Suppression of cone signal in the dark-adapted frog retina as indicated by the electroretinogram.

Electroretinogram (ERG) cone acitvity is depressed in the dark-adapted frog retina. The strength of this effect is examined over a large range of flash energy, for 618 nm flashes extending up to about 4 log10 units above the "threshold" (10-25 micron V b-wave) of the cone ERG "released" in the early stage of rapid dark-adaptation (RDA). Cone signal depression is remarkably strong over this flash energy range. The cone ERG is practically absent for flashes up to about I log unit above cone RDA threshold. For stronger flashes, the suppression becomes time-dependent, that is, cone signal is very small for the first few hundred msec. after the flash, cone intrusion then becoming detectable. The results suggest that the cone suppression phenomena arises distally in the retina, probably near the receptor layer, and that cone signal intrusion a few hundred msec. after a strong flash may be due to light-adaptation of rods by the flash itself.     

Activity of long-wavelength cones under scotopic conditions in the cyprinid fish Danio aequipinnatus

In carp (Cyprinus) and goldfish (Carassius), long-wavelength cones are reported to be active under scotopic conditions. Using the electroretinogram (ERG), we tested another cyprinid fish, Danio aequipinnatus, which contains A₁-based visual pigments and for which we had previously measured the spectral sensitivities of individual cones. Dark adaptation curves show a rod/cone break at about 45 min. When thoroughly dark-adapted, the spectral sensitivity function is broader than can be accounted for by self-screening of rhodopsin, but it can be modeled by an additive combination of rods and the 560-nm cones. Dim, red background light causes adaptation of rods and a broadening of the spectral sensitivity function, which can be simulated by increasing the proportion of cones in the model. Brighter red backgrounds adapt the 560-nm cones. Because of the effect of red adapting lights, the ERG evidence for the participation of long-wavelength cones close to visual threshold appears to be different in Danio than in the goldfish Carassius. Accepted: 14 June 1997     

Electrophysiological evidence for rod and cone-based vision in the nocturnal flying squirrel

Summary The flying squirrel (Glaucomys volans) is a strongly nocturnal rodent. Previous anatomical observations suggested that the retina of this animal contains some cone-like receptors in addition to large numbers of rods. Evidence for duplicity of function in this visual system was obtained from an examination of three indices of visual activity: the electroretinogram (ERG), the isolated PIII retinal response, and the visually evoked cortical potential (VECP). The spectral sensitivity of the dark-adapted flying squirrel is similar to that of other mammals — it has a 500 nm peak (Figs. 3, 8). Responses of the ERG and isolated PIII to flickering light indicate the operation of two processes (Figs. 4, 7), one of which is unable to follow flickering light at repetition rates above 10–15 Hz. Spectral sensitivity measurements reveal that these two processes have different spectral sensitivities. The photopic mechanism in the flying squirrel visual system has peak sensitivity at about 520 nm (Figs. 5, 7, 9). The effects of steady light adaptation are much more obvious in the cortical potentials than they are in the retinal potentials.We thank David Birch for his advice and assistance. This research was supported by a Grant from the National Eye Institute (EY-00105).     

Spectral mechanisms in the tree squirrel retina

Summary The retina of the gray squirrel (Sciurus carolinensis) contains rods and cones in a ratio of about 23. The spectral mechanisms in this retina were examined in behavioral and electrophysiological experiments. Tests of color vision revealed that this animal has a spectral neutral point at about 500 nm and, thus, dichromatic color vision. Recordings made from single optic nerve fibers and results obtained from an analysis of the flicker photometric electroretinogram (ERG) indicated that vision in the gray squirrel is based on three spectral mechanisms. One of these, presumably rod-based, has peak sensitivity at about 502 nm. The other two mechanisms reflect the presence of two classes of cone having average peak sensitivity of about 444 nm and 543 nm.     

Spectral sensitivity of the cod,Gadus morhua L.

The spectral sensitivity of the cod was determined under both dark adapted and light adapted conditions in the laboratory. Cod were trained by cardiac conditioning to detect a difference in radiance between an image of spots and the background radiance of a screen. Thresholds for this response were measured for a range of different wavelengths, and expressed as quantum adjusted values. Electroretino‐graphic studies were also performed on the eyes of cod, and spectral sensitivity curves prepared. Under dark adapted conditions both the behavioural and e.r.g. derived curves showed greatest sensitivity in the blue/green at 490 nm, matching the absorption curve for rhodopsin. A secondary peak in the behaviourally derived curve in the green/yellow at 550 nm indicated that a population of yellow cones may be implicated with the rods in scotopic vision. Under light adapted conditions the behavioural curves showed a shift to the blue, perhaps indicating an adaption to the high red content of the illuminating source. The e.r.g. curve showed greatest sensitivity to blue/green, as in the scotopic experiments but with an enhanced response at 550 nm, indicating greater cone activity. It is suggested that there is complex interaction between rods and cones in the cod retina, both types of receptor being active over a wide range of light intensities.     

蛙离体视网膜中杆系统对锥系统的抑制作用

在蛙离体视网膜标本上研究了由条件刺激(F_c)引起的快速暗适应(RDA)过程中b 波的变化。当F_c超过一定强度时,以501nm 闪光为测试刺激(F_(t(501)))时b波阈值单调下降,先为锥相后为杆相,两者过渡处呈现杆-锥转折;但以654nm闪光为测试刺激时(F_(t(654))),阈值先略有下降,继而上升,之后再下降。F_(t(654))应阈值在RDA 过程中的这种升高是反映575锤系统的活动受到502杆系统抑制的结果,433杆系统对此没有明显的贡献,主要依据是:(1)F_(t(654))反应阈值的上升在时间上与暗适应曲线的杆相的出现一致;(2)在RDA 过程的锥相,反应的光谱敏感性与锥色素吸收光谱一致,而在F_(t(654))反应阈值上升到最大值时,反应的光谱敏感性与502杆色素的吸收光谱非常接近;(3)对502杆系统作用等效的F_(c(439))、F_(c(501))和F_(c(616))作用后,F_(t(654))的敏感度变化在整个RDA 过程中基本相同。     

鳜鱼视觉特性及其对捕食习性适应的研究Ⅰ．视网膜电图光谱敏感性和适应特性

采用电生理方法研究了夜行性凶猛鱼类鳜鱼视网膜电图的一般特性、光谱敏感性和适应特性。顷鱼的视网膜电图不显示典型的混合型视网膜特征。明视和暗视视网膜电图的光谱敏感曲线形状基本相同,峰值都在５３０ｎｍ处,没有出现Ｐｕｒｋｉｎｊｅ氏位移。明适应曲线仅出现下降型变化,暗适应过程异常缓慢。鳜鱼的视网膜仅存在单一的光感受系统,即暗视系统,不可能形成色觉。但级鱼视网膜具有很高的光敏感性,适于弱光视觉。     

The Giant Mottled Eel,Anguilla marmorata,Uses Blue-Shifted Rod Photoreceptors during Upstream Migration

Catadromous fishes migrate between ocean and freshwater during particular phases of their life cycle. The dramatic environmental changes shape their physiological features, e.g. visual sensitivity, olfactory ability, and salinity tolerance. Anguilla marmorata, a catadromous eel, migrates upstream on dark nights, following the lunar cycle. Such behavior may be correlated with ontogenetic changes in sensory systems. Therefore, this study was designed to identify changes in spectral sensitivity and opsin gene expression of A. marmorata during upstream migration. Microspectrophotometry analysis revealed that the tropical eel possesses a duplex retina with rod and cone photoreceptors. The λ_max of rod cells are 493, 489, and 489 nm in glass, yellow, and wild eels, while those of cone cells are 508, and 517 nm in yellow, and wild eels, respectively. Unlike European and American eels, Asian eels exhibited a blue-shifted pattern of rod photoreceptors during upstream migration. Quantitative gene expression analyses of four cloned opsin genes (Rh1f, Rh1d, Rh2, and SWS2) revealed that Rh1f expression is dominant at all three stages, while Rh1d is expressed only in older yellow eel. Furthermore, sequence comparison and protein modeling studies implied that a blue shift in Rh1d opsin may be induced by two known (N83, S292) and four putative (S124, V189, V286, I290) tuning sites adjacent to the retinal binding sites. Finally, expression of blue-shifted Rh1d opsin resulted in a spectral shift in rod photoreceptors. Our observations indicate that the giant mottled eel is color-blind, and its blue-shifted scotopic vision may influence its upstream migration behavior and habitat choice.     

Transretinal ERG Recordings from Mouse Retina: Rod and Cone Photoresponses

There are two distinct classes of image-forming photoreceptors in the vertebrate retina: rods and cones. Rods are able to detect single photons of light whereas cones operate continuously under rapidly changing bright light conditions. Absorption of light by rod- and cone-specific visual pigments in the outer segments of photoreceptors triggers a phototransduction cascade that eventually leads to closure of cyclic nucleotide-gated channels on the plasma membrane and cell hyperpolarization. This light-induced change in membrane current and potential can be registered as a photoresponse, by either classical suction electrode recording technique^1,2 or by transretinal electroretinogram recordings (ERG) from isolated retinas with pharmacologically blocked postsynaptic response components^3-5. The latter method allows drug-accessible long-lasting recordings from mouse photoreceptors and is particularly useful for obtaining stable photoresponses from the scarce and fragile mouse cones. In the case of cones, such experiments can be performed both in dark-adapted conditions and following intense illumination that bleaches essentially all visual pigment, to monitor the process of cone photosensitivity recovery during dark adaptation^6,7. In this video, we will show how to perform rod- and M/L-cone-driven transretinal recordings from dark-adapted mouse retina. Rod recordings will be carried out using retina of wild type (C57Bl/6) mice. For simplicity, cone recordings will be obtained from genetically modified rod transducin α-subunit knockout (Tα^-/-) mice which lack rod signaling⁸.     

The primary visual pathway in humans is regulated according to long-term light exposure through the action of a nonclassical photopigment

BACKGROUND: The mammalian eye shows marked adaptations to time of day. Some of these modifications are not acute responses to short-term light exposure but rely upon assessments of the photic environment made over several hours. In the past, all attempts at a mechanistic understanding have assumed that these adaptations originate with light detection by one or other of the classical photoreceptor cells (rods or cones). However, previous work has demonstrated that the mammalian eye contains non-rod, non-cone photoreceptors. This study aimed to determine whether such photoreceptors contribute to retinal adaptation. RESULTS: In the human retina, second-order processing of signals originating in cones takes significantly longer at night than during the day. Long-term light exposure at night is capable of reversing this effect. Here, we employed the cone ERG as a tool to examine the properties of the irradiance measurement pathway driving this reversal. Our findings indicate that this pathway (1) integrates irradiance measures over time periods ranging from at least 15 to 120 min; (2) responds to relatively bright light, having a dynamic range almost entirely outside the sensitivity of rods; (3) acts on the cone pathway primarily through a local retinal mechanism; and (4) detects light via an opsin:vitamin A photopigment (lambda(max) approximately 483 nm). CONCLUSIONS: A photopigment with a spectral sensitivity profile quite different from those of the classical rod and cone opsins but matching the standard profile of an opsin:vitamin A-based pigment drives adaptations of the human primary cone visual pathway according to time of day.     

Blue Cone Monochromacy: Visual Function and Efficacy Outcome Measures for Clinical Trials

Background

Blue Cone Monochromacy (BCM) is an X-linked retinopathy caused by mutations in the OPN1LW / OPN1MW gene cluster, encoding long (L)- and middle (M)-wavelength sensitive cone opsins. Recent evidence shows sufficient structural integrity of cone photoreceptors in BCM to warrant consideration of a gene therapy approach to the disease. In the present study, the vision in BCM is examined, specifically seeking clinically-feasible outcomes for a future clinical trial.

Methods

BCM patients (n = 25, ages 5–72) were studied with kinetic and static chromatic perimetry, full-field sensitivity testing, and eye movement recordings. Vision at the fovea and parafovea was probed with chromatic microperimetry.

Results

Kinetic fields with a Goldmann size V target were generally full. Short-wavelength (S-) sensitive cone function was normal or near normal in most patients. Light-adapted perimetry results on conventional background lights were abnormally reduced; 600-nm stimuli were seen by rods whereas white stimuli were seen by both rods and S-cones. Under dark-adapted conditions, 500-nm stimuli were seen by rods in both BCM and normals. Spectral sensitivity functions in the superior retina showed retained rod and S-cone functions in BCM under dark-adapted and light-adapted conditions. In the fovea, normal subjects showed L/M-cone mediation using a 650-nm stimulus under dark-adapted conditions, whereas BCM patients had reduced sensitivity driven by rod vision. Full-field red stimuli on bright blue backgrounds were seen by L/M-cones in normal subjects whereas BCM patients had abnormally reduced and rod-mediated sensitivities. Fixation location could vary from fovea to parafovea. Chromatic microperimetry demonstrated a large loss of sensitivity to red stimuli presented on a cyan adapting background at the anatomical fovea and surrounding parafovea.

Conclusions

BCM rods continue to signal vision under conditions normally associated with daylight vision. Localized and retina-wide outcome measures were examined to evaluate possible improvement of L/M-cone-based vision in a clinical trial.     

A duplex retina and the electroretinogram in the nocturnal Perodicticus potto.

The presence of cones in potto's retina has been proved beyond doubt although they are very restricted in number (1 cone for 300 rods). Morphologically, speaking there is no point in calling these cones "rudimentary" except for their slender outer segment. There are red sensitive elements in that retina at wavelengths beyond the spectral sensitivity of visual purple and it is tempting to assume that these elements are cones. The ERG evoked from these elements by red light differs from that in response to white and blue light. They dark-adapt faster than the receptors sensitive to blue and white flashes. However in some of their properties, for example fusion frequency, these cones behave like rods in other species. As these few cones seem to activate the bipolar cells nearly as effectively as the numerous rods, it is suggested that these cones may be responsible for day vision in the potto.     

Photoreceptors and visual pigments in the retina of the fully anadromous green sturgeon (<Emphasis Type="Italic">Acipenser medirostrus</Emphasis>) and the potamodromous pallid sturgeon (<Emphasis Type="Italic">Scaphirhynchus albus</Emphasis>)

Green sturgeon and pallid sturgeon photoreceptors were studied with scanning electron microscopy (SEM), microspectrophotometry and, in the case of the green sturgeon, retinal whole-mounts. The retinas of both species contain both rods and cones: cones comprise between 23% (whole-mount) and 36% (SEM) of the photoreceptors. The cone population of both species is dominated by large single cones, but a rare small single cone is also present. In both species, most rods have long outer segments of large diameter. A rod with a relatively thin outer segment is present in the pallid sturgeon retina. Mean cone packing density for the entire green sturgeon retina is 4,690±891 cones/mm², with the dorsal retina 14% more dense than the ventral. There is evidence for a horizontal visual streak just above and including the optic disc. Mean rod packing density is 16,006±1,668 rods/mm² for the entire retina, and fairly uniform throughout. Both species have rods with peak absorbance near 540 nm, as well as short-wavelength-sensitive cones (green: 464.5±0.7 nm; pallid: 439.7±3.5 nm); middle-wavelength-sensitive cones (green: 538.0±1.4 nm; pallid: 537.0±1.7 nm); and long-wavelength-sensitive cones (green: 613.9±3.0 nm; pallid: 617.8±7.6 nm).     

Persistent functional and structural retinal anomalies in newborn rats exposed to hyperoxia.

Previous studies have shown that newborn rats exposed postnatally to hyperoxia will develop a permanent impairment of the retinal function as determined with the electroretinogram (ERG). The purpose of our study was to examine whether postnatal hyperoxia equally alters the light- and dark-adapted ERGs and oscillatory potentials (OPs) as well as leads to permanent structural modification of the retina. During the first 14 days of life, cohorts of Sprague-Dawley rats were exposed to a hyperoxic environment, and ERGs were recorded at mean ages of approximately 25 and 55 days. Our results indicate that both light- and dark-adapted ERGs and OPs are already significantly altered within a few days following exposure to hyperoxia. None of the ERG and (or) OP parameters, with the exception of the a-wave, returned to normal values by 55 days of age. In fact some dark-adapted OPs were completely abolished following postnatal O2 exposure. Histological analysis revealed that the retina of rats exposed to hyperoxia failed to develop an outer plexiform layer and had a reduced count of horizontal cells, consistent with the permanent postreceptoral anomalies seen in the ERG responses. Our results suggest that postnatal hyperoxia causes a generalized retinal disorder leading to permanent structural modifications of the retinal cytoarchitecture and lasting anomalies of the rod and cone functions.     

棉铃虫蛾复眼光反应特性

用视网膜电位图(electroretinogram,ERG)技术研究了棉铃虫Helicoverpa armigera蛾暗适应过程中对单色光和白光刺激的光感受性变化。结果显示：(1)依ERG振幅大小(峰-峰值),在340～605 nm波谱内有3个大小不等的峰-主峰位于绿 黄光区562 nm,次峰在蓝光区483 nm,第3个峰在近紫外区400 nm,显示其至少有3种感受器;(2)性别、日龄及暗适应时间长短对其光谱敏感性有影响,低龄时雄蛾对单色光刺激较雌蛾敏感,高日龄时相反;1～5日龄内, 3日龄蛾的视网膜电位(ERP)值最高;随暗适应时间延长,其复眼对近紫外区400 nm敏感性明显增加;(3)一定光强度范围内,随单色光和白光光强度增强该蛾复眼的ERP值增大,初期增加较缓,中期较快,呈近似S型曲线,显示其复眼具有较强的光强度自调节和适应机制。     

Spectral sensitivity studies on the visual system of the praying mantis, Tenodera sinensis

In these studies a constant ERG response was used as a measure of visual sensitivity to different wavelengths of light. The dark-adapted compound eye of Tenodera sinensis is dominated by a single class of photoreceptors. with a major peak of sensitivity at about 510–520 nm, and with a minor peak of sensitivity in the near-ultraviolet region at about 370 nm. The dark-adapted dorsal ocellus does not contain a homogeneous population of sensory receptors. The sensitivity function of the dark-adapted ocellus to longer wavelength light (yellow and red) is determined by a single receptor with a major peak of sensitivity in the green at 510–520 nm with some sensitivity in the near-ultraviolet. Sensitivity at shorter wavelengths (near-ultraviolet and blue), however, involves the stimulation of both this and a near-ultraviolet-sensitive receptor with a maximum sensitivity at about 370 nm. Anatomically, the sensory cells of the dorsal ocellus of Tenodera were determined histologically to be grouped into two distinct regions, each group making its own separate contribution to the ocellar nerve. This may represent the separation of two different photoreceptor types in the ocellus of the mantis.     

Rod sensitivity of neonatal mouse and rat

We have measured the sensitivity of rod photoreceptors isolated from overnight dark-adapted mice of age P12 (neonate) through P45 (adult) with suction-pipette recording. During this age period, the dark current increased roughly in direct proportion to the length of the rod outer segment. In the same period, the flash sensitivity of rods (reciprocal of the half-saturating flash intensity) increased by approximately 1.5-fold. This slight developmental change in sensitivity was not accentuated by dark adapting the animal for just 1 h or by increasing the ambient luminance by sixfold during the prior light exposure. The same small, age-dependent change in rod sensitivity was found with rat. After preincubation of the isolated retina with 9-cis-retinal, neonatal mouse rods showed the same sensitivity as adult rods, suggesting the presence of a small amount of free opsin being responsible for their lower sensitivity. The sensitivity of neonate rods could also be increased to the adult level by dark adapting the animal continuously for several days. By comparing the sensitivity of neonate rods in darkness to that of adult rods after light bleaches, we estimated that approximately 1% of rod opsin in neonatal mouse was devoid of chromophore even after overnight dark adaptation. Overall, we were unable to confirm a previous report that a 50-fold difference in rod sensitivity existed between neonatal and adult rats.     

SPECTRAL SENSITIVITY IN TWO SPECIES OF PINNIPEDS (PHOCA VITULINA AND OTARIA FLAVESCENS)

Spectral sensitivity was measured in air in the light adapted state in two harbor seals and a South American sea lion using a behavioral training technique. Increment thresholds were determined in a spectral range from 390 nm to 670 nm in a simultaneous two‐choice discrimination task. The spectral sensitivity curves show two maxima in sensitivity, one main peak with a maximum around 500 nm in the harbor seal and around 550 nm in the South American sea lion, and a second, smaller peak with a maximum in the range of 410 nm in both species. The broad shape and the position of the maximum of the spectral sensitivity curve of the harbor seals suggests that even under photopic conditions both rods and cones are contributing to the measurements since harbor seals possess only one cone type. The maximum sensitivity in the green part of the spectrum may indicate an adaptation to a specific underwater environment.     

Occupancy of the chromophore binding site of opsin activates visual transduction in rod photoreceptors

The retinal analogue beta-ionone was used to investigate possible physiological effects of the noncovalent interaction between rod opsin and its chromophore 11-cis retinal. Isolated salamander rod photoreceptors were exposed to bright light that bleached a significant fraction of their pigment, were allowed to recover to a steady state, and then were exposed to beta-ionone. Our experiments show that in bleach-adapted rods beta-ionone causes a decrease in light sensitivity and dark current and an acceleration of the dim flash photoresponse and the rate constants of guanylyl cyclase and cGMP phosphodiesterase. Together, these observations indicate that in bleach-adapted rods beta-ionone activates phototransduction in the dark. Control experiments showed no effect of beta-ionone in either fully dark-adapted or background light-adapted cells, indicating direct interaction of beta-ionone with the free opsin produced by bleaching. We speculate that beta-ionone binds specifically in the chromophore pocket of opsin to produce a complex that is more catalytically potent than free opsin alone. We hypothesize that a similar reaction may occur in the intact retina during pigment regeneration. We propose a model of rod pigment regeneration in which binding of 11-cis retinal to opsin leads to activation of the complex accompanied by a decrease in light sensitivity. The subsequent covalent attachment of retinal to opsin completely inactivates opsin and leads to the recovery of sensitivity. Our findings resolve the conflict between biochemical and physiological data concerning the effect of the occupancy of the chromophore binding site on the catalytic potency of opsin. We show that binding of beta-ionone to rod opsin produces effects opposite to its previously described effects on cone opsin. We propose that this distinction is due to a fundamental difference in the interaction of rod and cone opsins with retinal, which may have implications for the different physiology of the two types of photoreceptors.     

Photoreceptor types and their relation to the spectral and polarization sensitivities of clupeid fishes

We used compound action potential recordings from the optic nerve of anesthetized live fish to study the spectral and polarization sensitivities of the northern anchovy and the Pacific herring. The photoreceptor structure and cone mosaic type of the (illuminated) central retina was studied by microscopy. Both species showed a single peak spectral photopic sensitivity function with λ_max= 500 nm for the northern anchovy and λ_max= 520 nm for the herring. However, only the northern anchovy exhibited polarization sensitivity; the response was 180° periodic with maximum sensitivity to horizontal polarization. Similar to the bay anchovy (Fineran and Nicol 1978), the central retina of the northern anchovy showed bifid cone units with cone lamellae parallel to the cones' lengths. The herring, on the other hand, had twin cones arranged in rows with the same orientation and tangentially arranged lamellae. Our results support the hypothesis that bifid cone units act as orthogonal dichroic filters rendering anchovies polarization sensitive. The lack of polarization sensitivity in the herring suggests that twin cones may not be used in polarization sensitivity or that one orientation of polarization receptors is insufficient for the animal to detect polarization direction. Accepted: 8 December 1997