Pigmentation formation and expression analysis of tyrosinase in Siniperca chuatsi.

Animal pigmentation primarily depends on the presence and mixing ratio of chromatophores, functioning in animal survival and communication. For the benthic and carnivorous Siniperca chuatsi, pigmentation pattern is key to concealment and predation. In this study, the formation, distribution, and main pattern of chromatophores were observed in the embryos, larvae, skins, and visceral tissues from S. chuatsi. Melanophores were firstly visualized in the yolk sac at segmentation stage, and then they were migrated to the whole body and further clustered into the black stripes, bands, and patches. In adult S. chuatsi, the head, black band, and body side skins mainly contained melanophores, showing as deep or light black. The abdomen skin mainly contained iridophores, showing as silvery. In the eye, the pigment layers were located in the epithelial layers of iris and retina and shown as black. Then, the pigmentation-related gene, tyrosinase gene from S. chuatsi (Sc-tyr) was analyzed by bioinformatics and quantitative methods. The Sc-tyr gene encoded a protein with 540 amino acids (Sc-TYR). The Sc-TYR contained two copper ion binding sites, which were coordinated by six conserved histidines (H182, H205, H214, H366, H370, H393) and necessary for catalytic activity. The Sc-TYR was well conserved compared with TYR of various species with higher degree of sequence similarity with other fishes (77.6-98.3%). The qRT-PCR test showed that the Sc-tyr mRNA reached the peak value at segmentation stage in the embryo development, the black skins displayed a higher expression level than that in silvery skin, and the eye had the highest expression level compared with other tissues. Further research on enzyme activity showed that the expression patterns of tyrosinase activity were similar to that of the Sc-tyr mRNA. Comparing with the results of molecular and phenotype, it was found that the temporal and spatial distributions of tyrosinase corresponded well with changes in pigmentation patterns and the intensity of skin melanization. This study initially explored the pigmentation formation and tyrosinase expression, which served as a foundation for further insight into the genetics mechanism of body color formation in S. chuatsi.

The eyes of lanternfishes (Myctophidae, Teleostei): novel ocular specializations for vision in dim light.

Lanternfishes are one of the most abundant groups of mesopelagic fishes in the world's oceans and play a critical role in biomass vertical turnover. Despite their importance, very little is known about their physiology or how they use their sensory systems to survive in the extreme conditions of the deep sea. In this study, we provide a comprehensive description of the general morphology of the myctophid eye, based on analysis of 53 different species, to understand better their visual capabilities. Results confirm that myctophids possess several visual adaptations for dim-light conditions, including enlarged eyes, an aphakic gap, a tapetum lucidum, and a pure rod retina with high densities of long photoreceptors. Two novel retinal specializations were also discovered. The first specialization is a fundal pigmentation in adult eyes, found within an isolated retinal region (typically central retina) composed of modified pigment epithelial cells, which we hypothesize to be the remnant of a more pronounced visual specialization important in larval stages. The second specialization is an aggregation of extracellular microtubular-like structures found within the sclerad region of the inner nuclear layer of the retina. We hypothesize that the marked interspecific differences in the hypertrophy of these microtubular-like structures may be related to inherent differences in visual function. A general interspecific variability in other parts of the eye is also revealed and examined in this study. The contribution of both ecology and phylogeny to the evolution of ocular specializations and vision in dim light are discussed.

The structure of anchovy outer retinae (Engraulididae, Clupeiformes) - a comparative light- and electron-microscopic study using museum-stored material.

The outer retinal architecture of Engraulididae is uncommon among vertebrates. In some anchovies, e.g., Anchoa, two cone types are arranged alternating in long photoreceptor chains, i.e., polycones. The cones have radially oriented outer segment lamellae in close contact with a complex guanine tapetum, most probably subserving polarization contrast vision. To clarify the distribution of the aberrant polycone architecture within the Engraulididae and to provide indications about polycone evolution, the outer retina morphology of 16 clupeoid species was investigated by light and electron microscopy, predominantly using museum-stored material. The outgroup representatives of four clupeid subfamilies (Clupeonella cultriventris, Dorosoma cepedianum, Ethmalosa fimbriata, Pellonula leonensis) show a row pattern of double cones, partially with single cones at defined positions and a pigment epithelium with lobopodial protrusions containing melanin. The pristigasterid Ilisha africana has double rows of single cones lying between linear curtains of pigment epithelium processes filled with minute crystallites and melanin concentrated near their vitreal tips. Within the Engraulididae, two main architectures are found: Coilia nasus and Thryssa setirostris have linear multiple cones or polycones separated by long pigment epithelium barriers containing tapetal crystallites and melanin in the tips (also found in Setipinna taty), whereas Anchoviella alleni, Encrasicholina heteroloba, Engraulis encrasicolus, Engraulis mordax, Lycengraulis batesii, and Stolephorus indicus exhibit the typical polycone architecture. Cetengraulis mysticetus and Lycothrissa crocodilus show cone patterns and pigment epithelium morphology differing from the other anchovy species. The sets of characters are compared, corroborated with the previous knowledge on clupeoid retinae and discussed in terms of functional morphology and visual ecology. A scenario on polycone evolution is developed that may serve as an aid for the reconstruction of engraulidid phylogeny. Furthermore, this study demonstrates the suitability of museum material for morphological studies, even at the electron microscopic level.

Redefining the limit of the outer retina in optical coherence tomography scans.

OBJECTIVE: A highly reflective layer seen in retinal optical coherence tomography (OCT) has been believed to correspond to the choriocapillaris (CHC) and retinal pigment epithelium (RPE). On gray-scale scans of OCT-2000, and on Stratus OCT, this layer by the outer retinal limit can be resolved into 2 distinct laminae. We analyzed these 2 laminae in normal and abnormal maculae to infer their anatomic correlate. DESIGN: Retrospective study. METHODS: Analysis of macular OCT scans was performed in 44 patients using OCT-2000, and in 39 patients using Stratus OCT. Thirty of these patients had no ocular disease, and their OCT was normal. The other 53 patients had several macular diseases of different etiologies. Both color and gray-scale images were analyzed. RESULTS: Macular OCT scans showed a double laminae at the level where the retina interfaces the RPE in normal subjects using both OCT-2000 and Stratus OCT. In 2-dimensional scans, this laminar structure appears as a double line. It is best distinguished on the Stratus OCT and gray-scale images of OCT-2000. This double line consisted of a thin inner line and a thicker outer line. Similar analysis in patients with macular pathology showed a discernible double line at the retina/RPE interface in at least part of the scan. However, in patients with macular hole, the area corresponding to the absent retina showed only a single line. The inner line component appeared to follow the contour of the retina. This phenomenon was also seen in eyes with neurosensory detachment secondary to central serous chorioretinopathy and other etiologies. In contrast, in macular pathologies where the outer retina did not lose contiguity with the RPE, such as in lamellar macular hole and in cystoid macular edema, the double line persisted. Software for retinal thickness measurements regularly place the outer limit of the retina at the internal aspect of the inner line, probably underestimating the retinal thickness by about 24 to 34 mum. CONCLUSIONS: A double laminar structure at the outer retina/RPE/CHC interface can be consistently distinguished on commercially available OCT of normal eyes. In eyes with macular pathology, OCT analysis of the inner lamina leads us to conclude it is most likely part of the neurosensory retina and not part of the RPE/CHC complex as previously thought.

Similarity of mRNA phenotypes of morphologically normal macular and peripheral retinal pigment epithelial cells in older human eyes.

PURPOSE: To determine the expression profiles of morphologically normal human retinal pigment epithelial (RPE) cells that originate from the macula and periphery. METHODS: Morphologically normal RPE cells from 15 human globes from donors aged 52 to 82 years old were laser capture microdissected. Total RNA from 5000 cells was SMART amplified, [33]P-labeled, and hybridized to a cDNA array containing 4325 known genes. Expression profiles were analyzed by hierarchical cluster analysis, Prediction Analysis of Microarrays (PAM), and Significance Analysis for Microarrays (SAM). Differentially expressed genes were evaluated further by real time RT-PCR. RESULTS: The overall expression profiles of RPE cells from the macula and periphery were similar. Unsupervised and supervised hierarchical cluster analysis showed that patient genotype was a stronger separating factor than topographical location. SAM analysis identified 11 genes that were underexpressed by macular RPE cells. The expression patterns of these 11 genes were confirmed by real time RT-PCR, with 5 genes reaching statistical significance. CONCLUSIONS: Whereas the overall expression profiles were similar between cells from the macula and periphery, subtle differential expression of five genes could contribute to RPE phenotypic differences based on topographic location.

Lack of type XVIII collagen results in anterior ocular defects.

Mice lacking type XVIII collagen have defects in the posterior part of the eye, including delayed regression of the hyaloid vasculature and poor outgrowth of the retinal vessels. We report here that these mice also have a fragile iris and develop atrophy of the ciliary body. The irises of Col18a1-/- mice can be seen to adhere to the lens and cornea. After the pupils begin to function, the double layer of epithelial cells separates at the apical cell contacts, leading to defoliation of its posterior pigment epithelial cell layer, and extracellular material begins to accumulate in the basement membrane zones of the iris. In contrast to the iris epithelia, where no clear signs of cellular atrophy were detected, the lack of type XVIII collagen resulted in atrophy of the pigmented epithelial cells of the ciliary body, and there were also ultrastructural abnormalities in the basement membrane zones. These changes did not lead to chronically elevated intraocular pressures, however. Our results indicate that type XVIII collagen is needed for the integrity of the epithelial basement membranes of the iris and the ciliary body and that its gene should therefore be taken into account as a new potential cause of anterior segment disorders in the eye.

Retinoprotective effect of Epithalon in campbell rats of various ages.

We studied the retinoprotective effect of Epithalon administered to the offspring of Campbell rats during postnatal ontogeny and to mothers before mating and during pregnancy. After this treatment the morphological structure and functional activity of the retina were preserved for a longer period compared to control rats (by 2 times) and to the animals receiving the peptide only during postnatal ontogeny (by 30%).

Fine structure and development of the retina of the grenadier anchovy Coilia nasus (Engraulididae, Clupeiformes).

A study of the morphogenesis of the grenadier anchovy retina was undertaken using light and electron microscopy. Five developmental stages from prelarvae 3 days after fertilization to adult fish were studied. In addition to the general morphology of the eye and retina, special emphasis was given to the development of the photoreceptors and pigment epithelium (PE). The earliest retinae showing structural features indicative of a functioning eye are pure cone retinae composed of rows of alternating long and short cones forming a transient, tesselated pattern. At this stage there is a conventional PE containing melanin. In older stages cone rows are separated by the newly formed rods and by PE wedges filled with diffusely reflecting guanine crystallites. The findings are compared with the retinae of other engraulidids and with the development of teleost retinae in general. Moreover, the observed structural changes are discussed with respect to the photic habitat conditions of these anadromous fish that move between coastal waters, estuary, and river.

High-resolution ultrasonic imaging and characterization of the ciliary body.

PURPOSE: To develop a means for noninvasive in vivo visualization of the ciliary processes using very-high-frequency (50 MHz) ultrasound and to develop quantitative morphologic descriptors that may relate to physiologic function. METHODS: The region of the ciliary body was scanned with very-high-frequency ultrasound, both in rabbits and in normal human subjects. Data were acquired in a series of planes so that the spacing between them was less than the beam width of the transducer in its focal plane. Three-dimensional perspective images were constructed, representing the anatomy of the angle region, including the ciliary processes. The automatically detected boundaries of the ciliary processes were analyzed to compute their periphery, area, shape factor, and fractal dimension. These measures were compared between the human and the rabbit eye and analyzed for periodicities related to the spacing of successive processes. RESULTS: Three-dimensional images allowed visualization of the radial arrangement of the processes. All biometric descriptors were significantly different between the rabbit and human eye and showed periodicities consistent with spacing between processes. CONCLUSIONS: The methods described in this report are sensitive descriptors of the state of the ciliary processes. These techniques may be of value in measurement of changes in the ciliary body associated with disease, medical therapy, and aging.

Macular pigment optical density measurement: a novel compact instrument.

A compact device to derive the optical density of human macular pigment (MP) using heterochromatic flicker photometry is described. The validity of the system is assessed by measuring the optical density spectra of MP in 12 healthy subjects and comparing this with well-established previously published values. The mean spectral absorbance characteristics of MP across subjects corresponds well with accepted values. As reported in other studies, our measurements show a wide variation of MP optical densities between individuals. In our technique within-subject variability is low; standard deviations are between 0.025 and 0.15 in most cases. The overall optical density of MP ranged from 0.08 to 0.84 with a mean of 0.496 and standard deviation of 0.257 at 460 nm. The stimulus size was 0.95 degrees. The unique feature of the technique is that it allows free viewing (not Maxwellian View) of the stimulus, it can be conducted easily and quickly and does not need frequent re-calibration.

Relation among serum and tissue concentrations of lutein and zeaxanthin and macular pigment density.

BACKGROUND: Lutein and zeaxanthin are the only carotenoids in the macular region of the retina (referred to as macular pigment [MP]). Foods that are rich in lutein and zeaxanthin can increase MP density. Response to dietary lutein and zeaxanthin in other tissues has not been studied. OBJECTIVE: The objective of this study was to examine tissue responses to dietary lutein and zeaxanthin and relations among tissues in lutein and zeaxanthin concentrations. DESIGN: Seven subjects consumed spinach and corn, which contain lutein and zeaxanthin, with their daily diets for 15 wk. At 0, 4, 8, and 15 wk and 2 mo after the study, serum, buccal mucosa cells, and adipose tissue were analyzed for carotenoids, and MP density was measured. RESULTS: Serum and buccal cell concentrations of lutein increased significantly from baseline during dietary modification. Serum zeaxanthin concentrations were greater than at baseline only at 4 wk, whereas buccal cell and adipose tissue concentrations of zeaxanthin did not change. Adipose tissue lutein concentrations peaked at 8 wk. Changes in adipose tissue lutein concentration were inversely related to the changes in MP density, suggesting an interaction between adipose tissue and retina in lutein metabolism. To investigate the possibility of tissue interactions, we examined cross-sectional relations among serum, tissue, and dietary lutein concentrations, anthropometric measures, and MP density in healthy adults. Significant negative correlations were found between adipose tissue lutein concentrations and MP for women, but a significant positive relation was found for men. CONCLUSION: Sex differences in lutein metabolism may be an important factor in tissue interactions and in determining MP density.

Lens formation by pigmented epithelial cell reaggregate from dorsal iris implanted into limb blastema in the adult newt.

In newt lens regeneration, the dorsal iris has lens forming ability and the ventral iris has no such capability, whereas there is no difference in the morphological criteria. To investigate the real aspects of this characteristic lens regeneration in the newt at the cellular level, a useful model system was constructed by transplanting the dorsal and ventral reaggregate derived from singly dissociated pigmented epithelial cells of the iris into the blastema of the forelimb in the newt. The lens was formed from the dorsal reaggregate with high efficiency, but not from the ventral one. No lens formation was observed in the implantation of the reaggregate into the tissue of the intact limbs. In detailed examination of the process of lens formation from the reaggregate, it was shown that tubular formation was the first step in the rearrangement of cells within the reaggregate. This was followed by depigmentation, vesicle formation with active cell growth, and the final step was lens fiber formation by transdifferentiation of epithelial cells composing the lens vesicle. The process was almost the same as in situ lens regeneration except the reconstitution of the two-layered epithelial structure was embodied as flattened tubular formation in the first step. The present study made it possible for the first time to examine lens forming ability in the reaggregate mixed with dorsal and ventral cells, because the formation of a reaggregate was started from singly dissociated cells of the dorsal and ventral cells of the iris. Mixed reaggregate experiments indicated that the existence of the dorsal cells in a cluster within the reaggregate is important in lens formation, and ventral cells showed an inhibitory effect on the formation. The present study demonstrated that the limb system thus constructed was effective for the analysis of lens formation at the cellular level and made it possible to examine the role of dorsal and ventral cells in lens regeneration.

The relationships of age changes in retinal pigment epithelium and Bruch's membrane.

PURPOSE: To study the correlations between age, Bruch's membrane (BM) thickness, retinal pigment epithelial (RPE) autofluorescence, and RPE residual body content. METHODS: Eight-millimeter-diameter macular discs from 88 unpaired human eye bank eyes were obtained within 72 hours of death, fixed in 10% neutral buffered formalin, and hemisected horizontally. One portion of the macular disc was embedded in paraffin and stained with periodic acid-Schiff for the measurement of BM thickness. RPE autofluorescence measurements were performed on unstained, deparaffinized sections. A second portion of the macular disc was prepared for electron microscopy to evaluate RPE residual body content. Linear and polynomial regression techniques were used to investigate the correlations between age, BM thickness, RPE autofluorescence, and RPE residual body content. RESULTS: Bruch's membrane thickness increased with age according to the linear model. RPE autofluorescence and RPE residual body content also increased with age, but the correlations were best approximated by a quadratic model. The correlations between RPE autofluorescence and residual body content and between BM thickness and RPE autofluorescence were best approximated by a linear regression model. There was considerable variation in these correlations between specimens and within the same age group. CONCLUSIONS: Although the changes in RPE and Bruch's membrane increased with age and there was a direct correlation between changes in the two tissues, there was considerable variation within each age group and between specimens. This probably reflects the multifactorial nature of the process.

pICln can regulate swelling-induced Cl- currents in either layer of rabbit ciliary epithelium.

Swelling-induced Cl- currents were investigated in freshly prepared non-pigmented epithelial (NPE) and pigmented epithelial (PE) cells of the rabbit ciliary body using the whole-cell patch clamp technique. Exposure of both NPE and PE cells to hypotonic stress induced Cl- currents that exhibited outward rectification and were insensitive to Ca+2. We found that swelling-induced Cl- currents in PE cell are observed shortly after isolation. The swelling-induced Cl- current showed little or no inactivation at positive membrane voltages and was sensitive to 100 microM NPPB and 100 microM DIDS. Injection of cRNA encoded rabbit pICln into Xenopus oocytes produced an outwardly rectifying Cl- current displaying features consistent with the swelling-induced Cl- current in epithelium. pICln is ubiquitous in the ciliary epithelium. It participates in the equilibration of short term tonicity alterations, a phenomenon underlying mechanisms with larger and slower amplitudes for aqueous secretion by these cells.

Melanoma maligno de coroides. Importancia de la lipofucsinosis en el diagnóstico temprano / Choroid malignant melanoma. Importance of the lipofuscin in the diagnosis

Se presenta el caso de un melanoma maligno de coroides en situación submacular, el cual se detectó en forma temprana por medio de los métodos clínicos de ecografía y fluorangiografía. Se estableció correlación anatomopatológica de estos procedimientos de gabinete. La presencia de lipofucsinas en el epitelio pigmentado de la retina se demostró en la clínica como un pigmento amarillo en la periferia de la neoplasia. Los hallazgos pronósticos concuerdan con la buena evolución de la paciente y la ausencia de matástasis.

Formoguanamine-induced inhibition of deprivation myopia in chick is accompanied by choroidal thinning while retinal function is retained.

Twenty hatchling chickens were injected intravitreally every 4 days from day 2 to day 16 with dimethyl sulphoxide (DS) in one eye and DS or formoguanamine dissolved in DS (FG.DS) with or without occlusion in the other (FG.DS.MD, DS.MD, FG.DS). At day 16, the FG.DS.MD eyes failed to show the high refractive myopia and showed less axial elongation than that developed by the DS.MD eyes. Electroretinograms indicated that at the dosage used, FG.DS does not eliminate phototransduction. Light microscopy showed choroidal and retinal thinning in DS.MD and FG.DS.MD eyes but less than in FG.DS eyes, suggesting that change in choroidal thickness is unlikely to be the primary cause of form deprivation myopia.

Development of the interphotoreceptor matrix in Xenopus laevis.

Xenopus laevis interphotoreceptor matrix (IPM) contains a relatively aqueous insoluble wheat germ agglutinin (WGA)-binding component containing unidentified sialoglycoconjugates (Wood et al [1984] J. Comp. Neurol. 228:299-307). The appearance of WGA-binding macromolecules in the IPM was assessed during late embryonic stages (32-45) and in retinal rudiment cultures, using lectin cytochemistry and Western blotting techniques. Metabolic labeling of the neural retina versus retinal pigment epithelium (RPE)-choroid of juvenile Xenopus with 35S-MET was also evaluated in vivo and in vitro. Lectin cytochemistry of eyes from developmental stages 32-42 demonstrated distinct WGA-ferritin-binding sites on the developing outer segment membranes and in the IPM compartment. At stages 44-46 extensive WGA-binding domains were present as an extracellular network with other randomly scattered domains near the retinal pigment epithelium. Retinal rudiments from stage 32-33 were isolated and allowed to differentiate in hanging drop culture (Hollyfield and Witkowsky [1974] J. Exp. Zool. 189:357-377) with or without an investing pigment epithelium. Cultures developing with RPE exhibited an elaborate IPM with an anastomosing meshwork of WGA-ferritin binding sites. In the absence of RPE only limited amounts of binding restricted to the immediate vicinity of the developing photoreceptor outer segment membranes was observed. When Western blots were probed with WGA-HRP, stage 32-45 retinas demonstrated a major WGA-binding band of 126 kD. Similar amounts of WGA-binding macromolecules were synthesized in preparations cultured in the presence or absence of the investing RPE. During development the major WGA-binding component is a 126-kD protein. Equivalent synthesis of this protein in the presence and absence of RPE suggests that the PE is not required for synthesis of this 126-kD component. These results suggest that the retina is the primary site of synthesis of the WGA-binding components of the Xenopus IPM, whereas the PE plays a principal role in their assembly and organization.

Natriuretic peptides and their receptors in human neural retina and retinal pigment epithelium.

Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) play a role in transepithelial fluid movement in the kidney and at the blood-aqueous barrier. We sought a similar natriuretic peptide-mediated regulatory system at the level of the blood-retinal barrier by investigating human neural retina and retinal pigment epithelium for the presence of ANP, BNP and their receptors. ANP and BNP binding to receptors could be demonstrated autoradiographically in all layers of the retina inclusive of the retinal pigment epithelium. Competitive preincubation with unlabeled peptides blocked the binding of the respective radioactive peptide. ANP and BNP could also be demonstrated immunohistochemically in both the neural retina and the retinal pigment epithelium. Our results suggest a role of these peptides both in the regulation of intraretinal fluid movement and--by analogy with other peptides--as possible neutrotransmittors. The localisation of ANP and BNP in the retinal pigment epithelium suggests that these peptides may influence ocular fluid homeostasis at the outer blood-retinal barrier by modulating pigment epithelial function.

Development of the eye in the North American opossum (Didelphis virginiana).

Marsupials are unique models for developmental biology-oriented research because of the immature state of their development at birth. The North American opossum (Didelphis virginiana) has several advantages over other marsupials, including large litter size, short prenatal period (12.5 d), an extended postnatal period while accessible in the pouch, and its ability to reproduce reliably in captivity. Studies of ocular development in this species have not been reported previously. The aim of the present investigation was therefore to document the major landmarks in prenatal and postnatal development of the cornea, lens, iris, ciliary body and retina. Fifteen embryos (10.5, 10.7 and 11 d postconception and 6 h after birth [12 d]) were studied by paraffin histology. Eyes of pouch young at 8 d, 2, 6, 9 and 13 wk were studied by transmission electron microscopy and light microscopy. The study revealed a similar pattern of ocular development in Didelphis to other metatherian and eutherian mammals. Differentiation of the eye is particularly rapid in the 2 d before birth. For example, although the lens vesicle separates from the surface ectoderm on d 10, by birth (2.5 d later) a primitive cornea and fused eyelids have formed, presumably to protect the eye during migration to the pouch. At birth the retinal pigment epithelium (RPE) contains melanin; however, the inner layer of the optic cup does not differentiate into an inner and outer neuroblastic layer until 8 d after birth. Around 6 wk after birth most components of the adult eye are identifiable, albeit in an immature form. These include the corneal layers, the iris (including the sphincter pupillae), ciliary processes, RPE tapetum, and a fully laminated retina with immature photoreceptors. A knowledge of the timing of major events in eye development in Didelphis and their comparison with equivalent events in human eye development should allow the appropriate choice of stages for any future experimental studies utilising this marsupial species.

The visual system of the Florida garfish, Lepisosteus platyrhincus (Ginglymodi). I. Retina.

The retina and choriocapillaris of the Florida garfish, Lepisosteus platyrhincus (Ginglymodi), was examined at the light and electron microscopic levels. The inner limiting membrane is covered by an extensive system of vitreal blood vessels emanating from the hyaloid artery, which enters the eye ventrally at the proximal end of the elongated optic nerve head. Two size classes of ganglion cell soma are segregated by optic axon fascicles and Müller cell endfeet, all of which lie at the level of the ganglion cell layer. A third class of 'displaced' ganglion cells lies at the border of the inner plexiform and inner nuclear layers amidst tightly packed amacrine, bipolar and Müller cell soma. Two layers of horizontal cells lie vitread of a synaptic zone consisting of a complex arrangement of horizontal and bipolar dendrites invaginating rod spherules and cone pedicles to form single and multiple (three to six) synaptic ribbon connections, respectively. Immediately vitread of the photoreceptor nuclei lie a population of 'displaced' bipolars. Three types of photoreceptors are characterised: unequal double cones, single cones (large and small) and rods. These show retinomotor movements where the rods elongate in the light and are masked by the pigment epithelium and contract in the dark as the pigment migrates sclerad. Ultrastructurally, 4 types of dark-staining (osmophilic) granules are described: (1) Small glycogen granules (0.033 microns) aggregated at the bases of the photoreceptor nuclei and larger similar granules (0.078 microns and termed paraboloids) vitread to the ellipsoid; (2) tapetal granules (0.32 microns) distributed throughout the dorsal four-fifths of the retinal pigment epithelium (RPE); (3) pigment granules (0.5-2.0 microns) in the RPE, concentrated in ventral retina; (4) granules or melanosomes (0.813 microns) of the choriocapillaris. The second class of granules constitute a tapetum lucidum eliciting a yellow eyeshine when viewed in the dark. Two other tapeta also exist, a guanine tapetum (irregular guanine crystals) and a tapetum fibrosum (stacks of collagen fibrils). Functional correlations are made, and the putative ancestral (primitive) condition of particular visual characters is established for the ray-finned fishes by out-group comparisons.