Red Oil Droplets Research Articles

Cone distribution and visual resolution of the yellow-legged gull, Larus michahellis (Naumann, 1840).

The morphological characteristics of the yellow-legged gull's photoreceptors and cone distribution were studied using light and electron microscopy. In wholemount fresh retinas, five different coloured oil droplets located in the cone inner segments could be seen and characterized by colour, diameter and stratification. The photoreceptors were classified by comparing the fresh and fixed vertical sections under a light and electron microscope. Rods were easily distinguished from cones based on the outer segment morphology and the absence of oil droplets in their inner segments. Four types of single cones were associated with red, yellow, colourless and transparent oil droplets. Unequal double cones comprised a long principal member with a green oil droplet and an accessory short member containing a green microdroplet which was highly electron-dense under electron microscopy. The different types of oil droplets were counted from microphotographs of fresh retinal samples in 20 regions. The density, percentage and diameter of the oil droplets were determined. The results showed that central regions had the highest oil droplet density which decreased towards the retinal periphery in all quadrants. Moreover, the oil droplet density was higher in the dorsotemporal quadrant than in other retinal regions. The average density of the red oil droplets was highest in the central areas, whereas colourless oil droplets had the highest density throughout the retina. In contrast, transparent oil droplets had the lowest density across all the regions of the retina. Finally, the retinal resolution was 52.61 cycles/degree. It was calculated using the posterior nodal distance and the oil droplet diameter. The work concludes by discussing the significance of the relative proportion of different cone types across the retina.

Owls lack UV-sensitive cone opsin and red oil droplets, but see UV light at night: Retinal transcriptomes and ocular media transmittance

Most diurnal birds have cone-dominated retinae and tetrachromatic colour vision based on ultra-violet/violet-sensitive UV/V cones expressing short wavelength-sensitive opsin 1 (SWS1), S cones expressing short wavelength-sensitive opsin 2 (SWS2), M cones expressing medium wavelength-sensitive opsin (RH2) and L cones expressing long wavelength-sensitive opsin (LWS). Double cones (D) express LWS but do not contribute to colour vision. Each cone is equipped with an oil droplet, transparent in UV/V cones, but pigmented by carotenoids: galloxanthin in S, zeaxanthin in M, astaxanthin in L and a mixture in D cones. Owls (Strigiformes) are crepuscular or nocturnal birds with rod-dominated retinae and optical adaptations for high sensitivity. For eight species, the absence of functional SWS1 opsin has recently been documented, functional RH2 opsin was absent in three of these. Here we confirm the absence of SWS1 transcripts for the Long-eared owl (Asio otus) and demonstrate its absence for the Short-eared owl (Asio flammeus), Tawny owl (Strix aluco) and Boreal owl (Aegolius funereus). All four species had transcripts of RH2, albeit with low expression. All four species express all enzymes needed to produce galloxanthin, but lack CYP2J19 expression required to produce astaxanthin from dietary precursors. We also present ocular media transmittance of the Eurasian eagle owl (Bubo bubo) and Short-eared owl and predict spectral sensitivities of all photoreceptors of the Tawny owl. We conclude that owls, despite lacking UV/V cones, can detect UV light. This increases the sensitivity of their rod vision allowing them, for instance, to see UV-reflecting feathers as brighter signals at night.

M to L cone ratios determine eye sizes and baseline refractions in chickens

Following a hypothesis raised by M. and J. Neitz, Seattle, we have tested whether the abundance and the ratio of Long wavelength-sensitive (L) to Middle wavelength-sensitive (M) cones may affect eye size and development of myopia in the chicken. Fourteen chickens were treated with frosted plastic diffusers in front of one eye on day 10 post-hatching for a period of 7 days to induce deprivation myopia. Ocular dimensions were measured by A-scan ultrasonography at the beginning and at the end of the treatment and development of refractive state was tracked using infrared photorefraction. At the end of the treatment period, L and M cone densities and ratios were analyzed in retinal flat mounts of both myopic and control eyes, using the red and yellow oil droplets as markers. Because large numbers of cones were counted (>10000), software was written in Visual C++ for automated cone detection and density analysis. (1) On average, 9.7 ± 1.7D of deprivation myopia was induced in 7 days (range from 6.8D to 13.7D) with an average increase in axial length by 0.65 ± 0.20 mm (range 0.42 mm–1.00 mm), (2) the increase in vitreous chamber depth was correlated with the increase in myopic refractive error, (3) average central M cone densities were 10,498 cells/mm2, and L cone densities 9574 cells/mm2. In the periphery, M cone densities were 6343 cells/mm2 and L cones 5735 cells/mm2 (4) M to L cone ratios were highly correlated in both eyes of each animal (p < 0.01 in all cases), (5) the most striking finding was that ratios of M to L cones were significantly correlated with vitreous chamber depths and refractive states in the control eyes with normal vision, both in the central and peripheral retinas (p < 0.05 to p < 0.01), (6) M to L cone ratios did however not predict the amount of deprivation myopia that could be induced. M and L cone ratios are most likely genetically determined in each animal. The more L cones, the deeper the vitreous chambers and the more myopic were the refractions in eyes. M to L cone ratios may determine the set point of emmetropization and thereby ultimately the probability of becoming myopic. Deprivation myopia was not determined by M to L cone ratios.

Evolution of CYP2J19, a gene involved in colour vision and red coloration in birds: positive selection in the face of conservation and pleiotropy

BackgroundExaggerated signals, such as brilliant colours, are usually assumed to evolve through antagonistic coevolution between senders and receivers, but the underlying genetic mechanisms are rarely known. Here we explore a recently identified “redness gene”, CYP2J19, that is highly interesting in this context since it encodes a carotenoid-modifying enzyme (a C4 ketolase involved in both colour signalling and colour discrimination in the red (long wavelength) spectral region.)ResultsA single full-length CYP2J19 was retrieved from 43 species out of 70 avian genomes examined, representing all major avian clades. In addition, CYP2J19 sequences from 13 species of weaverbirds (Ploceidae), seven of which have red C4-ketocarotenoid coloration were analysed. Despite the conserved retinal function and pleiotropy of CYP2J19, analyses indicate that the gene has been positively selected throughout the radiation of birds, including sites within functional domains described in related CYP (cytochrome P450) loci. Analyses of eight further CYP loci across 25 species show that positive selection is common in this gene family in birds. There was no evidence for a change in selection pressure on CYP2J19 following co-option for red coloration in the weaverbirds.ConclusionsThe results presented here are consistent with an ancestral conserved function of CYP2J19 in the pigmentation of red retinal oil droplets used for colour vision, and its subsequent co-option for red integumentary coloration. The cause of positive selection on CYP2J19 is unclear, but may be partly related to compensatory mutations related to selection at the adjacent gene CYP2J40.

Expression of a carotenoid-modifying gene and evolution of red coloration in weaverbirds (Ploceidae).

Red carotenoid colours in birds are widely assumed to be sexually selected quality indicators, but this rests on a very incomplete understanding of genetic mechanisms and honesty-mediating costs. Recent progress was made by the implication of the gene CYP2J19 as an avian carotenoid ketolase, catalysing the synthesis of red C4-ketocarotenoids from yellow dietary precursors, and potentially a major mechanism behind red coloration in birds. Here, we investigate the role of CYP2J19 in the spectacular colour diversification of African weaverbirds (Ploceidae), represented by five genera and 16 species: eight red, seven yellow and one without carotenoid coloration. All species had a single copy of CYP2J19, unlike the duplication found in the zebra finch, with high expression in the retina, confirming its function in colouring red oil droplets. Expression was weak or undetected in skin and follicles of pigment-depositing feather buds, as well as in beaks and tarsi, including those of the red-billed quelea. In contrast, the hepatic (liver) expression of CYP2J19 was consistently higher (>14-fold) in seven species with C4-ketocarotenoid coloration than in species without (including one red species), an association strongly supported by a phylogenetic comparative analysis. The results suggest a critical role of the candidate ketolase, CYP2J19, in the evolution of red C4-ketocarotenoid colour variation in ploceids. As ancestral state reconstruction suggests that ketocarotenoid coloration has evolved twice in this group (once in Euplectes and once in the Quelea/Foudia clade), we argue that while CYP2J19 has retained its ancestral role in the retina, it has likely been co-opted for red coloration independently in the two lineages, via increased hepatic expression.

Independent pseudogenization of CYP2J19 in penguins, owls and kiwis implicates gene in red carotenoid synthesis

Carotenoids have important roles in bird behavior, including pigmentation for sexual signaling and improving color vision via retinal oil droplets. Yellow carotenoids are diet-derived, but red carotenoids (ketocarotenoids) are typically synthesized from yellow precursors via a carotenoid ketolase. Recent research on passerines has provided evidence that a cytochrome p450 enzyme, CYP2J19, is responsible for this reaction, though it is unclear if this function is phylogenetically restricted. Here I provide evidence that CYP2J19 is the carotenoid ketolase common to Aves using the genomes of 65 birds and the retinal transcriptomes of 15 avian taxa. CYP2J19 is functionally intact and robustly transcribed in all taxa except for several species adapted to foraging in dim light conditions. Two penguins, an owl and a kiwi show evidence of genetic lesions and relaxed selection in their genomic copy of CYP2J19, and six owls show evidence of marked reduction in CYP2J19 retinal transcription compared to nine diurnal avian taxa. Furthermore, one of the owls appears to transcribe a CYP2J19 pseudogene. Notably, none of these taxa are known to use red carotenoids for sexual signaling and several species of owls and penguins represent the only birds known to completely lack red retinal oil droplets. The remaining avian taxa belong to groups known to possess red oil droplets, are known or expected to deposit red carotenoids in skin and/or plumage, and/or frequently forage in bright light. The loss and reduced expression of CYP2J19 is likely an adaptation to maximize retinal sensitivity, given that oil droplets reduce the amount of light available to the retina.

Seeing red to being red: conserved genetic mechanism for red cone oil droplets and co-option for red coloration in birds and turtles.

Avian ketocarotenoid pigments occur in both the red retinal oil droplets that contribute to colour vision and bright red coloration used in signalling. Turtles are the only other tetrapods with red retinal oil droplets, and some also display red carotenoid-based coloration. Recently, the CYP2J19 gene was strongly implicated in ketocarotenoid synthesis in birds. Here, we investigate CYP2J19 evolution in relation to colour vision and red coloration in reptiles using genomic and expression data. We show that turtles, but not crocodiles or lepidosaurs, possess a CYP2J19 orthologue, which arose via gene duplication before turtles and archosaurs split, and which is strongly and specifically expressed in the ketocarotenoid-containing retina and red integument. We infer that CYP2J19 initially functioned in colour vision in archelosaurs and conclude that red ketocarotenoid-based coloration evolved independently in birds and turtles via gene regulatory changes of CYP2J19 Our results suggest that red oil droplets contributed to colour vision in dinosaurs and pterosaurs.

Morphology, Characterization and Distribution of Retinal Photoreceptors in the South American (Lepidosiren paradoxa) and Spotted African (Protopterus dolloi) Lungfishes

Lungfishes are the closest living relatives of the ancestors to all terrestrial vertebrates and have remained relatively unchanged since the early Lochkovin period (410 mya). Lungfishes, therefore, represent a critical stage in vertebrate evolution and their sensory neurobiology is of considerable interest. This study examines the ultrastructure of the retina of two species of lungfishes: the South American lungfish, Lepidosiren paradoxa and the spotted African lungfish, Protopterus dolloi in an attempt to assess variations in photoreception in these two ancient groups of sarcopterygian (lobe-finned) fishes. In juvenile P. dolloi, the retina contains one rod and two cone photoreceptor types (one containing a red oil droplet), while only one rod and one cone photoreceptor type is present in adult L. paradoxa. Both species lack double cones. The large size and inclusion of oil droplets in both species apart from one of the cone photoreceptor types in P. dolloi suggests that L. paradoxa and P. dolloi are adapted for increasing sensitivity. However, the complement of photoreceptor types suggests that there may be a major difference in the capacity to discriminate colour (dichromatic and monochromatic photoreception in P. dolloi and L. paradoxa, respectively). This study suggests that the visual needs of these two species may differ.

Development of a simple experimental method for the determination of the liquid field velocity in conical and cylindrical hydrocyclones

The objective of this work was to propose a new and simple method to understand the hydrodynamic phenomena that occur within two types of hydrocyclones, cylindrical and conical. Using a PCO 1200 hs digital high-speed camera, this method involved the following of a little red oil droplet that had been injected by a small syringe into the hydrocyclone at different zones. The trajectory line of the injected droplets and velocity of the water were investigated, in terms of the tangential and axial velocities of the water and the average velocity gradient in the hydrocyclone. In both hydrocyclones, if the oil droplets were injected at the center of the hydrocyclone, they went down to the bottom part of the hydrocyclone. In contrast, if the droplets were injected close to the wall of the hydrocyclone, they rose rapidly to the upper part of the hydrocyclone. Moreover, the tangential velocity of water in both hydrocyclones presented average velocity gradient of the water at the center of the hydrocyclone which was significantly lower than the average velocity gradient of the water close to the wall of hydrocyclone. Therefore, it could be concluded that the external vortex close to the wall of the hydrocyclone was stronger than the internal vortex at the center of the hydrocyclone. These obtained results are in agreement with those observations in the literatures. In conclusion, we propose this simple experimental method as an effective means of understanding the hydrodynamic phenomena that occurs within hydrocyclones and so improving the hydrocyclone design for further applications.

Visual ecology of the Australian lungfish (Neoceratodus forsteri)

BackgroundThe transition from water to land was a key event in the evolution of vertebrates that occurred over a period of 15–20 million years towards the end of the Devonian. Tetrapods, including all land-living vertebrates, are thought to have evolved from lobe-finned (sarcopterygian) fish that developed adaptations for an amphibious existence. However, while many of the biomechanical and physiological modifications necessary to achieve this feat have been studied in detail, little is known about the sensory adaptations accompanying this transition. In this study, we investigated the visual system and visual ecology of the Australian lungfish Neoceratodus forsteri, which is the most primitive of all the lungfish and possibly the closest living relative to the ancestors of tetrapods.ResultsJuvenile Neoceratodus have five spectrally distinct retinal visual pigments. A single type of rod photoreceptor contains a visual pigment with a wavelength of maximum absorbance (λmax) at 540 nm. Four spectrally distinct single cone photoreceptors contain visual pigments with λmax at 366 (UVS), 479 (SWS), 558 (MWS) and 623 nm (LWS). No double cones were found. Adult lungfish do not possess UVS cones and, unlike juveniles, have ocular media that prevent ultraviolet light from reaching the retina. Yellow ellipsoidal/paraboloidal pigments in the MWS cones and red oil droplets in the LWS cones narrow the spectral sensitivity functions of these photoreceptors and shift their peak sensitivity to 584 nm and 656 nm, respectively. Modelling of the effects of these intracellular spectral filters on the photoreceptor colour space of Neoceratodus suggests that they enhance their ability to discriminate objects, such as plants and other lungfishes, on the basis of colour.ConclusionThe presence of a complex colour vision system based on multiple cone types and intracellular spectral filters in lungfishes suggests that many of the ocular characteristics seen in terrestrial or secondarily aquatic vertebrates, such as birds and turtles, may have evolved in shallow water prior to the transition onto land. Moreover, the benefits of spectral filters for colour discrimination apply equally to purely aquatic species as well as semi-aquatic and terrestrial animals. The visual system of the Australian lungfish resembles that of terrestrial vertebrates far more closely than that of other sarcopterygian fish. This supports the idea that lungfishes, and not the coelacanth, are the closest living relatives of the ancestors of tetrapods.

Topography of retinal photoreceptor cells in the Jungle Crow (Corvus macrorhynchos) with emphasis on the distribution of oil droplets

Previously, we analyzed the regional specialization of ganglion cells in the retina of the Jungle Crow (Corvus macrorhynchos). The present study extends this analysis by determining the numbers and regional distribution of photoreceptor cells and the different colored oil droplets. The numbers of the different colored oil droplets were counted using color microphotographs of fresh retinas. Estimation of the numbers of photoreceptor cells was made using Nissl-stained retinal whole mounts. The estimated total number of photoreceptor cells in the whole mount retina was approximately 17,933,788 (N=4). The peak density of photoreceptor cells was 92,109/mm2 in the central area of the retina, followed by temporal and nasal areas, respectively. Four types of oil droplet were identified on the basis of their color: red, orange, green and translucent. The central retina had a significantly higher population of oil droplets, with a density of 91,202/mm2; this sharply declined at the peripheral retina. The lowest observed density was 13,192/mm2 in the dorsal retina. The densities and sizes of the different colored oil droplets were inversely correlated across the retina. The average sizes of red, orange, green and translucent oil droplets had 6 μm2, 4 μm2, 7 μm2 and 4 μm2, respectively. At the periphery of the retina, the oil droplets were significantly larger than in the central area. The sizes of the different colored oil droplets also varied within the same area. Orange colored oil droplets (33%) were the most common type in the central area, whereas green colored oil droplets were the most frequent in other areas. The results of the present study show that photoreceptor cells and oil droplets are differentially distributed in the retina. From these results, we conclude that the area of high cell density, matched by high neuron densities of the ganglion cell layer, corresponds to the site of acute color vision in the crow retina.

An Anatomical Study of the Visual Capabilities of the Green Turtle, Chelonia mydas

Several aspects of vision in juvenile and adult Green Turtles (Chelonia mydas) are examined, with special reference to retinal anatomy such as oil droplet topography, transmission electron microscopy of photoreceptors, spectral transmission measurements of the ocular media (cornea, lens, and vitreous humor), and measurements of focal length and optical sensitivity. A detailed study of the distribution of the different color classes of oil droplets shows that all oil droplets are found in high concentrations (>1000 mm−2) in the central/temporal parts of the retina. Red oil droplets were the largest, followed by yellow and clear. Oil droplet size varied in different parts of the retina. On average, red oil droplets were found in fewer numbers compared to yellow and clear oil droplets. Two types of clear oil droplets were identified: those that fluoresced under UV illumination and those that did not. We found that the majority (78.5%) of colorless oil droplets fluoresced when viewed under UV light. Spectral transmission measurements of the ocular media show that wavelengths to approximately 325 nm are transmitted. This may suggest ultraviolet (UV) vision in Green Turtles. The optical sensitivity of the Green Turtle eye was relatively low, suggesting an adaptation to high light intensities commonly experienced by this species.

Morphology, characterization, and distribution of retinal photoreceptors in the Australian lungfishNeoceratodus forsteri (Krefft, 1870)

The Australian lungfish Neoceratodus forsteri (Dipnoi) is an ancient fish that has a unique phylogenetic relationship among the basal Sarcopterygii. Here we examine the ultrastructure, histochemistry, and distribution of the retinal photoreceptors using a combination of light and electron microscopy in order to determine the characteristics of the photoreceptor layer in this living fossil. Similar proportions of rods (53%) and cones (47%) reveal that N. forsteri optimizes both scotopic and photopic sensitivity according to its visual demands. Scotopic sensitivity is optimized by a tapetum lucidum and extremely large rods (18.62 +/- 2.68 microm ellipsoid diameter). Photopic sensitivity is optimized with a theoretical spatial resolving power of 3.28 +/- 0.66 cycles degree(-1), which is based on the spacing of at least three different cone types: a red cone containing a red oil droplet, a yellow cone containing a yellow ellipsoidal pigment, and a colorless cone containing multiple clear oil droplets. Topographic analysis reveals a heterogeneous distribution of all photoreceptor types, with peak cone densities predominantly found in temporal retina (6,020 rods mm(-2), 4,670 red cones mm(-2), 900 yellow cones mm(-2), and 320 colorless cones mm(-2)), but ontogenetic changes in distribution are revealed. Spatial resolving power and the diameter of all photoreceptor types (except yellow cones) increases linearly with growth. The presence of at least three morphological types of cones provides the potential for color vision, which could play a role in the clearer waters of its freshwater environment.

Novel identification of the different types of cones in the retina of the chicken.

1. Although single cones and double cones in the chicken retina had been documented for more than 30 years, the exact morphology of these cells had never been studied by the scanning electronmicroscopy. In this brief report, we present the evidence for the first time the existence of two types of single cones and three types of double cones (termed as types A, B. and C), each with distinct morphology. 2. The proportion of type A:type B:type C double cones, as estimated from the midperipheral and central regions of our scanning specimens, was about 30%:50%:20%. 3. Based on the literature data that red oil droplets reside in single cones and yellow oil droplets in chief cones of the double cones, the proportion of single to double cones was deciphered and the relative proportion was estimated to be 1:0.91 in the central region, 1:0.92 in the midperiphery, and 1:0.84 in the periphery.

Photoreceptors and visual pigments in the red-eared turtle, Trachemys scripta elegans.

Absorbance spectra of cone outer segments and oil droplets were recorded microspectrophotometrically in the retina of the red-eared turtle, Trachemys scripta elegans. There are four cone visual pigments, with lambda(max) = 617 nm (red sensitive), 515 nm (green sensitive), 458 nm (blue sensitive), and 372 nm (UV-sensitive). The red-sensitive pigment resides in single cones with red or orange oil droplets, and in both members of double cones. The principal member of the double cone contains an orange oil droplet, and the accessory member is droplet free. The green-sensitive pigment is situated in single cones with orange/dark yellow droplets. The blue-sensitive pigment is combined with the UV-absorbing oil droplet in single cones. The UV-sensitive pigment resides in single cones with clear oil droplets that exhibited virtually no absorbance down to 325 nm. Thus, seven types of cones can be identified based on their morphology, oil droplet color, and the visual pigment absorbance. At the moment, this is the most complex cone system described for vertebrates.

Comparative Histophysiological Study of Oil Droplets in the Avian Retina

The colored oil droplets in the retina of 30 avian species with different ecological habits and different taxonomic positions were examined by microspectrophotometry and histology, and their role or significance was conjectured. In diurnal birds such as the Night Heron, Jungle Crow, and Japanese Lesser Sparrow Hawk, four kinds of colored oil droplets, red, yellow, orange, and pale green, were discriminated ; whereas in nocturnal birds, only pale green oil droplets were found. Cluster analysis of the distribution of droplets showed that the color of the oil droplets in the 13 species of wild birds correlates with their ecological behavior. The absorption spectra of the red, yellow, and orange oil droplets displayed only minor interspecific differences. The spectra of the pale green droplets differed significantly between the Eastern House Swallow and the Japanese Jungle Nightjar. In the King Penguin, the retina contained only pale green droplets, whose spectrum was completely different from that of other diurnal birds. The upper half of the retina of the Japanese Jungle Nightjar was lined characteristically with the tapetum lucidum of nocturnal birds ; and the under half, with the diurnal birds' pigmented epithelium. Our findings suggest that each colored oil droplet has specific advantage for visual perception and that each bird has the retina that best matches its natural environment and feeding behavior in each habitat.

Colour Discrimination of Two Passerine Bird Species in the Munsell System

Two male Yellow Wagtails Motacilla f. flava and one male Bluethroat Luscinia s. svecica were successively trained to ten colour papers, each representing one of the ten colour families in the Munsell 40 hue system. Discrimination was tested between the training hue and four neighbouring hues under standardized illumination. Yellow Wagtail hue discrimination was best in the red, orange and yellow families and less good towards shorter wavelengths and for red-purple hues. The Bluethroat discriminated best in the orange, purple-blue and purple families. Both species could memorize and make statistically significant discriminations between c. 20 hues. Their best discrimination occurred in spectral regions corresponding to their own sexually dimorphic plumage. Brightness (value) and saturation (chroma) discrimination was studied by training the same birds to the middle value/chroma of seven Munsell hues. The birds' value discrimination was poor: they learned to reject the darkest value(s) but not the brighter ones. Both species could discriminate between the less saturated chromas but they chose more saturated colours almost as frequently as the correct ones. Between the species there were differences particularly in chroma discrimination similar to those in hue discrimination. The Bluethroat's better hue discrimination in the short wavelength end of the spectrum (violet and blue) may tentatively be connected with the fact that this species has a larger proportion of cones sensitive to shorter wavelengths (about 75% of cones with colourless or pale greenish droplets), while in the Yellow Wagtail about 50% of the cones have red or yellow-orange oil droplets transmitting light efficiently only in the long wavelength part of the spectrum.

Immunohistochemical localization of iodopsin in the retina of the chicken and Japanese quail

Localization of iodopsin in the retina of the chicken and Japanese quail was investigated immunohistochemically with the use of monoclonal antibodies (R1-R4) highly specific for R-photopsin (protein moiety of iodopsin). In paraffin sections of the retina, the outer segments of double cones (principal and accessory cones) and of one particular type of single cones were labeled with the antibodies. In addition, reticular cytoplasmic structures, probably representing the Golgi apparatus in a position close to the vitreous pole of the paraboloid and to the outer limiting membrane were intensely stained in the cone cells bearing an immunoreactive outer segment. In whole-mount preparations, 5 types of cone cells were identified according to the color of oil droplets, i.e., red, yellow, pale-green (principal member of double cones), pale-blue and clear, in addition to a sixth type devoid of an oil droplet (accessory member of double cones). The immunohistochemical analysis of the preparations revealed that R-photopsin (suggesting the presence of iodopsin) is localized in the outer segments of both the principal and accessory members of double cones, and the population of single cones displaying a red oil droplet. Other cones endowed with a yellow, blue or clear oil droplet were not labeled with the antibodies used. Similar results were obtained in the retina of the Japanese quail.

Monoclonal antibody labels both rod and cone outer segments of turtle photoreceptors

We have studied the immunoreactivity of turtle photoreceptors to a monoclonal antibody (MAb 15–18) which binds to the external loop connecting bovine rhodopsin helices IV–V. Three chromatic types of cone photoreceptors were identified by the presence and color of oil droplets. MAb 15–18 intensely labeled the outer segments of both rods and green cones. In addition, a weak cross-reactivity was also found in the outer segments of red cones having a pale-green oil droplet, and of blue cones. Other morphological subtypes of red cones. cones with a red oil droplet and both members of double cones, showed no labeling. Our results indicate that rhodopsin and green cone opsin have a similar antigenic determinant, and that two different structural forms of red cone opsin may be present in the turtle retina.

Relation of spectral types to oil droplets in cones of turtle retina.

The spectral sensitivities and color of oil droplets of cone photoreceptors in the retina of the red-eared turtle (Pseudemys scripta elegans) were investigated by intracellular recording and injections of Lucifer yellow dye. Six morphological types of cones could be distinguished by the color of the oil droplets located in the outermost inner segments. Single cones containing either red or pale green oil droplets were sensitive to red light, cones with yellow oil droplets to green, and cones with clear oil droplets to blue. Contrary to previous reports, both principal and accessory members of double cones were sensitive to red, and no diffusion of dye was detected between the two apposed members. Thus, the oil droplets provide a reliable morphological basis for further investigation of the neuroanatomical networks underlying the processing of color information in the vertebrate retina.