic (ERG) studies in normal observers and in two rare types of observer, who are devoid of either rod or cone function, support an analogous duality in the human visual system, the clearest signature of which is a loss of flicker visibility and ERG amplitude at frequencies near 15 Hz that results from destructive interference between sensitive 'slow' and insensitive 'fast' rod signals. The slow rod signal is most probably derived from the ON rod bipolar pathway and the fast signal from the rod-cone gap junction and cone pathways. Evidence has emerged recently for a third, insensitive rod pathway between rods and OFF cone bipolars, but it has so far only been observed clearly in rodents.

The spectral sensitivity of the human short-wavelength sensitive cones derived from thresholds and color matches.

We used two methods to estimate short-wave (S) cone spectral sensitivity. Firstly, we measured S-cone thresholds centrally and peripherally in five trichromats, and in three blue-cone monochromats, who lack functioning middle-wave (M) and long-wave (L) cones. Secondly, we analyzed standard color-matching data. Both methods yielded equivalent results, on the basis of which we propose new S-cone spectral sensitivity functions. At short and middle-wavelengths, our measurements are consistent with the color matching data of Stiles and Burch (1955, Optica Acta, 2, 168-181; 1959, Optica Acta, 6, 1-26), and other psychophysically measured functions, such as pi 3 (Stiles, 1953, Coloquio sobre problemas opticos de la vision, 1, 65-103). At longer wavelengths, S-cone sensitivity has previously been over-estimated.

Direct visual resolution of gene copy number in the human photopigment gene array.

PURPOSE: To visualize by direct fluorescent in situ hybridization the entire human visual pigment gene array on single X-chromosome fibers and to compare the results with values obtained by other molecular techniques. METHODS: The size of the opsin gene array on the X-chromosome in eight male subjects was investigated by (i) direct visual in situ hybridization (DIRVISH) on elongated DNA fibers: (ii) quantitation of genomic restriction fragments after Southern blot hybridization; (iii) quantitation of restriction fragment length polymorphism after PCR amplification (PCR/RFLP), and (iv) sizing of NotI fragments by pulsed field gel electrophoresis and Southern blot detection. Each male subject's color vision was assessed by Rayleigh matches on a Nagel Type 1 anomaloscope. RESULTS: The number of genes resolved by the DIRVISH protocol, which ranges from 1 to 6, agrees exactly with the gene array sizes obtained in the same male subjects from pulsed field gel electrophoresis, but differs from the estimates derived from the commonly used indirect Southern blot hybridization and PCR/RFLP quantitation methods. In particular, the PCR/RFLP method overestimates the copy number in all but the smallest arrays. CONCLUSIONS: Visualization of the X-chromosome opsin gene array by DIRVISH provides a new, direct method for obtaining exact copy numbers and helps to resolve the controversy about the range and the average visual pigment gene number in the human population in favor of smaller average array sizes.

Cone signal contributions to electroretinograms [correction of electrograms] in dichromats and trichromats.

PURPOSE: To find out how the different cone types contribute to the electroretinogram (ERG) by quantifying the contribution of the signal pathways originating in the long (L-) and the middle (M-) wavelength-sensitive cones to the total ERG response amplitude and phase. METHODS: ERG response amplitudes and phases were measured to cone-isolating stimuli and to different combinations of L- and M-cone modulation. Conditions were chosen to exclude any contribution of the short wavelength-sensitive (S-) cones. The sensitivity of the ERG to the L and the M cones was defined as the cone contrast gain. RESULTS: In the present paper, a model is provided that describes the ERG contrast gains and ERG thresholds in dichromats and color normal trichromats. For the X-chromosome-linked dichromats, the contrast gains of only one cone type (either the L or the M cones) sufficed to describe the ERG thresholds for all stimulus conditions. Data suggest that the M-cone contrast gains of protanopes are larger than the L-cone contrast gains of deuteranopes. The response thresholds of the trichromats are modeled by assuming a vector summation of signals originating in the L and the M cones. Their L- and M-cone contrast gains are close to a linear interpolation of the data obtained from the dichromats. Nearly all trichromats had larger L- than M-cone contrast gains. Data from a large population of trichromats were examined to study the individual variations in cone weightings and in the phases of the cone pathway responses. CONCLUSIONS: The data strongly suggest that the missing cone type in dichromats is replaced by the remaining cone type. The mean L-cone to M-cone weighting ratio in trichromats was found to be approximately 4:1. But there is a substantial interindividual variability between trichromats. The response phases of the L- and the M-cone pathways can be reliably quantified using the response phases to the cone-isolating stimuli or using a vector addition of L- and M-cone signals.

Selective color constancy deficits after circumscribed unilateral brain lesions.

The color of an object, when part of a complex scene, is determined not only by its spectral reflectance but also by the colors of all other objects in the scene (von Helmholtz, 1886; Ives, 1912; Land, 1959). By taking global color information into account, the visual system is able to maintain constancy of the color appearance of the object, despite large variations in the light incident on the retina arising from changes in the spectral content of the illuminating light (Hurlbert, 1998; Maloney, 1999). The neural basis of this color constancy is, however, poorly understood. Although there seems to be a prominent role for retinal, cone-specific adaptation mechanisms (von Kries, 1902; Pöppel, 1986; Foster and Nascimento, 1994), the contribution of cortical mechanisms to color constancy is still unclear (Land et al., 1983; D'Zmura and Lennie, 1986). We examined the color perception of 27 patients with defined unilateral lesions mainly located in the parieto-temporo-occipital and fronto-parieto-temporal cortex. With a battery of clinical and specially designed color vision tests we tried to detect and differentiate between possible deficits in central color processing. Our results show that color constancy can be selectively impaired after circumscribed unilateral lesions in parieto-temporal cortex of the left or right hemisphere. Five of 27 patients exhibited significant deficits in a color constancy task, but all of the 5 performed well in color discrimination or higher-level visual tasks, such as the association of colors with familiar objects. These results indicate that the computations underlying color constancy are mediated by specialized cortical circuitry, which is independent of the neural substrate for color discrimination and for assigning colors to objects.

Evaluation of the Night Vision Spectacles on patients with impaired night vision.

BACKGROUND: The Night Vision Spectacles (NiViS) were developed by a consortium of European companies to assist individuals who suffer from impaired night vision. They consist of a head-mounted video camera (input) and binocular displays (output) connected to a portable computer processor, which uses an algorithm to enhance the luminance and contrast of the video image. METHODS: Eighteen patients with impaired night vision were tested, including those with retinitis pigmentosa (7), Usher syndrome (2), fundus albipunctatus (1) and complete (4) and incomplete (4) congenital stationary night blindness. Normal trichromats (3) and typical, complete achromats (2) acted as controls. A battery of tests assessed: visual acuity at 5 m (projection unit) and 1 m (chart) and at high and low contrasts; contrast sensitivity; absolute and increment threshold; the influence of glare; contrast motion detection; and hand-eye performance. The tests were performed, with and without the NiViS, at three adaptation levels: low scotopic (10(-3) cd/m2), high scotopic (10(-2) cd/m2) and mesopic (10(-1) cd/m2). RESULTS: At the low and high scotopic levels, the majority of patients showed improved performance on the visual acuity, contrast sensitivity and motion contrast tests with the NiViS. At the mesopic level, the advantage with the NiViS was greatly reduced, but still present for contrast sensitivity. CONCLUSION: Patients with impaired night vision can benefit from the NiViS when performing tasks involving contrast and motion perception. Those with normal visual fields and retaining good photopic vision will benefit more than those with constricted visual fields and impaired cone vision. Recommendations regarding desirable improvements of the NiViS and suitability for the individual patient are given.

Color signals in human motion-selective cortex.

The neural basis for the effects of color and contrast on perceived speed was examined using functional magnetic resonance imaging (fMRI). Responses to S cone (blue-yellow) and L + M cone (luminance) patterns were measured in area V1 and in the motion area MT+. The MT+ responses were quantitatively similar to perceptual speed judgments of color patterns but not to color detection measures. We also measured cortical motion responses in individuals lacking L and M cone function (S cone monochromats). The S cone monochromats have clear motion-responsive regions in the conventional MT+ position, and their contrast-response functions there have twice the responsivity of S cone contrast-response functions in normal controls. But, their responsivity is far lower than the normals' responsivity to luminance contrast. Thus, the powerful magnocellular input to MT+ is either weak or silent during photopic vision in S cone monochromats.

L, M and L-M hybrid cone photopigments in man: deriving lambda max from flicker photometric spectral sensitivities.

Using heterochromatic flicker photometry, we have measured the corneal spectral sensitivities of the X-chromosome-linked photopigments in 40 dichromats, 37 of whom have a single opsin gene in their tandem array. The photopigments encoded by their genes include: the alanine variant of the normal middle-wavelength sensitive photopigment, M(A180); the alanine and serine variants of the normal long-wavelength sensitive photopigment, L(A180) and L(S180); four different L-M hybrid or anomalous photopigments, L2M3(A180), L3M4(S180), L4M5(A180) and L4M5(S180); and two variants of the L-cone photopigment, encoded by genes with embedded M-cone exon two sequences, L(M2; A180) and L(M2; S180). The peak absorbances (lambda max) of the underlying photopigment spectra associated with each genotype were estimated by correcting the corneal spectral sensitivities back to the retinal level, after removing the effects of the macular and lens pigments and fitting a template of fixed shape to the dilute photopigment spectrum. Details of the genotype-phenotype correlations are summarized elsewhere (Sharpe, L. T., Stockman, A., Jägle, H., Knau, H., Klausen, G., Reitner, A. et al. (1998). J. Neuroscience, 18, 10053-10069). Here, we present the individual corneal spectral sensitivities for the first time as well as details and a comparison of three analyses used to estimate the lambda max values, including one in which the lens and macular pigment densities of each observer were individually measured.