Green-sensitive Cones Research Articles

Green-sensitive cone photoreceptors are selectively labeled by Procion yellow dye in goldfish retina

Selective labeling of intravitreal Procion yellow dye by presumed blue-sensitive cone photoreceptors has been demonstrated in primate retina. To determine whether Procion yellow is selective for this cone type in an unrelated vertebrate species, labeling by this dye was studied in goldfish retina, where cone pigment type can be directly inferred from photoreceptor morphology. At low vitreal concentrations of the dye (less than 0.4%), only cone outer segments were labeled. At vitreal concentrations of 0.4-0.5%, the inner segments of short-double cones and a subset of long single cones (presumed green-sensitive cones) were selectively stained. At still higher vitreal concentrations (0.6-0.7%), the inner segments of short-single cones and miniature short-single cones (presumed blue-sensitive cones) showed evidence of Procion label, but were not as heavily labeled. The inner segments of long-double cones and a subset of long-single cones (presumed red-sensitive cones) did not label at any of these concentrations. These results show that Procion yellow is not a selective marker for blue-sensitive cones in the goldfish retina. In addition, stained rod and cone nuclei were observed at each dye concentration, including those concentrations at which no inner segments were labeled.

Patterns of glutamate immunoreactivity in the goldfish retina

Postembedding silver-intensified immunogold procedures reveal high levels of glutamate immunoreactivity in "vertical" elements of the goldfish retina: (1) Red-sensitive and green-sensitive cones display strong glutamate immunoreactivity, especially in their synaptic terminals, but blue-sensitive cones are poorly immunoreactive. (2) All type Mb (on-center) and Ma (off-center) mixed rod-cone bipolar cells and all identifiable cone bipolar cells are highly glutamate immunoreactive. We find no evidence for bipolar cells that lack glutamate immunoreactivity. (3) The majority of the somas in the ganglion cell layer and certain large cells of the amacrine cell layer resembling displaced ganglion cells are strongly glutamate immunoreactive. (4) Despite their high affinity symport of acidic amino acids, the endogenous levels of glutamate in Müller's cells are among the lowest in the retina. (5) GABAergic neurons possess intermediate levels of glutamate immunoreactivity. Quantitative immunocytochemistry coupled with digital image analysis allows estimates of intracellular glutamate levels. Photoreceptors and bipolar and ganglion cells contain from 1 to 10 mM glutamate. The bipolar and ganglion cell populations maintain high intracellular glutamate concentrations, averaging about 5 mM, whereas red-sensitive and green-sensitive cones apparently maintain lower levels. Importantly, photoreceptor glutamate levels are extremely volatile, and in vitro maintenance is required to preserve cone glutamate immunoreactivity in the goldfish. GABAergic horizontal and amacrine cells contain about 0.3-0.7 mM glutamate, which matches the values predicted from the Km of glutamic acid decarboxylase. Müller's cells and non-GABAergic amacrine cells contain less than 0.1 mM glutamate. Though Müller's cells are known to possess potent glutamate symport, they clearly possess equally potent mechanisms for maintaining low intracellular glutamate concentrations.

Visual transduction in cones of the monkey Macaca fascicularis.

1. Visual transduction in macaque cones was studied by measuring the membrane current of single outer segments projecting from small pieces of retina. 2. The response to a brief flash of light was diphasic and resembled the output of a bandpass filter with a peak frequency near 5 Hz. After the initial reduction in dark current there was a rebound increase which resulted from an increase in the number of open light-sensitive channels. The response to a step of light consisted of a prominent initial peak followed by a steady phase of smaller amplitude. 3. Responses to dim light were linear and time-invariant, suggesting that responses to single photons were linearly additive. From the flash sensitivity and the effective collecting area the peak amplitude of the single photon response was estimated as about 30 fA. 4. With flashes of increasing strength the photocurrent amplitude usually saturated along a curve that was gentler than an exponential but steeper than a Michaelis relation. The response reached the half-saturating amplitude at roughly 650 photoisomerizations. 5. The response-intensity relation was flatter in the steady state than shortly after a light step was turned on, indicating that bright light desensitized the transduction with a delay. This desensitization was not due to a reduction in pigment content. In the steady state, a background of intensity I lowered the sensitivity to a weak incremental test flash by a factor 1/(1 + I/IO), where IO was about 2.6 x 10(4) photoisomerizations s-1, or about 3.3 log trolands for the red- and green-sensitive cones. 6. Bleaching exposures produced permanent reductions in flash sensitivity but had little effect on the kinetics or saturating amplitude of subsequent flash responses. The sensitivity reductions were consistent with the expected reductions in visual pigment content and gave photosensitivities of about 8 x 10(-9) microns2 (free solution value) for the red- and green-sensitive pigments. During a steady bleaching exposure the final exponential decline of the photocurrent had a rate constant given by the product of the light intensity and the photosensitivity. 7. In some cells it was possible to measure a light-induced increase in current noise. The power spectrum of the noise resembled the spectrum of the dim flash response and the magnitude of the noise was consistent with a single photon response roughly 20 fA in size. 8. The membrane current recorded in darkness was noisy, with a variance near 0.12 pA2 in the band 0-20 Hz.(ABSTRACT TRUNCATED AT 400 WORDS)

Wavelength-dependent temporal properties of retinal horizontal cells in turtles

Electrical responses of luminosity horizontal cells (L cells) to monochromatic stimuli were analyzed by intracellular recordings in the retinas of the freshwater turtle (Pseudemys scripta elegans) and of the sea turtle (Chelonia mydas mydas). Light intensity, duration, and wavelength were varied to assess temporal effects. For a given intensity of monochromatic light, response amplitude increased with stimulus duration until maximum amplitude occurred at a specific duration. This suprathreshold metric of temporal integration is called here summation time, and it is wavelength-dependent. L cells always had some level of red-sensitive cone input, although in some cells inputs from green- and blue-sensitive cones were also observed. For these latter cells, summation times were shorter for 640-nm than for 540-nm or 450-nm lights. These results were most evident in cells that received dominant inputs from blue- or green-sensitive cones. Responses of some other L cells were almost completely dominated by inputs from red-sensitive cones. Summation times of these cells were not wavelength-dependent. But when these inputs also included green-sensitive cones, shorter summation times were obtained to 640-nm light than to 540-nm light, even though dominant inputs were still from red-sensitive cones. These results, obtained from both retinal and 3,4-dehydroretinal photopigment systems, are consistent with reported observations in Pseudemys scripta elegans that show linear responses of red-sensitive cones to have shorter integration times and times-to-peak than green-sensitive cones. Responses from horizontal cells dominated by blue-sensitive cone inputs were the most sensitive of all; they also had the longest summation times.(ABSTRACT TRUNCATED AT 250 WORDS)

Correlation of spinule dynamics and plasticity of the horizontal cell spectral response in cyprinid fish retina: quantitative analysis

A negative feedback interaction between luminosity type horizonatal cells (HCs) and green-sensitive cones generates the long-wavelength-sensitive depolarizing response in biphasic chromaticity type HCs. This interaction is suppressed in the dark and is potentiated by light adaptation of the retina. HCs are morphologically ‘plastic’; during light adaptation, their dendritic terminals within cone pedicles extend, giving rise to “spinules”. This paper examines whether there is a quantitative correlation between the time course of light-dependent formation of the spinules and enhancement of the feedback interaction. The strength of the feedback interaction in isolated retinac of the roach was determined as the ‘neutral wavelength’ at which reversal of spectral response polarity occurred in biphasic HCs. A good correlation was found between the ‘neutral wavelength’ and the spinule/ribbon ratios of retinae. Biphasic HCs were intracellularly stained with horseradish peroxidase and the correlative ultrastructure of the contacted pedicles was examined. ‘Neutral wavelength’ was found to be correlated with the spinule number, weighted according to the number of synaptic contacts mediating feed-forward transmission. The latter was estimated from the total number of labelled Cb/H2 HC processes (central and lateral) at synaptic triads. A model in which spinules mediate the negative feedback interaction of HCs in the retina of cyprinid fish is presented.

Telodendrial contact of HRP‐filled photoreceptors in the turtle retina: Pathways of photoreceptor coupling

Synaptic contacts of photoreceptors in the turtle retina were studied by intracellular injection of horseradish peroxidase (HRP) and electron microscopy. Both cone and rod photoreceptors radiated basal processes (telodendria) from their terminal endings. These telodendria ran laterally in the outer plexiform layer. The telodendria of cones gave rise to many fine branches that penetrated synaptic cavities of several neighboring cones. Tips of these branches terminated near the walls of synaptic cavities. Some of the telodendrial contact formed two types of basal junction: symmetrical and punctate. The distribution of cones that made telodendrial contacts with the HRP-filled cone were quantitatively investigated. Green-sensitive cones (n = 3) made telodendrial contacts with neighboring red- and blue-sensitive cones, blue-sensitive cones (n = 4) with red- and green-sensitive cones, and red-sensitive cones (n = 9) with red- and green-sensitive cones. In contrast to these cone connections, rod telodendria did not penetrate neighboring photoreceptors. Direct synaptic contacts were not found between rods and cones. Our results clarify the variety of cone couplings in turtle retina: the three chromatic classes of cones are selectively coupled by the basal junctions at the ends of telodendrial processes.

Quantitative analysis of cone photoreceptor-horizontal cell connectivity patterns in the retina of a cyprinid fish: electron microscopy of functionally identified and HRP-labelled horizontal cells.

Horizontal cells generating photopic luminosity and biphasic/chromaticity-type S-potentials were identified and intracellularly labelled with horseradish peroxidase in the retina of the roach. The synaptic connectivity patterns of the horizontal cell dendrites within pedicles of different spectral types of cone were then quantitatively studied by electron microscopy. Luminosity-type responses were generated by H1-like horizontal cells contacting similar numbers of red- and green-sensitive cones and very few blue-sensitive cones. Most dendritic contacts were lateral to synaptic ribbons. Central contacts with ribbons were made almost exclusively within red-sensitive cone pedicles. Biphasic/chromaticity-type S-potentials were generated by H2-like horizontal cells. The dendrites of the latter contacted green- and blue-sensitive cones, both at central and lateral sites at synaptic ribbons. An attempt was made to correlate cone ribbon connectivity patterns and spectral characteristics of the horizontal cells according to several hypotheses, some proposed in earlier studies.

Carbohydrate components recognized by the cone-specific monoclonal antibody CSA-1 and by peanut agglutinin are associated with red and green-sensitive cone photoreceptors.

Previous investigations have demonstrated that the monoclonal antibody CSA-1 and peanut agglutinin label specifically cone photoreceptor cells. In the present study, we compared the binding of CSA-1 and peanut agglutinin to that of the monoclonal antibodies COS-1 and OS-2, which have been shown to recognize the red/green- and blue-sensitive cone visual pigments, respectively. Using lectin and immunocytochemistry on serial semithin sections of the pig retina, we have demonstrated in the present study that both CSA-1 and peanut agglutinin label specifically the red-, and green-sensitive, but not the blue-sensitive cone cell outer segments. Peanut agglutinin does bind, however, to the cone matrix sheaths associated with all three types of cones. These observations support the idea that red-, and green-sensitive cone cells share some common molecular epitopes and may represent a differentiation line of cones, considerably different from that of blue-sensitive cones.

Lateral feedback from monophasic horizontal cells to cones in carp retina. II. A quantitative model.

About half of the monophasic horizontal cells in carp retina receive input from both red- and green-sensitive cones. Since the horizontal cells feed back to cones, the color and feedback pathways result in wavelength- and intensity-dependent changes of the dynamics and of the receptive field amplitude profile of the horizontal cell responses. In this paper we present a quantitative model that describes adequately the color and spatial coding and the dynamics of the responses from monophasic horizontal cells in carp. Lateral feedback plays a distinct role in this model.

Rods trigger light adaptive retinomotor movements in all spectral cone types of a teleost fish

Action spectra for light induced cone contraction are described for the two spectral cone types of a dichromatic (red/green) cichlid species ( Aequidens pulcher). Criterion response thresholds (50% of maximal response amplitude) were determined for seven wavelengths. After correcting for absorption by cornea and lens, the resulting action spectra were compared with the absorption spectra of the rod and cone visual pigments. We find (i) that the action spectra of red and green sensitive cones are almost identical and (ii) matched most closely the absorption spectrum of the rod visual pigment. We therefore conclude that light adaptive cone contraction is triggered by light absorption in rods, which in the dark adapted state are located next to the external limiting membrane and are therefore in an optimal position to capture the incident light during early dawn. Possible mechanisms of signal transfer from rods to cones are discussed.

Psychophysically derived visual mechanisms in turtle. II—Spatial properties

Visual mechanisms isolated in Pseudemys by the two-color threshold technique of Stiles show peak wavelength sensitivities at 650 nm (red light) and 540 nm (green light). Ricco critical areas were measured for the two test wavelengths under three conditions: dark, moderate and intense backgrounds. As expected, critical spatial areas decreased with light adaptation. Under dark adaptation only rods and red-sensitive cones were operative, and one photon per 12 rods was sufficient for green-light threshold, as was one photon per four red-sensitive cones for red-light threshold. Rods apparently pool their information among the several receptors within the threshold area. Under light adaptation, rods were not functional and thresholds were determined by red-sensitive and green-sensitive cones alone. Cones did not share information over many receptors, requiring close to one photon per receptor to function at threshold.

H1 horizontal cells of carp retina have different postsynaptic mechanisms to mediate short- versus long-wavelength visual signals.

Vertebrate photoreceptors release neurotransmitter substance(s) tonically in the dark and this release is curtailed by light. Recently, we have become increasingly aware of the possibility that short- and long-wavelength visual signals are mediated differently during the synaptic transmission to second-order retinal neurons. The experiment described here advances this notion further by demonstrating a postsynaptic difference. Treatment of the carp retina by dopamine reduced the gap-junctional coupling of horizontal cells, and we made use of this known effect to measure the input resistance (Rin) of H1-type horizontal cells. Flashes of light increased Rin. This increase, however, was found to be smaller with short wavelengths, even though the comparison was made when voltage responses were equal in amplitude. Often, Rin was even found to decrease at the blue end of spectrum. No single postsynaptic mechanism can account for any equal-voltage Rin difference such as this. The synaptic spectral segregation thus revealed is probably subserved by a dual scheme wherein the transmitter from blue-sensitive cone photoreceptors acts to decrease the membrane conductance of H1 cells whereas the synapses made by red- and green-sensitive cones are of a classical excitatory type.

Psychophysically derived visual mechanisms in turtle. I—Spectral properties

Two-color increment thresholds were measured in turtle using a behavioral avoidance paradigm. Four visual mechanisms were isolated, and their spectral sensitivities were compared to action spectra of photoreceptors known to occur in this animal. One visual mechanism appears to be mediated by rods and red-sensitive cones at low background intensities, a second mechanism by coupled red- and green-sensitive cones at intermediate levels, a third and fourth mechanism by red-sensitive, and green-sensitive single cones, respectively, acting alone at high background intensities.

A horizontal cell selectively contacts blue-sensitive cones in cyprinid fish retina : intracellular staining with horseradish peroxidase

A horizontal cell selectively contacting blue-sensitive cones has been intracellularly stained with horseradish peroxidase in the retina of a cyprinid fish, the roach. The light microscopical morphology of the cell belonged to the H3 category of horizontal cells found in cyprinid fish retinae. In response to spectral stimuli, the cell generated chromaticity-type S-potentials that were hyperpolarizing to blue and depolarizing to yellow-orange. A red-sensitive hyperpolarizing component was absent possibly because of suppression of the negative feedback pathway between luminosity-type (H1) horizontal cells and green-sensitive cones.

Plasticity of cone horizontal cell functioning in cyprinid fish retina: effects of background illumination of moderate intensity.

In electrophysiological experiments involving intracellular recording from horizontal cells in the isolated retina of the roach, light adaptation of the retina has been shown to result in potentiation both of (1) the depolarizing component of biphasic chromaticity type S-potentials, and (2) the temporal frequency transfer functions of photopic luminosity type horizontal cells. Under identical light adaptation conditions, the number of spinules on horizontal cell dendrites positioned laterally at cone pedicle ribbon synapses, increase by some threefold. The latter effect occurs equally in pedicles of red- and green-sensitive cones. Thus, horizontal cells are 'plastic' in both structural and electrophysiological respects. Furthermore, since the two electrophysiological parameters studied depend on negative feedback from horizontal cells onto cones, the results suggest that it is the inhibitory synapses that are plastic and that spinules may be sites of the negative feedback interaction. Physiological and behavioural aspects of light-dependent horizontal cell plasticity are also discussed.

Photocurrents of cone photoreceptors of the golden-mantled ground squirrel.

1. Visual transduction in photoreceptors of the ground squirrel, Citellus lateralis, was studied by recording membrane current from individual cones in small pieces of retina. 2. Brief flashes of light produced transient reductions of the dark current; saturating response amplitudes were up to 67 pA. A flash strength of about 11,000 photons microns-2 at lambda max was required to give a half-saturating response. The stimulus-response relation was well fitted by an exponential saturation curve. Responses below 20% of maximum behaved linearly. 3. The response to a dim flash in most cells had a time to peak of 20-30 ms and resembled the impulse response of a series of five low-pass filters. 4. The variance of the dim-flash response amplitude put an upper limit of 80 fA on the size of the single photon response. Estimates based on the effective collecting area suggest the single photon response to be of the order of 10 fA. 5. Flash responses of squirrel cones usually lacked the undershoot observed in primate cones, although in about 1/3 of the cells a small undershoot developed during recording. 6. Background lights slightly shortened the time to peak of the flash response and reduced the integration time. 7. Spectral sensitivity measurements showed two classes of cones with peak sensitivities at about 520 and 435 nm. Rod sensitivity peaked near 500 nm. Spectral univariance was obeyed by all three classes of cells. 8. The shapes of the spectral sensitivity curves of the rod and both types of cones were similar to each other when plotted on a log wave number scale, but differed significantly from similar plots of monkey and human cone spectra. 9. The kinetics and sensitivity of flash responses of the blue- and green-sensitive cones were indistinguishable.

FOVEAL AND NON‐FOVEAL MEASURES OF SHORT WAVELENGTH SENSITIVE PATHWAYS IN GLAUCOMA AND OCULAR HYPERTENSION*

Blue-sensitive cone thresholds (the ability to detect blue test lights presented on a bright yellow background) were measured to examine foveal and central vision field sensitivity in groups of 33 glaucoma, 31 ocular hypertension (OHT) and 24 age matched normal subjects. Results showed these thresholds were substantially reduced (in all areas tested) for the glaucoma group (approximately 0.8 log units) and less reduced for the OHT group (approximately 0.2 log units), when compared to the normal group threshold. Measures of red-sensitive and green-sensitive cone thresholds showed no difference for all three groups. Furthermore, 16% of the OHT group showed visual field abnormalities for the blue-sensitive cone threshold measure. We conclude that efforts to detect early function loss in glaucoma and OHT could be improved using these measures in clinical practice.

Identification of new biphasic C-type horizontal cells in cyprinid fish.

New biphasic C-type horizontal cells were identified physiologically and morphologically in the retina of live immobilized crucian carp. These cells, like commonly studied R/G type cells, hyperpolarized to short wavelength lights and depolarized to long wavelength lights, but inverted response polarity at about 560 nm. Spectral sensitivities of hyperpolarizing and depolarizing components showed that they were produced by inputs from blue-sensitive cones and green-sensitive cones respectively. These cells are thus referred to as G/B type horizontal cells. Inputs from blue- and green-sensitive cones were not linearly summated, but interacted strongly at G/B type cells. G/B type cells stained intracellularly with HRP had smaller cell body and dendritic field, distinctly differing from R/G type cells.

Photoreceptor inputs converging onto C-type horizontal cells of the retina of crucian carp.

Responses of R/G-type horizontal cells recorded intracellularly in eye cup preparations with normal circulation of crucian carp under dark-adapted conditions were studied and photoreceptor inputs converging onto them were identified. Spectral sensitivities of the hyperpolarizing component of responses, determined at different phases, suggested that it was produced by input from green-sensitive cones. This input and input from long wavelength sensitive cones, causing depolarization of cells, were almost linearly summated. Based on the linear summation assumption, an algebraic procedure was developed to isolate the depolarizing component from responses of R/G-type cells to flashes at wavelengths longer than 540 nm. Spectral sensitivities of the isolated depolarizing component were determined and agreed reasonably well with those of red-sensitive cones. Therefore, it is unlikely that depolarization of R/G-type cells may be due to input from some presumptive deep-red sensitive cones.

Foveal and non-foveal measures of short wavelength sensitive pathways in glaucoma and ocular hypertension

Abstract— Blue-sensitive cone thresholds (the ability to detect blue test lights presented on a bright yellow background) were measured to examine foveal and central vision field sensitivity in groups of 33 glaucoma, 31 ocular hypertension (OHT) and 24 age matched normal subjects. Results showed these thresholds were substantially reduced (in all areas tested) for the glaucoma group (˜ 0.8 log units) and less reduced for the OHT group (˜0.2 log units), when compared to the normal group threshold. Measures of red-sensitive and green-sensitive cone thresholds showed no difference for all three groups. Furthermore, 16% of the OHT group showed visual field abnormalities for the blue-sensitive cone threshold measure. We conclude that efforts to detect early function loss in glaucoma and OHT could be improved using these measures in clinical practice.