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L T Sharpe

Publications and source records attributed to L T Sharpe.

At least 37 records · Page 2Linked to original sources
PubMed1999

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.

Blotting, Southern↗
PubMed1998

Red, green, and red-green hybrid pigments in the human retina: correlations between deduced protein sequences and psychophysically measured spectral sensitivities.

To analyze the human red, green, and red-green hybrid cone pigments in vivo, we studied 41 male dichromats, each of whose X chromosome carries only a single visual pigment gene (single-gene dichromats). This simplified arrangement avoids the difficulties of complex opsin gene arrays and overlapping cone spectral sensitivities present in trichromats and of multiple genes encoding identical or nearly identical cone pigments in many dichromats. It thus allows for a straightforward correlation between each observer's spectral sensitivity measured at the cornea and the amino acid sequence of his visual pigment. For each of the 41 single-gene dichromats we determined the amino acid sequences of the X-linked cone pigment as deduced from its gene sequence. To correlate these sequences with spectral sensitivities in vivo, we determined the Rayleigh matches to different red/green ratios for 29 single-gene dichromats and measured psychophysically the spectral sensitivity of the remaining green (middle wavelength) or red (long wavelength) cones in 37 single-gene dichromats. Cone spectral sensitivity maxima obtained from subjects with identical visual pigment amino acid sequences show up to a approximately 3 nm variation from subject to subject, presumably because of a combination of inexact (or no) corrections for variation in preretinal absorption, variation in photopigment optical density, optical effects within the photoreceptor, and measurement error. This variation implies that spectral sensitivities must be averaged over multiple subjects with the same genotype to obtain representative values for a given pigment. The principal results of this study are that (1) approximately 54% of the single-gene protanopes (and approximately 19% of all protanopes) possess any one of several 5'red-3'green hybrid genes that encode anomalous pigments and that would be predicted to produce protanomaly if present in anomalous trichromats; (2) the alanine/serine polymorphism at position 180 in the red pigment gene produces a spectral shift of approximately 2.7 nm; (3) for each exon the set of amino acids normally associated with the red pigment produces spectral shifts to longer wavelengths, and the set of amino acids normally associated with the green pigment produces spectral shifts to shorter wavelengths; and (4) changes in exons 2, 3, 4, and 5 from green to red are associated with average spectral shifts to long wavelengths of approximately 1 nm (range, -0.5 to 2.5 nm), approximately 3.3 nm (range, -0.5 to 7 nm), approximately 2.8 nm (range, -0.5 to 6 nm), and approximately 24.9 nm (range, 22.2-27.6 nm).

Adolescent↗
PubMed1998

Human rod monochromacy: linkage analysis and mapping of a cone photoreceptor expressed candidate gene on chromosome 2q11.

We have performed linkage analysis in eight families with rod monochromacy, an autosomal recessively inherited condition with complete color blindness. Significant linkage was found with markers located at the pericentromeric region of chromosome 2. A maximum lod score of 5.36 was obtained for marker D2S2333 at theta = 0.00. Mapping of meiotic breakpoints localized the disease gene between markers D2S2187 and D2S2229. Homozygosity for a number of subsequent markers indicating identity by descent was found in two families and provides evidence for a further refinement of the locus proximal to D2S373. This defines an interval of approximately 3 cM covering the ACHM2 locus for rod monochromacy. Radiation hybrid mapping of the CNGA3 gene encoding the alpha-subunit of the cGMP gated cation channel in human cone photoreceptors resulted in a maximum lod score of 16.1 with marker D2S2311 combined with a calculated physical distance of 6.19cR10,000. Screening of the CEPH YAC library and subsequent STS mapping indicated the physical order cen-D2S2222-D2S2175-(D2S2187/D2S2311)-qtel ofmarkers on 2q11 and showed that the CNGA3 gene maps most closely to D2S2187 and D2S2311. These data indicate that the CNGA3 gene maps within the critical interval of the ACHM2 locus for rod monochromacy and thus is a candidate gene for this disease.

Chromosome Breakage↗
PubMed1998

A computer-controlled system for measuring dark adaptation and other psychophysical functions.

BACKGROUND: Psychophysical thresholds, including dark-adaptation functions and increment threshold sensitivities, are useful for the early detection and diagnosis of visual pathologies. However, few instruments have been designed or adapted for their routine clinical measurement. METHODS: Here we describe an instrument prototype designed to meet this need, which we refer to as the PULS (Programmier-barer Universeller Licht-Stimulator). The instrument is computer-controlled and fully automated. It allows direct control over target location, luminance, size, duration and temporal profile and over background luminance and spectral composition. It also incorporates an efficient and statistically rigorous strategy for determining threshold. RESULTS: We present examples of psychophysical functions-dark-adaptation curves, increment threshold sensitivities, estimates of temporal summation in the dark and during the time course of dark adaptation-which have been measured by the PULS prototype in normal observers and clinical patients. CONCLUSIONS: The PULS instrument provides an automatic and efficient means of measuring dark adaptation and other psychophysical functions. It determines threshold by a more rigorous and faster method than is conventionally employed in clinical adaptometry.

Adolescent↗
PubMed1998

The contribution of color to visual memory in X-chromosome-linked dichromats.

We used a recognition memory paradigm to assess the visual memory of X-chromosome-linked dichromats for color images of natural scenes. The performance of 17 protanopes and 14 deuteranopes, who lack the second (red-green opponent) subsystem of color vision, but retain the primordial (yellow-blue opponent) subsystem, was compared with that of 36 color normal observers. During the presentation phase, 48 images of natural scenes were displayed on a CRT for durations between 50 and 1000 msec. Each image was followed by a random noise mask. Half of the images were presented in color and half in black and white. In the subsequent query phase, the same 48 images were intermixed with 48 new images and the subjects had to indicate which of the images they had already seen during the presentation phase. We find that the performance of the color normal observers increases with exposure duration. However, they perform 5-10% better for colored than for black and white images, even at exposure durations as short as 50 msec. Surprisingly, performance is not impaired for the dichromats, whose recognition performance is also better for colored than for black and white images. We conclude either that X-chromosome-linked dichromats may be able to compensate for their reduced chromatic information range when viewing complex natural scenes or that the chromatic information in most natural scenes, for the durations tested, is sufficiently represented by the surviving primordial color subsystem.

Adult↗
PubMed1998

Macular pigment densities derived from central and peripheral spectral sensitivity differences.

Estimates of the density spectrum of the macular pigment (Wyszecki G, Stiles WS. Color Science: Concepts and Methods. Quantitative Data and Formulas. 1st ed. New York: Wiley, 1967); (Vos JJ. Literature review of human macular absorption in the visible and its consequences for the cone receptor primaries. Institute for Perception. Soesterberg, The Netherlands, 1972) are partially based on the difference between central and peripheral spectral sensitivities, measured under conditions chosen to isolate a single cone class (Stiles WS. Madrid: Union Internationale de Physique Pure et Appliquée, 1953;1:65-103). Such derivations assume that the isolated spectral sensitivity is the same at both retinal locations, save for the intervening macular pigment. If this is true, then the type of cone class mediating detection should not influence the calculated difference spectrum. To test this assumption, we measured central and peripheral spectral sensitivities in a deuteranope, a protanope and a normal trichromat observer: (a) for short-wave sensitive (S-) cone detection; and (b) for long-wave sensitive (L-) cone detection (deuteranope), for middle-wave sensitive (M-) cone detection (protanope) or for both L- and M-cone detection (normal trichromat). The difference spectra determined for L- or M-cone detection deviate significantly from those measured for S-cone detection, at wavelengths below 450 nm. A theoretical analysis suggests that the discrepancies are owing, in part, to regional variation in the optical density of the cone pigments; and that such receptor variation cannot be ignored when deriving the standard density spectrum of the macular pigment.

Color Vision Defects↗
PubMed1998

Flicker cone electroretinogram in dichromats and trichromats.

To measure cone signal strengths in the flicker electroretinogram (ERG) of dichromats and trichromats, we developed a set of flickering stimuli (30 Hz), which excite the middle-wavelength-sensitive (M-) and long-wavelength-sensitive (L-) cones independently. ERG responses to eight different ratios of L- to M-cone contrasts were recorded from each subject. The short-wavelength-sensitive (S-) cone contrast was 0% in all measurements. The recordings were Fourier analyzed to determine the amplitude of the fundamental component. ERG threshold values for each subject resulted in ellipses when plotted in an L-/M-cone contrast space. As expected, the orientations of the threshold ellipses of the protanopes (N = 2) were parallel to the L-cone axis, whereas those of the deuteranopes (N = 2) were parallel to the M-cone axis. For the trichromats (N = 5), there was considerable interindividual variation in ellipse orientation.

Adult↗
PubMed1998

Response phase of the flicker electroretinogram (ERG) is influenced by cone excitation strength.

We measured electroretinogram (ERG) response phases at different cone contrasts in trichromats and dichromats to investigate the dynamics of the long-wavelength-sensitive (L-) and middle-wavelength-sensitive (M-) cone pathways. ERG responses to stimuli, temporally modulated at 30 Hz, were recorded. The stimuli were generated on a computer controlled colour monitor. Thirty-two different combinations of L- and M-cone excitation strength, expressed as cone contrasts, were presented. The short-wavelength-sensitive (S-) cones were not stimulated (S-cone contrast = 0%). The response phase of the fundamental stimulus component was obtained from Fourier analysis. The ERG response phase lags decreased with increasing cone contrast. This was observed in all subjects with a normal appearing fundus. In dichromats and trichromats at low and intermediate contrasts, the phase lags to M-cone isolating conditions were smaller than those to L-cone isolating stimuli. In one dichromat with extreme myopia and cupping of the optic disc, the ERG phase lags increased with increasing cone contrast. The ERG response phase may be potentially useful for detecting retinal abnormalities.

Color Vision Defects↗
PubMed1998

Hue memory and discrimination in young children.

As first remarked by Charles Darwin (1877), very young children frequently have difficulty when naming or choosing colors. To investigate the cause of this difficulty, we have tested preschoolers (mean age = 4.1) for hue discrimination and hue memory and compared their results with those of preadolescents (mean age = 9.6) and young adults (mean age = 25.8). The tests were designed to minimize the influence of verbal coding on the results. We find that preschoolers are as good as the two older groups in hue discrimination. However, in visual hue memory, they are significantly poorer. The 3-fold increased errors they make, relative to preadolescents and young adults, may be related to the development of visual hue categories and the integration of verbal and visual processes. However, such errors cannot explain why young children often experience extreme difficulty in color naming.

Adolescent↗
PubMed1998

Human cone spectral sensitivities: a progress report.

The spectral sensitivities of the short (S-), middle (M-) and long (L-) wave-sensitive cones have been measured in normal trichromats and in dichromats and monochromats of known genotype. For the S-cone sensitivities, three blue-cone monochromats and five normals were used; for the M-cone sensitivities, nine protanopes (three with a single L1M2 gene, three with a single L2M3 gene, one with both an L1M2 and an M gene, and two with both an L2M3 and an M gene); and for the L-cone sensitivities, 22 deuteranopes (five with a single L(ala180) gene and 17 with a single L(ser180) gene). We compare existing cone spectral sensitivity estimates with these results and with tritanopic color matches. The new findings are more consistent with the cone fundamentals of Stockman et al. (JOSA 1993(A10), 2491) than with those of Smith and Pokorny (Vision Research 1975(15), 161). The discrepancies that we find, however, are sufficient to warrant the replacement of both sets.

Color Perception↗
PubMed1998

Numbers and ratios of X-chromosomal-linked opsin genes.

Quantitative Southern blotting and PCR/RFLP analysis were used to determine the number and ratio of long-wave-sensitive (L-) and mid-wave-sensitive (M-) opsin genes in 25 colour-normal caucasian males. The average observed ratio was 1:2.8 +/- 1.2 for Southern blot analysis and 1:3.0 +/- 1.7 for PCR/RFLP analysis. Thus, the two techniques yielded similar results for the ratio of L- to M-opsin genes (Wilcoxon t-test, P < 0.01). PCR/RFLP analysis of a Sma I polymorphism specific for the most proximal opsin gene suggested an average gene number of 6.0 +/- 2.1, with a range from 4 to 12 in individual subjects. In contrast, Southern blot analysis suggested an average number of 3.8 +/- 1.2, with a range from 2 to 7 (on the assumption that only one L-opsin gene is ever present). Differences between the L- to M-opsin gene ratio and the total gene number in some subjects may result from the presence of multiple L-opsin genes and/or hybrid opsin genes in colour-normal males. An exact determination of the total gene number will require employing other molecular techniques.

Blotting, Southern↗
PubMed1998

Total colourblindness is caused by mutations in the gene encoding the alpha-subunit of the cone photoreceptor cGMP-gated cation channel.

Total colourblindness (OMIM 216900), also referred to as rod monochromacy (RM) or complete achromatopsia, is a rare, autosomal recessive inherited and congenital disorder characterized by photophobia, reduced visual acuity, nystagmus and the complete inability to discriminate between colours. Electroretinographic recordings show that in RM, rod photoreceptor function is normal, whereas cone photoreceptor responses are absent. The locus for RM has been mapped to chromosome 2q11 (ref. 2), however the gene underlying RM has not yet been identified. Recently, a suitable candidate gene, CNGA3, encoding the alpha-subunit of the cone photoreceptor cGMP-gated cation channel, a key component of the phototransduction pathway, has been cloned and assigned to human chromosome 2q11 (refs 3,4). We report the identification of missense mutations in CNGA3 in five families with RM. Homozygous mutations are present in two families, whereas the remaining families show compound heterozygous mutations. In all cases, the segregation pattern of the mutations is consistent with the autosomal recessive inheritance of the disease and all mutations affect amino acids that are highly conserved among cyclic nucleotide gated channels (CNG) in various species. This is the first report of a colour vision disorder caused by defects other than mutations in the cone pigment genes, and implies at least in this instance a common genetic basis for phototransduction in the three different cone photoreceptors of the human retina.

Base Sequence↗
PubMed1994

Spatial relations of flicker signals in the two rod pathways in man.

1. Flicker signals originating from the human rod photoreceptors seem to have access to two retinal pathways: one slow and sensitive, the other fast and insensitive. The phase lag between signals in the two pathways grows monotonically with frequency, reaching 180 deg near 15 Hz. 2. At 15 Hz, destructive interference between the slow and the fast signals can cause two related phenomena: (i) a suprathreshold intensity region--the perceptual null--within which the perception of flicker vanishes, and (ii) a double branching of the 15 Hz rod-detected flicker threshold versus intensity (TVI) curve. 3. Here we investigate the effect of changing target size on these phenomena in normal human observers. We find that the double-branched flicker TVI curve and the perceptual null are found for all targets larger than 2 deg in diameter. For smaller diameter targets, however, neither the lower branch of the double-branched flicker TVI curve nor the null are found. 4. While this might suggest that the slow rod signals are selectively disadvantaged by the use of small targets, phase measurements relative to a cone standard reveal that the slow signals are always present. For targets < or = 2 deg in diameter, however, they remain below detection threshold because of destructive interference with the fast rod signals. Thus, for small targets, the perceptual null is not absent, but has merged with (and therefore obliterated) the lower branch of the double-branched flicker TVI function. 5. This situation could arise if decreasing the target size causes a parallel reduction in the sensitivities of both pathways, rather than a selective reduction in the sensitivity of either one. Our findings are therefore consistent with a model in which the large-scale spatial organization of the two rod pathways is roughly similar.

Electrophysiology↗
PubMed1994

Dual rod pathways in complete achromatopsia.

The vision in typical complete achromatopsia is mediated only by rods. To provide more information about the physiological basis of this disorder, we investigated in a typical complete achromat the electrophysiological correlate of the psychophysically established 15-Hz-flicker nulling phenomenon. This phenomenon is believed to be the result of destructive interference between two independent rod pathways. We obtained dark-adapted Ganzfeld electroretinogram (ERG) recordings at 15 Hz with increasing stimulus intensities in the scotopic range and compared the results of the typical complete achromat with those of a normal observer. In both observers the 15-Hz ERG signal reached a minimum at retinal illuminances associated with the perceptual null and reversed in phase as the null was traversed. In comparison with the normal observer, however, the null in the achromat was found at higher scotopic retinal illuminances (ca. 0.4 log unit). Furthermore, the loss in amplitude associated with the null was much more evident, the decline in amplitude was steeper, and the phase shift of 180 degrees was accelerated.

Color Vision Defects↗
PubMed1993

The spectral properties of the two rod pathways.

Psychophysical and electroretinographic observations in normal and achromat observers suggest that rod flicker signals have access to at least two retinal pathways: one (pi 0), slow and sensitive, predominating at scotopic luminance levels; the other (pi'0), fast and insensitive, predominating at mesopic ones. We have measured steady-state flicker detection sensitivities on background fields ranging from 430 to 640 nm in normal observers. Our results suggest that cone signals can reduce the sensitivity of pi'0, but have comparatively little effect on pi 0. The pi'0 field sensitivities derived from these measurements have been fitted with linear combinations of the scotopic luminosity function, V' lambda, the M-cone spectral sensitivity function, M lambda, and the L-cone function, L lambda. These fits demonstrate a clear cone influence on pi'0, but they cannot tell us unequivocally whether the influence is from the M-cones, from the L-cones or from both. Accordingly, we made similar measurements in dichromats, who lack one of the two longer wavelength cone types. These measurements revealed an L-cone influence on pi'0 in the deuteranope and an M-cone influence in the protanope. This suggests that both cone types can affect the sensitivity of pi'0. The finding that the steady-state cone signals reduce the sensitivity of pi'0 but have little effect on pi 0 could suggest that pi'0 signals travel through a faster cone pathway (with its own gain control at which both rod and cone signals can reduce rod threshold), while pi 0 signals travel through a separate rod pathway. However, it could simply reflect the fact that pi'0 predominates at higher luminances than pi 0 where the cone excitation level is inevitably greater. To examine the influence of the cones on pi 0 more closely, we: (i) produced transient cone excitation by alternating rod-equated 480 and 679 nm fields; and (ii) extended our steady-state measurements to include deep-red backgrounds of 650 and 680 nm. Both experiments revealed a small, but measurable influence of the cones on pi 0.

Color Vision Defects↗
PubMed1993

Spatial integration and sensitivity changes in the human rod visual system.

1. The factor by which increment threshold rises with increasing background intensity is less if the target is small than if it is large. The difference is usually attributed to a reduction in the area over which visual signals are integrated as the visual system light adapts. Recently, however, it has been argued that the difference in slope may instead be caused by an increase in the gain of the local response function with light adaptation. 2. To test this hypothesis in the rod-driven visual system, we compared monoptic, small and large target increment thresholds, and dichoptic, large target brightness matches, measured as a function of background intensity in a typical, complete achromat, who has no cone vision. 3. The dichoptic brightness matches were made using a large target of a similar intensity to the threshold intensity of the small target. If local intensity is important, the large target brightness matching curve should be more similar to the shallow, small target threshold curve. But, if changes in spatial integration are important, the brightness matching curve should be similar to the steeper, large target threshold curve. 4. The slope of the large (1.85 deg) target increment threshold functions measured with either 200 or 50 ms test flashes were steeper than those of the small (10 min of arc) target functions by 0.10 (on logarithmic co-ordinates) or about 15%. 5. The logarithmic slopes of the dichoptic brightness curves were also slightly steeper than the small target increment functions. This is contrary to the local response (only) hypothesis, which predicts that the brightness curve should have the same slope as the small target function because the luminance of the targets in the two cases is the same. 6. We conclude that there must be a change in spatial integration in the rod visual system during light adaptation, over and above that due to local gain changes.

Adaptation, Ocular↗
PubMed1993

Temporal and spatial summation in the human rod visual system.

1. Absolute and increment thresholds were measured in a retinal region 12 deg temporal from the fovea with 520 nm targets of varying size and duration. Measurements were made under rod-isolation conditions in two normal observers and in a typical, complete achromat observer who has no cone-mediated vision. The purpose of these experiments was to determine how the temporal and spatial summation of rod-mediated vision changes with light adaptation. 2. The absolute threshold and the rise in increment threshold with background intensity depend upon target size and duration, but the psychophysically estimated dark light of the eye (the hypothetical light assumed to be equivalent to photoreceptor noise) does not. 3. The rise in increment threshold for tiny (10 min of arc), brief (10 ms) targets approaches the de Vries-Rose square-root law, varying according to the quantal fluctuations of the background light. The slope of the rod increment threshold versus background intensity (TVI) curves in logarithmic co-ordinates is about 0.56 +/- 0.04 (when cones are not influencing rod field adaptation). For large (6 deg) and long (200 ms) targets, a maximum slope of about 0.77 +/- 0.03 is attained. 4. The steeper slopes of the rod-detected TVI curves for large, long targets implies some reduction in temporal or spatial summation. In fact, the change in summation area is much more critical: under conditions where only the rod system is active the TVI curve slope is independent of target duration, suggesting that temporal summation is practically independent of background intensity. 5. The rise in threshold also depends on the wavelength of the background field in the normal observer but not in the achromat, confirming reports that the field adaptation of the rods is not independent of the quantal absorptions in the cones. The cone influence is most conspicuous on long-wavelength backgrounds and is found for all target sizes and durations, but is greater for large and long targets than for the other conditions.

Adult↗