An unexpectedly low Stratus optical coherence tomography false-positive rate in the non-nasal quadrants of Asian eyes: indirect evidence of differing retinal nerve fibre layer thickness profiles according to ethnicity.

AIMS: To determine the frequency distribution of false-positive Stratus optical coherence tomography (OCT) results of healthy eyes in terms of peripapillary location. METHODS: One eye of each of 137 healthy Korean subjects was included. The false-positive rates (FPRs) relating to retinal nerve fibre layer (RNFL) thickness parameters were determined. RESULTS: Of the 12 clock-hour positions, the 2 and 4 o'clock positions showed the highest FPRs of 5.1% and 4.4%, respectively, and 7, 8 and 11 o'clock had the lowest FPRs of 0% at <5% level. At <1% level, 2 and 6 o'clock had an FPR of 0.7%, while the FPR was 0% for all other clock-hours. The FPRs for quadrant thicknesses at <5% level were: 0.7% for temporal; 0% for superior; 5.1% for nasal; and 2.2% for inferior. At <1% level, the inferior showed the highest FPR of 0.7%, and the other three quadrants 0%. According to line graph analysis, the 80-120 test point sections had FPRs of 4.4-6.6%, while the 0-40 and 210-256 sections were 0-0.7% at <5% level. At <1% level, the 40-60, 90-110 and 200-210 sections had 0.7%, 190-200 section 1.5%, and all other sections 0%. CONCLUSION: The FPR for the non-nasal region was much lower than expected at <5% level. Given that the current OCT normative database is based on a largely non-Asian population, these findings suggest that RNFL thickness profiles may differ according to ethnicity.

Normative data of outer photoreceptor layer thickness obtained by software image enhancing based on Stratus optical coherence tomography images.

AIM: To present normative data of outer photoreceptor layer thickness obtained by a new semiautomatic image analysis algorithm operating on contrast-enhanced optical coherence tomography (OCT) images. METHODS: Eight Stratus OCT3 scans from identical retinal locations from 25 normal eyes were registered and combined to form a contrast-enhanced average image. Utilising the vertical intensity gradients of the enhanced OCT images to demarcate retinal layers, thickness measurements of the outer photoreceptor- and retinal pigment epithelium layer (RPE-OS(complex)) were obtained. Additionally backscattered light within the outer nuclear layer (ONL) in the fovea was registered and compared with backscattered light within the ONL in the peripheral part of the macula (I(ratio)-ONL). RESULTS: The mean RPE-OS(complex) thickness in the foveal centre was 77.2 microm (SD = 3.95). The RPE-OS(complex) thickness in the superior macula 0.5-3 mm of the centre was significantly increased as compared with the corresponding inferior retina. In healthy subjects, the I(ratio)-ONL was 1.06. CONCLUSIONS: Contrast-enhanced OCT images enable quantification of outer photoreceptor layer thickness, and normative values may help understanding better the relationship between functional outcome and photoreceptor morphology in retinal diseases.

Energy limitation as a selective pressure on the evolution of sensory systems.

Evolution of animal morphology, physiology and behaviour is shaped by the selective pressures to which they are subject. Some selective pressures act to increase the benefits accrued whilst others act to reduce the costs incurred, affecting the cost/benefit ratio. Selective pressures therefore produce a trade-off between costs and benefits that ultimately influences the fitness of the whole organism. The nervous system has a unique position as the interface between morphology, physiology and behaviour; the final output of the nervous system is the behaviour of the animal, which is a product of both its morphology and physiology. The nervous system is under selective pressure to generate adaptive behaviour, but at the same time is subject to costs related to the amount of energy that it consumes. Characterising this trade-off between costs and benefits is essential to understanding the evolution of nervous systems, including our own. Within the nervous system, sensory systems are the most amenable to analysing costs and benefits, not only because their function can be more readily defined than that of many central brain regions and their benefits quantified in terms of their performance, but also because recent studies of sensory systems have begun to directly assess their energetic costs. Our review focuses on the visual system in particular, although the principles we discuss are equally applicable throughout the nervous system. Examples are taken from a wide range of sensory modalities in both vertebrates and invertebrates. We aim to place the studies we review into an evolutionary framework. We combine experimentally determined measures of energy consumption from whole retinas of rabbits and flies with intracellular measurements of energy consumption from single fly photoreceptors and recently constructed energy budgets for neural processing in rats to assess the contributions of various components to neuronal energy consumption. Taken together, these studies emphasize the high costs of maintaining neurons at rest and whilst signalling. A substantial proportion of neuronal energy consumption is related to the movements of ions across the neuronal cell membrane through ion channels, though other processes such as vesicle loading and transmitter recycling also consume energy. Many of the energetic costs within neurons are linked to 3Na(+)/2K(+) ATPase activity, which consumes energy to pump Na(+) and K(+) ions across the cell membrane and is essential for the maintenance of the resting potential and its restoration following signalling. Furthermore, recent studies in fly photoreceptors show that energetic costs can be related, via basic biophysical relationships, to their function. These findings emphasize that neurons are subject to a law of diminishing returns that severely penalizes excess functional capacity with increased energetic costs. The high energetic costs associated with neural tissue favour energy efficient coding and wiring schemes, which have been found in numerous sensory systems. We discuss the role of these efficient schemes in reducing the costs of information processing. Assessing evidence from a wide range of vertebrate and invertebrate examples, we show that reducing energy expenditure can account for many of the morphological features of sensory systems and has played a key role in their evolution.

Photoreceptor counting and montaging of en-face retinal images from an adaptive optics fundus camera.

A fast and efficient method for quantifying photoreceptor density in images obtained with an en-face flood-illuminated adaptive optics (AO) imaging system is described. To improve accuracy of cone counting, en-face images are analyzed over extended areas. This is achieved with two separate semiautomated algorithms: (1) a montaging algorithm that joins retinal images with overlapping common features without edge effects and (2) a cone density measurement algorithm that counts the individual cones in the montaged image. The accuracy of the cone density measurement algorithm is high, with >97% agreement for a simulated retinal image (of known density, with low contrast) and for AO images from normal eyes when compared with previously reported histological data. Our algorithms do not require spatial regularity in cone packing and are, therefore, useful for counting cones in diseased retinas, as demonstrated for eyes with Stargardt's macular dystrophy and retinitis pigmentosa.

Evolution of arthropod visual systems: development of the eyes and central visual pathways in the horseshoe crab Limulus polyphemus Linnaeus, 1758 (Chelicerata, Xiphosura).

Despite ongoing interest into the architecture, biochemistry, and physiology of the visual systems of the xiphosuran Limulus polyphemus, their ontogenetic aspects have received little attention. Thus, we explored the development of the lateral eyes and associated neuropils in late embryos and larvae of these animals. The first external evidence of the lateral eyes was the appearance of white pigment spots-guanophores associated with the rudimentary photoreceptors-on the dorsolateral side of the late embryos, suggesting that these embryos can perceive light. The first brown pigment emerges in the eyes during the last (third) embryonic molt to the trilobite stage. However, ommatidia develop from this field of pigment toward the end of the larval trilobite stage so that the young larvae at hatching do not have object recognition. Double staining with the proliferation marker bromodeoxyuridine (BrdU) and an antibody against L. polyphemus myosin III, which is concentrated in photoreceptors of this species, confirmed previous reports that, in the trilobite larvae, new cellular material is added to the eye field from an anteriorly located proliferation zone. Pulse-chase experiments indicated that these new cells differentiate into new ommatidia. Examining larval eyes labeled for opsin showed that the new ommatidia become organized into irregular rows that give the eye field a triangular appearance. Within the eye field, the ommatidia are arranged in an imperfect hexagonal array. Myosin III immunoreactivity in trilobite larvae also revealed the architecture of the central visual pathways associated with the median eye complex and the lateral eyes. Double labeling with myosin III and BrdU showed that neurogenesis persists in the larval brain and suggested that new neurons of both the lamina and the medulla originate from a single common proliferation zone. These data are compared with eye development in Drosophila melanogaster and are discussed with regard to new ideas on eye evolution in the Euarthropoda.

Polarization properties of the retinal nerve fiber layer.

Recently developed optical techniques provide quantitative structural measurements of the retinal nerve fiber layer (RNFL). A complete interpretation of these measurements requires understanding of the optical properties of the RNFL. This paper gives a review of the polarization properties and relevant anatomy of the ocular tissues, followed by a thorough discussion of the optical properties of the RNFL. The RNFL reflectance arises from light scattering from cylinders. Microtubules are a major component contributing to the reflectance. The RNFL reflectance exhibits weak intrinsic diattenuation and well preserves polarization. RNFL birefringence varies across the retina; the variation suggests that birefringence depends on the ultrastructure of the nerve fiber bundles, which offers hope that measurement of RNFL birefringence may be able to provide early detection of subcellular changes in glaucoma.

Retinal scanning laser polarimetry and methods to compensate for corneal birefringence.

Scanning laser polarimetry (SLP) was developed to provide objective assessment of the retinal nerve fiber layer (RNFL), a birefringent tissue, by measuring the total retardation in the reflected light. The birefringence of the anterior segment of the eye, mainly the cornea, is a confounding variable to the RNFL measurement. Anterior segment birefringence varies over a wide range among individuals. This paper reviews the principle of SLP and methods to measure and compensate for anterior segment birefringence as implemented in a commercial SLP system, GDx VCC. Anterior segment birefringence is measured from the macular retardance profile. It can be neutralized with a variable retarder in the GDx VCC and the measured retardance directly represents the RNFL retardance. Alternatively, a bias retarder can be introduced in the measurement beam path with approximately vertical slow axis, SLP measures the combination of the RNFL and the bias retarder, and RNFL retardance is then mathematically extracted from the measurement. The latter has the advantage of improved signal-to-noise ratio. With the combination of a visual RNFL image and rapid, objective, and reproducible assessment of the RNFL, GDx VCC provides an attractive clinical tool in glaucoma management.

In vivo thickness and birefringence determination of the human retinal nerve fiber layer using polarization-sensitive optical coherence tomography.

Thinning of the retinal nerve fiber layer and changes in retinal nerve fiber layer birefringence may both precede clinically detectable glaucomatous vision loss. We present in vivo thickness and depth-resolved birefringence measurements of the human retinal nerve fiber layer (RNFL) by use of polarization-sensitive optical coherence tomography (PS-OCT). Using a fiber-based PS-OCT setup real-time images of the human retina in vivo were recorded, co-registered with retinal video images of the location of PS-OCT scans. PS-OCT scans around the optic nerve head (ONH) of two healthy young volunteers were made using 10 concentric circles of increasing radius. Both the mean retinal nerve fiber layer thickness and mean retinal nerve fiber birefringence for each of 48 sectors on a circle were determined. The retinal nerve fiber layer thickness and birefringence varied as a function of sector around the ONH. Measured double pass phase retardation per unit depth values around the ONH range between 0.10 and 0.35 degrees/microm. The retinal nerve fiber layer becomes thinner with increasing distance from the ONH. In contrast, the birefringence does not vary significantly with increasing distance from the ONH.

The directionality of photoreceptors in the human retina.

The directional sensitivity of photoreceptors is a result of their structure that make them act as optical fibers. Therefore measurement of photoreceptor directionality is a tool for testing the physical properties of photoreceptors in vivo. Clinical studies of photoreceptor directionality are limited by the fact that psycho-physical methods for measuring the Stiles-Crawford effect are time consuming and require excellent co-operation from the subject. Thus different reflectometric techniques have been developed recently. This paper describes such methods, that allow to characterize the optical properties of photoreceptors, i.e. their orientation and directionality. Mechanisms likely to explain the discrepancy between the directionality factor values given by these techniques are discussed. Finally the functional advantages of photoreceptor optics are considered.

Orientation behavior of retinal photoreceptors in alternating electric fields.

In alternating electric (AC) fields, particles experience polarizing effects that induce dipoles that orient elongated specimens either parallel or perpendicular to the field lines. In this work we studied the behavior of photoreceptor cells' rod outer segments (ROS) in AC fields of different frequencies. We showed that at low frequencies, ROS orient parallel to the field, whereas at higher frequencies they orient perpendicular to the field lines (in the frequency range from 100 Hz to 10 MHz). We found this behavior to be dependent on the physiological state of cells (due to modifications in their electrical properties). To simulate cell damage, the membrane conductivity was changed by treating the cell with gramicidin A, which resulted in a decrease of cytosol conductivity and, consequently, in a change of the orientation behavior of the treated cells. The change of cell orientation with cytosol conductivity is rather sharp, suggesting the potential of the method for accurate evaluation of the cell physiological status. We modeled the interaction between ROS and AC fields approximating the rod cell by a prolate spheroid with a very long axis. The internal compartment of the ellipsoid was considered to be filled with an inhomogeneous medium consisting of alternating layers of membrane and cytoplasm as media modeling the disks. This theoretical model proved to be in good agreement with the experimental results and enabled the derivation (by fitting with the experimental results) of the membrane and cytosol parameters for normal and damaged cells.

Visual pigments: trading noise for fast recovery.

Spontaneous activation of rhodopsin without light absorption occurs at a much lower rate in rod photoreceptors and insect rhabdoms than in cones. The difference lies in the pigment molecules themselves, and has implications for the design of visual photoreceptors.

Photoreceptor topography and cone-specific electroretinograms.

It is implicit in many cone-specific ERG studies that the amplitude is proportional to the numbers of cones stimulated. The objective of these experiments was to test this idea by comparing ERGs obtained from different areas of the retina with histological data on cone-density distributions. The histology (Curcio et al., 1990) shows that the cumulative number of cones in the human retina increases exponentially with stimulus diameter between 0- and 40-deg eccentricity. L-, M-, and (L+M) cone-driven 30-Hz ERGs were obtained from a series of stimuli with one of the following configurations: (1) Circular stimuli of different angular subtense up to 70-deg diameter. (2) Annuli with 70-deg outer diameter but variable inner diameter. (3) Annuli of constant area but increasing eccentricity. Cone contrasts were equalized for each stimulus condition. The modulated and nonmodulated regions of the screen had the same mean hue and luminance. The data suggest that the L+M cone ERG amplitude increases with stimulus diameter in direct proportion to the estimated number of cones stimulated. Furthermore, the total L+M responses appear to be predicted from individual L and M responses by simple linear summation for both the disc and annular stimuli.

Sensory systems in amphioxus: a window on the ancestral chordate condition.

Amphioxus has an assortment of cells and organs for sensing light and mechanical stimuli. Vertebrate counterparts of these structures are not always apparent, and a strong case can be made for homology in only a few instances. For example, amphioxus has anatomically simple but plausible homologs of both the pineal and paired eyes of vertebrates. Placodal and neural crest derivatives are, however, more problematic: the evidence for an olfactory system in amphioxus is only circumstantial and, despite the variety of secondary sensory cell types that occur on the body surface in amphioxus, none are obvious homologs of vertebrate taste buds, neuromasts or acoustic hair cells. A useful perspective can nevertheless be gained by examining differences in amphioxus and vertebrate development, specifically how each specifies and positions sensory precursors, controls their proliferation, and deploys them through the body. The much larger size of vertebrate embryos and the need to cope developmentally with increased scale and cell numbers may account for some key vertebrate innovations, including placodes and neural crest. The presence or absence of specific structural adaptations, like the latter, is therefore less useful for judging homology between amphioxus and vertebrates than shared features of specific cell types. It is also clear that the duration of embryogenesis in vertebrates has been significantly extended in comparison with ancestral chordates so as to incorporate events that would originally have occurred during the post-embryonic growth period, including events of neurogenesis. Consequently, no scenario for the origin of vertebrates can be considered complete unless it deals explicitly with the whole of the life history and changes to it.

Expression of calcium transporters in the retina of the tiger salamander (Ambystoma tigrinum).

Changes in intracellular calcium concentration, [Ca2+]i, modulate the flow of visual signals across all stages of processing in the retina, yet the identities of Ca2+ transporters responsible for these changes are still largely unknown. In the current study, the distribution of plasma membrane and intracellular Ca2+ transporters in the retina of tiger salamander, a model system for physiological studies of retinal function, was determined. Plasma membrane calcium ATPases (PMCAs), responsible for high-affinity Ca2+ extrusion, were highly expressed in the salamander retina. PMCA isoforms 1, 2, and 4 were localized to photoreceptors, whereas the inner retina expressed all four isoforms. PMCA3 was expressed in a sparse population of amacrine and ganglion neurons, whereas PMCA2 was expressed in most amacrine and ganglion cells. Na+/Ca2+ exchangers, a high-capacity Ca2+ extrusion system, were expressed in the outer plexiform layer and in a subset of inner nuclear and ganglion layer cells. Intracellular Ca2+ store transporters were also represented prominently. SERCA2a, a splice variant of the sarcoplasmic-endoplasmic Ca2+ ATPase, was found mostly in photoreceptors, whereas SERCA2b was found in the majority of retinal neurons and in glial cells. The predominant endoplasmic reticulum (ER) Ca2+ channels in the salamander retina are represented by the isoform 2 of the IP3 receptor family and the isoform 2 of the ryanodine receptor family. These results indicate that Ca2+ transporters in the salamander retina are expressed in a cell type-specific manner.

Pineal organs of deep-sea fish: photopigments and structure.

We have examined the morphology and photopigments of the pineal organs from a number of mesopelagic fish, including representatives of the hatchet fish (Sternoptychidae), scaly dragon-fish (Chauliodontidae) and bristlemouths (Gonostomidae). Although these fish were caught at depths of between 500 and 1000 m, the morphological organisation of their pineal organs is remarkably similar to that of surface-dwelling fish. Photoreceptor inner and outer segments protrude into the lumen of the pineal vesicle, and the outer segment is composed of a stack of up to 20 curved disks that form a cap-like cover over the inner segment. In all species, the pineal photopigment was spectrally distinct from the retinal rod pigment, with lambdamax displaced to longer wavelengths, between approximately 485 and 503 nm. We also investigated the pineal organ of the deep demersal eel, Synaphobranchus kaupi, caught at depths below 2000 m, which possesses a rod visual pigment with lambdamax at 478 nm, but the pineal pigment has lambdamax at approximately 515 nm. In one species of hatchet fish, Argyropelecus affinis, two spectral classes of pinealocyte were identified, both spectrally distinct from the retinal rod photopigment.

Retinal morphology and electrophysiology of two caprimulgiformes birds: the cave-living and nocturnal oilbird (Steatornis caripensis), and the crepuscularly and nocturnally foraging common pauraque (Nyctidromus albicollis).

Oilbirds (Steatornis caripensis) breed in the total darkness of caves and forage at night on fruits. Common pauraques (Nyctidromus albicollis) are crepuscular and nocturnal foragers on flying insects. We examined if their retinal structure and function can be correlated with their types and periods of activity. Electroretinograms (ERGs) were obtained from anesthetized birds in photopic and scotopic conditions to a wide range of light intensities, following which the retinas were processed for histological analysis. Retinal sensitivity is higher in oilbirds than in common pauraques. Under scotopic conditions with maximum flash luminance, the average (+/- 95% CI) b-wave amplitude of oilbirds is double that of common pauraques (500.4 +/- 49.8 and 245.4 +/- 40.9 microV, respectively) but, under photopic conditions, the results are the reverse (common pauraque: 69.4 +/- 18.1; oilbird: 23.0 +/- 4.4 microV). On the other hand, the retina of both species is highly rod-dominated, but rods are highly more numerous in oilbirds than in common pauraques (rods:cones ratio: 123:1 and 5:1, respectively). In oilbirds, rods are largely thinner and their outer segments are 1.0 microm in diameter and 18.6 microm in length. They are distributed over various levels in the photoreceptor layers, an arrangement known for deep-sea fishes, but so far unknown for birds. In common pauraques, rods are patchily distributed and their outer segments are 4.0 microm in diameter and 53 microm in length. The oilbirds rod thinness allows more rods per area unit, and thus to catch more photons per area unit under darkness, while the low cone number suggests that the species has poor daytime vision, which concurs with the species cavernicolous daytime habits. The lower rod number of common pauraques, compared to oilbirds, appears counterbalanced by their patchiness and longer and thicker outer segments to provide high retinal sensitivity. In addition, common pauraques also have a tapetum. These features, combined with a higher proportion of cones, show that common pauraques are well equipped for crepuscular and nocturnal foraging on flying insects in an open environment.

The microvasculature of python pit organs: morphology and blood flow microkinetics.

Boid snakes have infrared sensing pits that resemble crotaline pits in electrophysiological function and ultrastructure, but differ in gross morphology, number, and location: boids have three or more simple pits in the labial scales vs a single facial pair with more complex morphology in the crotalines. We studied the morphology of the capillary bed and the microkinetics of blood flow in a boid snake, the ball python, Python regius, and compared them with the already known condition in crotalines. We used a Doppler blood flow recorder in conjunction with an electrocardiograph to measure blood flow and heartbeat, and resin casts, transmission electron microscopy, and laser confocal microscopy to study capillary morphology. Blood flow in response to infrared stimulus was virtually identical in the two taxa, but the morphology of the capillary bed differed drastically. In the ball python pits, the capillary bed consisted of a forest of vertically oriented loops with a characteristic dome at the top in contact with the receptor layer of the fundus. Immunohistochemical staining showed pericytes constricting the capillaries and domes with smooth muscle alpha-actin-labeled processes. Since latency of response was as short as 1 ms, the capillaries were apparently responding under local control to provide both nutrition and cooling to the heat-sensitive receptors. We concluded that mitochondria-filled receptors provided with a swiftly responding cooling system were nature's most efficient way of attaining infrared imaging.