Field anaesthesia of leatherback sea turtles (Dermochelys coriacea).

Ten nesting leatherback sea turtles on Trinidad were anaesthetised for electroretinogram (ERG) measurements, using ketamine and medetomidine, reversed with atipamezole. They weighed 242 to 324 kg and were given initial doses of 3 to 8 mg/kg ketamine and 30 to 80 microg/kg medetomidine administered into an external jugular vein; six of the turtles received supplementary doses of 2.6 to 3.9 mg/kg ketamine combined with 0 to 39 microg/kg medetomidine. The lower doses were used initially to ensure against overdosage and reduce the chances of residual effects after the turtles returned to the water, but successful ergs called for step-wise dose increases to the required level of anaesthesia. Respiratory rate, heart rate, electrocardiogram, cloacal temperature, and venous blood gases were monitored, and blood was collected for plasma biochemistry. At the end of the erg procedure, atipamezole was administered at 150 to 420 microg/kg (five times the dose of medetomidine), half intramuscularly and half intravascularly. The turtles were monitored and prevented from re-entering the water until their behaviour was normal. No apparent mortalities or serious anaesthetic complications occurred. The observed within-season return nesting rate of the anaesthetised turtles was comparable with that of unanaesthetised turtles.

Aging of the chromatic onset visual evoked potential.

PURPOSE: To examine changes in the chromatic onset visual evoked potential (VEP) as a function of aging. METHODS: VEP's were measured in response to chromatic sinusoidal gratings (1.0 and 0.5 cpd), selectively chosen to modulate the L-M channel and S - (L+M) channel and presented in onset-offset mode. Responses to achromatic gratings presented in a reversal mode were also measured. Twenty subjects were tested, ranging in age from 21 to 93 years. RESULTS: Unlike changes observed earlier in life, the general shape of the chromatic onset wave-form changed little with age; however, latencies increased significantly as a function of age. Amplitude changes revealed a decreasing trend that was not statistically significant. There was little change in the achromatic responses with age. CONCLUSIONS: Our results demonstrate a systematic slowing of the chromatic onset VEP with age. The gradual nature of the latency changes and the lack of dramatic and complex wave-form shape changes may allow development of age-based normative data for use in clinical settings.

Long-term maturation of visual pathways.

Previous research in adults has demonstrated the utility of the visual evoked potential (VEP) to measure the integrity of the chromatic and achromatic visual pathways. The VEP has also been shown to be a valuable indicator of maturation of these pathways in infants up to 1 year of age. The present manuscript reports changes in the visual pathways from 2 years to adulthood as measured by the spatio-chromatic VEP. The responses to achromatic reversal stimuli designed to preferentially activate the low spatial-frequency achromatic (luminance) pathways appear adult-like by 1 year of age. The responses to low spatial-frequency isoluminant onset stimuli designed to preferentially activate the chromatic pathway do not appear as they do in the adult until after 12-13 years of age. The shapes of the chromatic VEP waveforms shift from a positive-negative complex to a negative-positive complex. These changes can be modeled by a decrease in the latency of a large negative component between the ages of 1 year and adulthood. The results suggest that for low spatial-frequency stimuli, there are long-term changes in the development of the chromatic pathways that are not observed in the low spatial-frequency achromatic pathways. The changes in the chromatic VEP waveforms with age may be a physiological correlate of reported behavioral changes.

Analysis of red/green color discrimination in subjects with a single X-linked photopigment gene.

Many subjects despite having only a single X-linked pigment gene (single-L/M-gene subjects) are able to make chromatic discriminations by Rayleigh matching, especially when large fields are used. We used a combination of psychophysics (Rayleigh match), electroretinograms (ERG), and molecular genetic techniques to rule out several possible explanations of this phenomenon. Use of rods for chromatic discrimination was unlikely since strong adapting fields were employed and the large-field match results were not consistent with rod participation. A putative mid- to long-wavelength photopigment that escapes detection by current molecular genetic analysis was ruled out by finding only a single L/M photopigment in flicker ERGs from 16 single-L/M-gene subjects. Large-field match results were not consistent with participation of S cones. Amino acid sequence polymorphisms in the S-pigment gene that might have shifted the S cone spectrum towards longer wavelengths were not found on sequencing. The mechanism of chromatic discrimination in the presence of a single photopigment therefore remains unknown. Further possible explanations such as variations in cone pigment density and retinal inhomogeneities are discussed.

Comparison of color and luminance contrast: apples versus oranges?

Using a spatial, forced-choice, matching protocol, we have measured observers' ability to equate the contrasts of sinusoidal gratings which vary along differing directions in a 3-dimensional color space. In a given experiment, the observer obtained a perceptual match between the contrasts of two gratings whose chromaticities or luminances varied along differing chromatic directions which were selected from among five axes: an achromatic luminance axis (lum), an isoluminant axis where only S-cone activation varied (S-axis), an isoluminant axis where L- and M-cone activation varied in a complementary manner (LM-axis), an axis where only L-cone activation varied (L-axis), and an axis where only M-cone activation varied (M-axis). Even though these chromatic axes were chosen to activate independent mechanisms involved in the early stages of spatiochromatic visual processing, and despite the distinctly differing appearance of patterns from variations along differing directions, we find that observers can reliably make such pairwise contrast matches. Furthermore there is reasonable consistency of matching contrasts among observers and the pairwise contrast matches exhibit the properties of homogeneity and transitivity. This observed homogeneity and transitivity allows, for each color direction, the specification of a single scaling factor which relates perceptual contrast to physical contrast.

Transgenic mice expressing a functional human photopigment.

PURPOSE: Changes in retinal photopigments represent a fundamental step in the evolution of visual systems, in that addition of new pigment types or alterations in the spectral absorption properties of existing pigments modify visual capacities and thus open new visual worlds. To provide a tool that would allow direct examination of the changes caused by the presence of novel photopigments, this study was designed to determine whether a gene encoding a human cone photopigment introduced into the mouse genome would be expressed in a cone-specific manner and would support phototransduction. METHODS: Mice transgenic for the human long wavelength-sensitive (L) photopigment were generated by microinjection of fertilized mouse eggs. RNA expression in different tissues was monitored by reverse transcription-polymerase chain reaction analysis. Photopigment protein was localized in retinal cross sections and wholemounts by antibody staining. Light transduction of the cone photopigments was assessed by flicker photometric electroretinography (ERG). RESULTS: The human transgene was expressed specifically in the mouse cones in quantities comparable to those of the mouse middle wavelength-sensitive (M) pigment gene. Immunocytochemical analysis showed that the human L pigment was abundantly synthesized in most mouse cones, was translocated to the outer segments, and caused no detectable cone degeneration. Electroretinographic spectral sensitivity analysis showed that the human L pigment was efficient in eliciting an electrical response. The degree of expression of the transgene in the two founders correlated well with the spectral responsivity of the ERG. CONCLUSIONS: The human L photopigment transduces light efficiently in mouse cones, implying that all protein domains necessary for efficient interaction with intracellular transport and signal transduction machineries in mouse cones have been conserved through evolution. The expression of the human L photopigment gene in both classes of cone of the mouse retina indicates that the transgene did not have the regulatory elements necessary for restricting its expression to mouse M cones or that such elements are not recognized in mouse UV-sensitive cones.

Development of the spatio-chromatic visual evoked potential (VEP): a longitudinal study.

Most prior visual evoked potential (VEP) research on the development of color vision has employed pattern-reversing stimuli that are not optimal for producing chromatic responses. We measured infant VEPs using low spatial frequency, onset-offset stimuli, modulated along the three axes of a cone-based color space (Derrington et al. [J. Physiol 1984;357, 241-265.]). Three color-normal infants were tested in a longitudinal design over the first postnatal year. One red/green color-deficient infant was also tested at 197 days. We found that VEP responses to S-axis (tritan) stimuli have their initial onset later than responses to red/green (L-M) or achromatic stimuli, and that developmental changes in VEP waveforms are more complex and longer lasting for chromatic than for achromatic stimuli. Possible mechanisms underlying these changes are discussed.

Severity of color vision defects: electroretinographic (ERG), molecular and behavioral studies.

Earlier research on phenotype/genotype relationships in color vision has shown imperfect predictability of color matching from the photopigment spectral sensitivities inferred from molecular genetic analysis. We previously observed that not all of the genes of the X-chromosome linked photopigment gene locus are expressed in the retina. Since sequence analysis of DNA does not necessarily reveal which of the genes are expressed into photopigments, we used ERG spectral sensitivities and adaptation measurements to assess expressed photopigment complement. Many deuteranomalous subjects had L, M, and L-M hybrid genes. The ERG results showed that M pigment is not present in measurable quantities in deutan subjects. Using these results to determine gene expression improved the correlations between inferred pigment separation and color matching. Furthermore, we found a subject who had normal L and M genes and normal proximal promoter sequences, yet he had a single photopigment (M) by ERG and tested as a protanope. These results demonstrate the utility of ERG measurements in studies of molecular genetics of color vision deficiencies, and further support the conclusion that not all genes are expressed in color deficient subjects. In particular, deuteranomaly requires a presently unknown mechanism of selective expression which excludes normal M genes and allows expression of L-M hybrid genes in one cone type, and the normal L in another.

Development of pattern visual evoked potentials: longitudinal measurements in human infants.

PURPOSE: This experiment used longitudinal testing to trace the emergence of the major components of pattern visual evoked potentials (VEPs) in infants, using two paradigms: large-checkerboard pattern reversal and low spatial frequency pattern onset. METHODS: Testing with both pattern-reversal and pattern-onset stimuli was performed on the same infants. Testing was conducted at weekly intervals during the first three postnatal months, and at intervals of 2 weeks to 1 month thereafter. RESULTS: The pattern-reversal and early pattern-onset responses recorded within individual subjects showed remarkably systematic developmental sequences. The broad, positive component seen at 200 to 250 ms in infants could be traced readily through the developmental sequence, to become the more sharply tuned positive component seen at about 100 ms in adults. Responses to low spatial frequency pattern onsets in infants were larger and more reliable than those in adults. The late components of the pattern-onset response, generally attributed to pattern offset, emerged later and with more complex changes. In all cases, response amplitude was much more variable than response latency, both within and between subjects. CONCLUSIONS: Frequent VEP recording in a longitudinal design can reveal systematic and detailed transitions of wave-form during development.

Temporal response characteristics of the spatiochromatic visual evoked potential: nonlinearities and departures from psychophysics.

Although the visual evoked potential (VEP) for isoluminant stimuli has been characterized in terms of spatiochromatic parameters, temporal tuning along various chromatic directions has received less systematic attention. Additionally, there has been little categorical comparison of psychophysical appearance with VEP responses obtained for temporal variation of these patterns. At appropriate contrasts the VEP's for color axes (LM, S) show a robust and contrast-sensitive temporal tuning peak at 4 Hz. Contrast response functions at 4 Hz for the LM color axis are markedly nonmonotonic. However, there is a clear monotonicity with contrast for VEP latencies along these color axes. The anomalous behavior does not appear to be due to interactions between chromatic signals, to luminance artifact, or to rod intrusion. These anomalies in the temporal characteristics of the chromatic VEP may reflect interactions between chromatic responses and inherent cortical responsivity not linked to psychophysical behavior.

Contribution of chromatic mechanisms to the production of small-field optokinetic nystagmus (OKN) in normals and strabismics.

The contribution of chromatic mechanisms to motion processing is currently debated. Although OKN as a measure of motion processing has been employed for chromatic stimuli, the contribution of chromatic mechanisms to the production of OKN has not been assessed directly. We measured voluntary-pursuit and involuntary-stare OKN responses to drifting patterns defined by colors modulated in a cone based color space which isolates luminance and color mechanisms. Results show that OKN responses are reduced for isoluminant stimuli, particularly for the stare conditions. The greatest reduction in responses occur for isoluminance stimuli that isolate the S cones. These results suggest that the chromatic mechanisms may have reduced input to the neural substrate that produces involuntary-stare OKN or that stare OKN has different temporal characteristics when stimulated by chromatic and luminance mechanisms. We also measured OKN asymmetries in several convergent strabismics in response to isoluminant and luminance patterns and compared their response to previous reports of horizontal OKN asymmetries in normal neonate infants. The results question the validity of either the modeling of OKN asymmetries in strabismus as incomplete development or the comparison of adult eye movement records evaluated with criteria for OKN with psychophysical forced choice evaluation of ocular drift in infants.

Photopigments and color vision in the nocturnal monkey, Aotus.

The owl monkey (Aotus trivirgatus) is the only nocturnal monkey. The photopigments of Aotus and the relationship between these photopigments and visual discrimination were examined through (1) an analysis of the flicker photometric electroretinogram (ERG), (2) psychophysical tests of visual sensitivity and color vision, and (3) a search for the presence of the photopigment gene necessary for the production of a short-wavelength sensitive (SWS) photopigment. Both electrophysiological and behavioral measurements indicate that in addition to a rod photopigment the retina of this primate contains only one other photopigment type--a cone pigment having a spectral peak ca 543 nm. Earlier results that suggested these monkeys can make crude color discriminations are interpreted as probably resulting from the joint exploitation of signals from rods and cones. Although Aotus has no functional SWS photopigment, hybridization analysis shows that Aotus has a pigment gene that is highly homologous to the human SWS photopigment gene.

Application of the spatiochromatic visual evoked potential to detection of congenital and acquired color-vision deficiencies.

Visual evoked potentials were recorded in response to spatiochromatic stimuli modulated in different directions in cone-activation color space from subjects with congenital and acquired color defects. This technique was effective for detection and classification of both mild and severe forms of congenital deficits. Results suggest that the visual evoked potential is useful for early identification of color abnormalities in acquired deficits such as diabetes and that it is sensitive enough to detect regional retinal losses of sensitivity (e.g., as in central serous choroidopathy). The spatiochromatic visual evoked potential provides a systematic and sensitive indication of different color-vision anomalies.

Photopigments of dogs and foxes and their implications for canid vision.

Electroretinogram (ERG) flicker photometry was used to examine the photopigment complements of representatives of four genera of Canid: domestic dog (Canis familiaris), Island gray fox (Urocyon littoralis), red fox (Vulpes vulpes), and Arctic fox (Alopex lagopus). These four genera share a common cone pigment complement; each has one cone pigment with peak sensitivity of about 555 nm and a second cone pigment with peak at 430-435 nm. These pigment measurements accord well with the conclusions of an earlier investigation of color vision in the dog, and this fact allows some predictions about color vision in the wild canids. An additional set of measurements place the peak of the dog rod pigment at about 508 nm.

Behavioral and electrophysiological sensitivity to temporally modulated visual stimuli in the ground squirrel.

Behavioral and electrophysiological methods were used to measure sensitivity to flickering lights in a dichromatic species, the California ground squirrel (Spermophilus beecheyi). Discrimination tests were used to determine spectral sensitivity at stimulus frequencies from 5-50 Hz and increment threshold spectral sensitivity. The contributions of retinal mechanisms to these capacities were assessed by recording the responses of optic nerve fibers to temporally modulated monochromatic lights. In the ground squirrel, as in the human, the shape of the spectral-sensitivity function depends on the temporal frequency of the stimulus. Results from single-unit recording show that all of the classes of optic nerve fibers in the ground squirrel are highly phase-locked to the stimulus for modulation rates as high as 50 Hz. Neither the responses of photoreceptors nor any class of optic nerve fiber can singly account for the behavioral results. The electrophysiological results are also counter to models which propose that temporally dependent changes in the spectral sensitivity of spectrally opponent fibers account for the behavior. The temporal resolution of the optic nerve fibers exceeds that of the behaving animal suggesting that retinal mechanisms do not limit behavioral temporal resolution.