Vertebrate rod and cone photoreceptors require continuous supply of chromophore for regenerating their visual pigments after photoactivation. Cones, which mediate our daytime vision, demand a particularly rapid supply of 11-cis retinal chromophore in order to maintain their function in bright light. An important contribution to this process is thought to be the chromophore precursor 11-cis retinol, which is supplied to cones from Müller cells in the retina and subsequently oxidized to 11-cis retinal as part of the retina visual cycle. However, the molecular identity of the cis retinol oxidase in cones remains unclear. Here, as a first step in characterizing this enzymatic reaction, we sought to determine the subcellular localization of this activity in salamander red cones. We found that the onset of dark adaptation of isolated salamander red cones was substantially faster when exposing directly their outer vs. their inner segment to 9-cis retinol, an analogue of 11-cis retinol. In contrast, this difference was not observed when treating the outer vs. inner segment with 9-cis retinal, a chromophore analogue which can directly support pigment regeneration. These results suggest, surprisingly, that the cis-retinol oxidation occurs in the outer segments of cone photoreceptors. Confirming this notion, pigment regeneration with exogenously added 9-cis retinol was directly observed in the truncated outer segments of cones, but not in rods. We conclude that the enzymatic machinery required for the oxidation of recycled cis retinol as part of the retina visual cycle is present in the outer segments of cones.

Photostasis is a phenomenon where the photoreceptor outer segment (OS) length and its rhodopsin content vary depending on environmental lighting. When light is reduced for extended periods, it is argued that OS lengthen and its rhodopsin concentration rises to increase photon capture in darker environment. Increases in OS length may occur because the retinal pigment epithelium (RPE) cells reduce OS consumption in prolonged darkness. But sample sizes in assessing changes in OS length have been small, and results highly varied with no statistical analysis ever offered. Further, animals used were often albinos, which have abnormal RPE cells. Here we keep pigmented and albino mice for 21 days in darkness and compare OS length with those in a normal 12:12 light/dark environment. We measured approximately 1300 OS but found no statistically significant difference in their lengths between light and dark groups in either pigmentation phenotype, although there was a small trend in the data favoring OS extension in the dark. Given that earlier studies were undertaken on limited samples with no statistical analysis, our data pose serious questions for the notion of mammalian photostasis in terms of significant OS plasticity.

The aim of this research was to clarify and quantify the demands of the working environment and watch-keeping regime for large commercial ships in relation to dark adaptation. The night lookout task requires the identification of the relatively bright navigational lights of other ships against the dark background of the sky and sea. The probability of detection is determined by the ambient lighting conditions on the bridge and the dark adapted state of vision. Light levels were such that threshold sensitivity (after 15 minutes) was reduced by around 2 log units in comparison to complete darkness. This has implications for the effective range of navigational lights at sea as defined in regulations. The intensity and position of navigation lights on larger vessels is such that the sensitivity of the eye under typical bridge conditions is likely to be sufficient for their visibility to be acceptable. This may not be the case for less well lit small craft.

The effects of dark adaptation on the response properties of ganglion cells have been documented extensively in the cat retina. To pinpoint the different retinal mechanisms that underlie these effects, we studied the response characteristics of cat horizontal (H) cells during prolonged dark adaptation. H-cell responses were recorded intracellularly in the optically intact, in vivo eye. To disentangle rod and cone contributions, sensitivity changes during dark adaptation were tracked with white light and with monochromatic lights that favored either rod or cone excitation. Stable, long-lasting recordings allowed us to measure changes of sensitivity for adaptation periods up to 45 min. Thresholds for white light and 503-nm monochromatic light decreased steadily and in parallel. The maximum increase of sensitivity, after extinguishing a photopic adaptation light, was 1.8 log units only, reached after about 35 min. Sensitivity for 581-nm lights also increased steadily, but at a shallower slope. The steady increase of sensitivity was concomitant with a linear shift in resting membrane potential and with an increase in relative rod contribution to the threshold responses. Even though small-amplitude responses were rod dominated after prolonged dark adaptation, sensitivity to rod signals remained relatively low, compared to sensitivity of cone responses or to the absolute sensitivity of ganglion cells. This suggests that the cone-H-cell pathway plays no role in the dark-adapted cat retina.

In normal observers, sensitivity of cones to rapid sinusoidal flicker decreases by about 0.7 log units as rods progressively dark adapt. However, Arden and Hogg (1985) described a night-vision disorder characterized by normal rod sensitivity but exaggerated suppressive rod-cone interaction (SRCI). We refer to this condition as the exaggerated SRCI syndrome (ESS). The present paper examines the influence of rod-adaptation upon cone-mediated responses to light onset and offset in an observer with ESS. Under all conditions of adaptation examined, sensitivity of cones to rapid-on waveforms is indistinguishable to that of a normal observer tested under identical circumstances; rod sensitivity is also normal. However, the sensitivity of cones to transient decreases in illumination is clearly subnormal under light-adapted conditions. This deficit in cone responsiveness to light offset becomes increasingly subnormal as rods dark adapt and, when completely dark adapted, the ESS observer is nearly blind to 1 Hz rapid-off sawtooth waveforms. These results strongly bolster previous results that suggest that suppressive rod-cone interaction is restricted to the response to transient decreases in illumination.

Previous studies have reported that the surround responses of retinal ganglion cells weaken or disappear upon dark adaptation. The mechanism(s) by which this occurs is largely unknown, although changes in activity of retinal dopaminergic neurons have been implicated. In the light-adapted rabbit retina, the surround ON responses of OFF-center ganglion cells have been shown to be markedly reduced or abolished by a dopamine antagonist. This effect of a dopamine antagonist was recently shown to be reversed by the glycine antagonist strychnine and by compounds that elevate intracellular cAMP levels. The present study was conducted to determine whether strychnine and cAMP-elevating compounds could bring out the surround ON responses in OFF-center ganglion cells that are diminished upon dark adaptation. Extracellular recordings of OFF-center brisk ganglion cells were made from isolated, superfused retinal preparations. During the course of dark adaptation, the surround On responses of many cells decreased markedly.Application in both brisk-transient and brisk-sustained OFF-center ganglion cells. The center OFF responses of these cells, on the other hand, were not enhanced by strychnine. Of the cAMP-elevating compounds tested, 8-(4-chlorophenylthio) cyclic AMP was the most effective in bringing out the surround ON responses in dark-adapted OFF-center ganglion cells. The findings from this study suggest that under dark-adapted conditions glycinergic neurons inhibit the surround component of OFF-center ganglion cells. The release of glycine from these neurons is suggested to be regulated by a cAMP-dependent mechanism.

The mammalian rod bipolar, for which only one class has been identified, has been described as being hyperpolarizing by some investigators and depolarizing by others. We now report the effects of 2-amino-4-phosphonobutyrate (APB), a potent blocker of depolarizing bipolar cells, on visual behavior in the dark-adapted monkey. While in mesopic and photopic conditions only the monkeys' ability to detect incremental stimuli is impaired, under scotopic conditions all light mediated response in the monkey is eliminated. Assuming APB is acting on rod bipolars in the same fashion as it does on cone bipolars, we conclude that the primate rod bipolars all depolarize to light and that the ON and OFF channels are formed by the amacrine cell network.

In a dichromatic teleost species, we determined the intensity of light of various wavelengths required to prevent cone elongation by exposing fish at the time of their normal “dusk” phase to monochromatic light (479, 623, and 660 nm) at eight to ten different intensities for 75 min. The positions of single and double cones were measured in tangential sections and expressed as cone indices. At all wavelengths, the spectral responses of both cone types were virtually identical. Furthermore, the sensitivity of the blocking effect was highest at shorter wavelengths. When comparing the relative quantal sensitivities of myoid elongation for the two cone types to the spectral sensitivities of the three types of Aequidens pulcher photoreceptor, we found the closest match between the action spectrum and the absorption spectrum of the green-sensitive single cones. This may indicate that this cone type is capable of reacting directly to decreasing levels of illumination. On the other hand, the identical sensitivity of both cone types argues for an indirect control mechanism of dark-adaptive cone elongation, possibly via a neural pathway involving the inner retinal layers, complementary to the neural control of light adaptation. Green-sensitive single cones are well suited to trigger this response, since (1) their sensitivity is inferior to that of double cones; (2) waters inhabited by the blue acara transmit best at long wavelengths; and (3) at dusk, long-wavelength radiation dominates over other parts of the spectrum. Therefore, green-sensitive cone threshold will be reached first at dusk.

Foveal dark adaptation in 58 subjects and photopigment regeneration in 60 subjects from 10–78 years of age exhibit parallel slowing of recovery rate with increasing age, with significant correlation of the two functions among individuals. The data are suggestive of an initial slight decline in rate before age 50, followed by a greater decline occuring at different ages in different ages in different individuals. Longitudinal data for one subject from age 40–65 show an increase in pigment regeneration time constant consistent with this idea. Foveal sensitivity and photopigment density both decrease with increasing age and are significantly correlated among individuals, although sensitivity declines more with age than does photopigment density. In contrast to earlier proposals based upon the Rushton-Dowling equation, we found no universal constant of proportionality to relate log relative threshold to photopigment within our population.

Thresholds for detection of light by a dark-adapted test eye were measured while the other, non-test eye was either similarly dark adapted or while it was exposed to an intense red adapting field. An interocular effect that depends on the retinal location of the test was found: compared to the threshold during binocular dark adaptation, sensitivity decreased during contralateral light adaptation when the test was presented to the foveola and up to 4 deg above it; but sensitivity increased when the test was between 7 and 12 deg, showing a reversal at 5 deg.

Dark-adapted rods exert a tonic suppressive influence upon cone-mediated sensitivity to rapid flicker, a phenomenon called suppressive rod-cone interaction (SRC1). However, rod dark adaptation has negligible influence upon cone-mediated thresholds measured with more usual psychophysical procedures. The present study separately examined the influences of rod light and dark adaptation upon cone-mediated sensitivity to transient increases or decreases in illumination using sawtooth flicker with rapid-on (ramp-off) or rapid-off (ramp-on) waveforms. In the parafoveal retina, cones alone were stimulated with flicker by spatially superimposing longand short-wavelength stimuli presented in counterphase and matched in scotopic illuminance. Several different adaptation procedures were used. For higher (>4 Hz) frequencies, sensitivity of cones to both waveforms is nearly identical under any condition of adaptation; sensitivity decreases as rods progressively dark adapt. A considerably different situation exists for slower frequencies (1–4 Hz). Sensitivity of cones to rapid-off flicker is appreciably greater under light-adapted conditions confirming recent observations by Bowen et al. (1989). But as rods progressively dark adapt, sensitivity of cones to rapid-off waveforms decreases considerably while sensitivity to rapid-on waveforms is much less affected; in the totally dark-adapted eye, sensitivity to both waveforms is identical. These results confirm and extend recent physiological observations in amphibian retina (Frumkes & Wu, 1990) suggesting that SRCI specifically involves responses to transient decreases in illumination.

A photon-counting retinal densitometer is described that has been designed optically and electronically for improved sensitivity and reliability. The device allows measurement of visual pigments through the undilated natural pupils of subjects at relatively low levels of measuring lights, and serves also as an adaptometer for direct comparisons between pigment bleaching or regeneration and light or dark adaptation. Instrumental control and data collection are by computer to permit rapid and simple data analysis and comparisons between subjects. The methods by which the sensitivity and reliability have been enhanced are described in detail, and some examples of experimental results are presented.

The aim of this study was to investigate the effects of an increase in the saturation of blood oxygen (SaO2) and/or serum glucose on photoreceptor sensitivity in normal subjects and in patients with diabetes mellitus. We monitored cone and rod sensitivity by recording dark-adaptation curves to both green and red test stimuli while inhaling either air (20% O2 + 80% N2) or 100% oxygen in 12 normal subjects and 12 diabetic patients with no (10) or mild (2) retinopathy. We also repeated the experiment in 10 of the normal subjects under hyperglycemia (mean serum glucose: 161 mg/dl). Results show that in normal subjects the dark-adapted cone sensitivity is improved by an increase in SaO2 or by hyperglycemia. Final rod sensitivity is unchanged during hyperoxia and during hyperglycemia when measured with a green test spot. However the kinetics of dark adaptation are altered during hyperglycemia, and an increase in final sensitivity is observed when measured with the red test spot. Inhalation of oxygen during hyperglycemia in normal subjects reduces cone sensitivity compared to that found during hyperglycemia alone (Pasteur effect). In diabetic subjects the dark-adapted cone threshold is comparable to that found in normal subjects, and sensitivity also increases with an increase in SaO2. The final rod threshold, however, is impaired compared to that of the control group, and rod sensitivity is improved by increasing the SaO2. The results suggest that the metabolism of rods and cones may differ in normal subjects: in cones, the rate of metabolism can be augmented by increasing the available oxygen or glucose, whereas rods appear more insensitive to increased blood oxygen saturation and hyperglycemia. In diabetic subjects, both cone and rod metabolism can be increased by supplemental oxygen, indicative of an early rod deficit. The study lends weight to the hypothesis that dark-adapted rods in diabetics are hypoxic before the onset of retinopathy.

A fast-scanning dichroic microspectrophotometer was used to trace products of rhodopsin photolysis (metarhodopsins I/II/III and later) in structurally intact amphibian rod outer segments (ROSs) and metabolically active rods. The instrument allows the recording of absorbance spectra with a time resolution better than 1 s, and to discriminate between products with similar absorbance spectra that differ with respect to the orientation of their chromophore in the photoreceptor membrane. We demonstrate that metarhodopsin III is in a pH-reversible equilibrium with metarhodopsin II and that the metarhodopsin III chromophore is orientated with respect to the membrane plane even more strictly than the 11-cis retinal in “dark” rhodopsin. This indicates that all-trans retinal in metarhodopsin III is still attached to its native binding site on opsin. The kinetic scheme of the decay of metarhodopsins is presented in which metarhodopsin III lies in a shunt pathway from metarhodopsin II to retinal. Formation of metarhodopsin III was detected at bleaches as low as ≈ 3%, contrary to previous reports that it is not formed at below 10% bleaches. Another product that is spectrally similar to metarhodopsin III, termed P440, appears at later stages of photolysis as the result of the decay of metarhodopsin II and metarhodopsin III. The chromophoric group in P440 is orientated preferentially across the disk membrane. The final product(s) in isolated ROS, where the reduction of retinal to retinol is blocked, consists of a mixture of a free retinal and retinal possibly attached to different binding sites in the membrane. In metabolically active rods the later products are quickly converted to retinol. We conclude that metarhodopsin III represents a specific conformational state of metarhodopsin where the chromophoric binding site is still occupied by all-trans retinal. Hence, the formation and decay of metarhodopsin III may be limiting for the rate of rhodopsin regeneration and photoreceptor dark adaptation.

In the vertebrate retina, vision is initiated and maintained by the photolysis and regeneration, respectively, of light-sensitive pigments in the disk membranes of the photoreceptor outer segments. This cyclical process depends on an exchange of retinoids between the photoreceptors and the retinal pigment epithelium (RPE). There is a great deal of indirect evidence that the transport of retinoids between these cellular compartments is mediated by the interphotoreceptor retinoid-binding protein (IRBP), a large glycoprotein synthesized in the photoreceptors and extruded into the interphotoreceptor matrix (IPM) that fills the subretinal space. Nevertheless, a number of in vitro experiments have demonstrated that an intermembranous transfer of retinoids can occur through an aqueous medium independent of any retinoid-binding protein. This led to the suggestion that IRBP may play the more passive role of an extracellular buffer, serving to prevent the degradation and potentially cytotoxic effects of free retinoids when large amounts are released into the IPM. We have studied the structural and functional properties of transgenic mice in which homologous recombination was used to delete the IRBP gene. Light- and electron-microscopic examination of the retinas of “knockout” (IRBP−/−) mice revealed a significant loss of photoreceptor nuclei, and profound changes in the structure and organization of the receptor outer segments. Consistent with these observations, electroretinographic recordings showed a marked reduction in response amplitude for both rod- and cone-mediated potentials. However, despite the histological and electrophysiological changes, there was no evidence of gross abnormalities in the visual cycle. After bleaching a significant fraction of the available rhodopsin, electroretinogram amplitude and rhodopsin density gradually increased toward their pre-bleach levels, and the rates of recovery were even more rapid than those seen in wild-type (IRBP+/+) mice.

We have recently developed a behavioral assay, based on the escape response of fish to a threatening object, to analyze quantitatively the visual sensitivity of zebrafish. During the course of dark adaptation, we measure the threshold light intensity required to evoke an escape response. Under a normal light–dark (LD) cycle, thresholds for both the cone and rod systems are considerably lower in late afternoon hours than in early morning hours. Over a period of 24 h, zebrafish are most sensitive to visual stimuli prior to light off and least sensitive prior to light on. Under conditions of constant illumination, this rhythm of visual sensitivity persists for several days but is gradually lost. In constant light (LL), the rhythm persists 1–2 days; thereafter, visual thresholds at all times of the day converge at a level similar to thresholds measured in late afternoon hours in control animals. In constant darkness (DD), the rhythm persists at least 5 days; thereafter, it dampens to a level about a half-log unit less sensitive to that measured in the late afternoon hours in control animals. These data suggest that visual sensitivity in zebrafish is regulated by an endogenous circadian clock which functions to decrease the visual sensitivity.