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Plasticity in the binocular correspondence of striate cortical receptive fields in kittens.

The influence of visual experience on the correspondence in position and orientation of receptive fields in the two eyes of cortical neurones was studied. Kittens were reared viewing the environment through lenses that magnified the image by 9% in one direction (meridional size lenses) with axes of magnification oriented 45 degrees left and right of vertical for the two eyes. The unequal deformations in the two eyes produced gradients of position disparity and systematic variation of orientation disparity which could not be influenced by eye movements. Two types of arrangement of the lenses, producing opposite disparities, were used; each was worn by two kittens. The receptive fields of cortical neurones were studied in the four kittens aged 3-4 months. In the binocular cells, the positions of the response fields were plotted, and the preferred orientations determined, using automatic stimulus variation, quantitative analysis, and eye-drift correction. By means of regression analysis, the degree of 'interocular deformation' was assessed; a coefficient D was derived from the positions, an angle beta from the orientations. D specified the position incongruity as a fraction of retinal eccentricity, beta the difference between the orientation incongruities of cells with near-vertical and near-horizontal receptive fields. Both D and beta were found to be of opposite signs in the two groups of kittens, as predicted by the optical effects of the lenses. The difference in D between the groups was 0.197 (predicted: 0.172); the difference in beta was 17.0 degrees (predicted: 18.9 degrees). Thus, the difference in visual environment had been completely compensated by adjustments in the positions as well as the orientations of the receptive fields. Since D and beta are independent of the alignment of the eyes, the differences between the groups reflected different functional connexions at the cortical level. Possible advantages of plasticity for the development of binocular vision are discussed. It is argued that the plasticity demonstrated here reflects a more general property of cortical receptive fields.

Animals↗

Encoding of binocular disparity by simple cells in the cat's visual cortex.

1. Spatiotemporal receptive fields (RFs) for left and right eyes were studied for simple cells in the cat's striate cortex to examine the idea that stereoscopic depth information is encoded via structural differences of RFs between the two eyes. Traditional models are based on neurons that possess matched RF profiles for the two eyes. We propose a model that requires a subset of simple cells with mismatched RF profiles for the two eyes in addition to those with similar RF structure. 2. A reverse correlation technique, which allows a rapid measurement of detailed RF profiles in the joint space-time domains, was used to map RFs for isolated single neurons recorded extracellularly in the anesthetized paralyzed cat. 3. Approximately 30% of our sample of cells shows substantial differences between spatial RF structure for the two eyes. Nearly all of these neurons prefer orientations between oblique and vertical, and are therefore presumed to be involved in processing horizontal disparities. On the other hand, cells that prefer orientations near horizontal have matched RF profiles for the two eyes. Considered together, these findings suggest that the visual system takes advantage of the orientation anisotropy of binocular disparities present in the retinal images. 4. For some cells, the spatial structure of the RF changes over the time course of the response (inseparable RF in the space-time domain). In these cases, the change is similar for the two eyes, and therefore the difference remains nearly constant at all times. Because the difference of the RF structure between the two eyes is the critical determinant of a cell's relative depth selectivity for the proposed model, space-time inseparability of RFs is not an obstacle for consistent representation of stereoscopic information. 5. RF parameters including amplitude, RF width, and optimal spatial frequency are generally well matched for the two eyes over the time course of the response. The preferred speed and direction of motion are also well matched for the two eyes. These results suggest that the encoding of motion in depth is not likely to be a function of simple cells in the striate cortex. 6. The results presented here are consistent with our model, in which stereoscopic depth information is encoded via differences in the spatial structure of RFs for the two eyes. This model provides a natural binocular extension of the current notion of monocular spatial form encoding by a population of simple cells. Note, however, that our findings do not exclude the possibility that positional shifts of RFs also play a role in determining the disparity selectivity of cortical neurons.

Animals↗

The effect of liquid silicone on the rabbit retina. Histologic and ultrastructural study.

Silicone oil was injected into 23 rabbit eyes, which had undergone vitrectomy and which were then examined in the electron microscope six weeks later. Retinal lesions were observed in the outer plexiform layer. They consisted mainly of a subtotal disappearance of the processes of horizontal and bipolar cells and of the synaptic terminals of the photoreceptors. The lesions were seen only in areas of the retina that had been in permanent contact with the oil bubble. They did not depend on the purity of the silicone oil injected.

Animals↗

Relationships between Müller cells and neurons in a primitive tetrapod, the Australian lungfish.

We recently proposed a model of cytogenesis which assumes that primitive ancestral mammals and premammalian vertebrates had a retinal composition that consisted of about seven neurons per Müller cell, comprising 1-2 cone photoreceptors, 1-2 rod photoreceptors, 2-3 bipolar cells, 1-2 amacrine cells, less than 1 ganglion cell, and less than 1 horizontal cell (Reichenbach & Robinson, 1995). The Australian lungfish (Neoceratodus forsteri) closely resembles the lobe-finned ancestors of land vertebrates, and has an extremely plesiomorphic nervous system. The present study, therefore, has examined the relative frequencies of retinal neurons and Müller cells (identified by immunolabelling for glutamine synthetase) in the lungfish retina. It was found that for each Müller cell there is an average of 1.9 cone photoreceptors, 1.7 rod photoreceptors, 3.1 amacrine/bipolar/horizontal cells, and 0.6 ganglion cells; amounting to a ratio of 7.3 neurons per Müller cell. These results support our conjecture that the sequence of cytogenesis in mammals is constrained by a developmental program that predates the evolution of mammals. The study also provides the first detailed morphological descriptions of lungfish Müller cells and their relationship with adjacent neurons. It was found that individual Müller cells in lungfish have a volume (more than 12,000 microns3) that is an order of magnitude higher than in mammals, yet the proportion of total retinal volume occupied by these cells (20%) is very similar.

Animals↗

Functional neuroarchitecture of the retina: hypothesis on the dysfunction of retinal dopaminergic circuitry in Parkinson's disease.

Recent morphologic and functional techniques for the study of nerve cells, such as intracellular injection and neurotransmitter immunohistochemistry, allow a new approach to the functional architecture of the retinal circuitry. Two types of dopaminergic cells are described: amacrine cells and interplexiform cells. These latter cells, which send processes to both the inner and outer plexiform layers, form a feedback loop acting at the level of horizontal cell coupling. Two molecules localized in such cells, dopamine and GABA, have antagonistic effects on horizontal cell coupling and regulate the diameter of their receptive fields which code for contrast. Changes in the ERG, VEPs and contrast sensitivity occur in Parkinsonian patients and are identical to those observed in animal models whose dopaminergic retinal system has been destroyed, thus suggesting a degenerative process of this system in Parkinson's disease. The observation of dopamine neurons, labelled by their tyrosine hydroxylase immunoreactivity, in the retina of 5 patients, led to the observation of reduced dopamine innervation in the central retina of Parkinsonian patients.

Dopamine↗

Intracellular chloride in retinal neurons: measurement and meaning.

Intracellular chloride activity measurements were obtained from mudpuppy retinal neurons using dual microelectrodes, one of which was made chloride-selective by filling the tip with chloride liquid ion exchange resin. In addition ionic substitution experiments were carried out in the perfused retina-eyecup preparation of the mudpuppy. A comparison of the membrane potential and calculated chloride equilibrium potential shows that retinal neurons differ in relative transmembrane chloride distribution. Ganglion cells have an ECl more negative than the resting membrane potential, whereas amacrine cells have a passive distribution of chloride. Horizontal cells have chloride distributed such that an increase in chloride conductance is depolarizing. On-bipolars have a chloride distribution similar to that of horizontal cells whereas off-bipolars show either passive distribution or some chloride accumulation. These findings are consistent with the idea that chloride ions may play a role as a depolarizing driving force for electrogenic activity of horizontal cells and on-bipolars. Results with Cl substitution experiments are consistent with this interpretation.

Animals↗

Neurotensin actions in the retina: mechanisms and variability.

The effects of neurotensin on mudpuppy retinal cells were studied using extracellular and intracellular electrophysiological recording techniques and bath application of the peptide. Ganglion and amacrine cells (but not bipolar or horizontal cells) were reversibly depolarized by low micromolar concentrations of neurotensin. Depolarizations also occurred with neurotensin application during cobalt block of synaptic transmission and were accompanied by decreased input resistances. This suggests neurotensin may act directly on amacrine and ganglion cells as a conventional excitatory transmitter. However, in many retinas, cells responded to light stimuli and to other drugs but not to neurotensin. These negative results are important in considering the peptide's normal role in retinal function.

Action Potentials↗

Development of the rabbit retina. I. Size of eye and retina, and postnatal cell proliferation.

Measures of rabbit eyes and retinal wholemounts were used to evaluate the development of retinal area and shape. The retina is shown to have a horizontal axis about a third longer than the vertical axis just before birth, and to adopt an almost symmetrical shape during postnatal development to adulthood. In general, retinal thickness is shown to decrease after birth, but differently in particular retinal regions: the reduction is marked in the periphery, and less pronounced in the visual streak. As an exception, the myelinated region--after it becomes really myelinated, from 9 days p.p.--even increases in thickness. In all regions of the retina, the absolute and relative thickness of the nuclear layers decreases, whereas the relative thickness of plexiform and fibrous layers increases. Proliferation of cells within the rabbit retina was studied during the first three postnatal weeks. 3H-thymidine incorporation was used to demonstrate DNA synthesis autoradiographically in histological sections as well as in enzymatically isolated retinal cells. A first proliferation phase occurs in the neuroblastic cell layer and ceases shortly after birth in the retinal center, but lasts for about one week in the retinal periphery. We found, however, a few 3H-thymidine-labeled cells as late as in the third postnatal week. These late-labeled cells were found within the nerve fiber layer and in the inner plexiform layer. The latter cells were shown to express antigens detected by antibodies directed to the intermediate-sized filament protein vimentin, which are known to label Müller cells and neuroepithelial stem cells. This was confirmed in our preparation of enzymatically isolated cells; all cells with autoradiographically labeled nuclei revealed a characteristic elongated morphology typical for Müller radial glia (and also for early neuroepithelial stem cells). 3H-thymidine-labeled cells in the nerve fiber layer were most probably astrocytic. In analogy to the brain, we conclude that the mammalian retina undergoes a series of proliferation phases: first an early phase producing both neurons and glial cells, and then a late phase producing glial cells, e.g., in the nerve fiber layer. Most probably, the late phase within the inner nuclear layer is glial as well, i.e., consists of dividing Müller cells; it cannot be excluded, however, that there may remain some mitotically active stem cells.

Animals↗

Ectopic photoreceptors and cone bipolar cells in the developing and mature retina.

An antibody against recoverin, the calcium-binding protein, labels photoreceptors, cone bipolar cells, and a subpopulation of cells in the ganglion cell layer. In the present study, we sought to establish the origin and identity of the cells expressing recoverin in the ganglion cell layer of the rat retina. By double labeling with rhodopsin, we demonstrate that early in development some of the recoverin-positive cells in the ganglion cell layer are photoreceptors. During the first postnatal week, these rhodopsin-positive cells are eliminated from the ganglion cell layer, but such neurons remain in the inner nuclear layer well into the first postnatal month. Another contingent of recoverin-positive cells, with morphological features equivalent to those of bipolar cells, is present in the postnatal retina, and approximately 50% of these neurons survive to maturity. The incidence of such cells in the ganglion cell layer was not affected by early transection of the optic nerve, a manipulation that causes rapid loss of retinal ganglion cells. These recoverin-positive cells were not double-labeled by cell-specific markers expressed by photoreceptors, rod bipolar cells, or horizontal and amacrine cells. Based on their staining with recoverin and salient morphological features, these ectopic profiles in the ganglion cell layer are most likely cone bipolar cells. Collectively, the results provide evidence for photoreceptors in the ganglion cell and inner nuclear layers of the developing retina, and a more permanent subpopulation of cone bipolar cells displaced to the ganglion cell layer.

Animals↗

Electronic simulation of cones, horizontal cells and bipolar cells of generalized vertebrate cone retina.

Electronic analogue of my theoretical model of generalized vertebrate cone retina [Siminoff: J. Theor. Biol. 86, 763 (1980)] is presented. Cone mosaic is simulated by 25 X 21 grid of phototransistors that have colored filters mounted in front of then to produce red-, green-, and blue-sensitive cones arranged in a "trichromatic" retina. Each retinal element is simulated by Summator-Integrator and unit gain voltage invertes are used to give correct polarities to output voltages. Dynamic properties of retinal elements are developed solely by temporal interplay of antagonistic input voltages with differing time courses, and spatial organization of receptive fields is developed by unit hexagons that precisely define cone input voltages to subsequent elements. Electronic model contains both color- and non-color-coded channels. Negative feedback from L-horizontal cells to cones, electrical coupling of like-cones, and electrical coupling of like-horizontal cells are simulated by feedforward circuits. Stray light is present due to light scattering properties of colored filters used to simulate color selectivety of cones. Stationary and moving spots of white and colored lights of varied sizes and intensities are used to study characteristics of electronic analogue. Results demonstrate practicality of electronic simulation to function analogues to real cone retinas to process visual stimuli and give information to higher centers as to size, shape, color and motion of objects in visual world.

Animals↗

Spatiotemporal patterns at the retinal output.

Edge enhancement in the retina is thought to be mediated by classical center-surround antagonism, first encountered as the interactions between horizontal cells and cones. But in the salamander retina these interactions do little to enhance edges. Instead, a robust dynamic interaction between amacrine and bipolar cells appears to be responsible for a sharp edge enhancement. To demonstrate this we recorded extracellularly from a single ganglion cell and moved a flashed square, 300 micro(m) on a side, over a 1.5 x 1.0 mm2 grid at 25-micro(m) increments. Playing back all of these recordings simultaneously simulated the pattern of responses that would have been measured from an array of ganglion cells. The emerging pattern of ganglion cell activity first faithfully represented the flashed square, but after approximately 60 ms the center of the representation collapsed, leaving a representation of only the edges. We inferred that the feedback synapse from amacrine to bipolar cells at gamma-aminobutyric acid-C (GABAC) receptors mediated this effect: bicuculline and strychnine were ineffective in altering the response pattern, but in picrotoxin the center of the representation did not collapse. The GABAergic amacrine cells thought to mediate this effect have quite narrow spread of processes, so the existence of this edge-enhancing effect suggests a mechanism quite different from classical lateral inhibition, namely the delayed inhibition of a spatially expanding input pattern.

Ambystoma↗

GABA(A) receptor immunoreactivity is transiently expressed in the developing outer retina.

Extensive evidence has suggested a trophic role of gamma-aminobutyric acid (GABA) on developing cone photoreceptors in postnatal retina. In a previous study, we showed that GABA raises intracellular calcium levels in the developing cones via activation of GABA(A) receptors. Using confocal microscopy in conjunction with immunocytochemistry, we have now demonstrated that (1) GABA(A) receptor subunits are localized on cone cell bodies as well as on cone pedicles, indicating that GABA has a direct, rather than indirect, effect on cones and (2) the temporal expression of GABA(A) receptor subunits coincides with the developmental effects of GABA on cone synaptogenesis. An antibody against the beta 2/3 subunits of the GABA(A) receptor and a specific cone marker peanut-agglutinin lectin (PNA) were used to double-label wholemount neonatal retinal preparations. Results show that GABA(A) receptors are transiently expressed on cone photoreceptors in the early stages of postnatal retinal development. GABA (A)receptor immunoreactivity is clearly present on cone cell bodies and their processes and on other--as yet unidentified--elements (horizontal cells?) in the outer plexiform layer. Immunoreactivity decreases within cone photoreceptor somata after postnatal day 5, but persists in the processes of the outer plexiform layer until day 7. Our results provide support for the hypothesis that GABA acts as an important developmental regulator of cone photoreceptor maturation.

Animals↗

Functional modifications in rod bipolar cells in a mouse model of retinitis pigmentosa.

The rd mouse has been widely used as an animal model of retinitis pigmentosa. In this model, a mutation of rod-specific phosphodiesterase leads to a loss of rods during the early period of postnatal life. Morphological modifications at the level of the outer plexiform layer have been shown (Proc. Nat. Acad. Sci. USA 97 (2000) 11020) in bipolar and horizontal cells. However, very little is known about the functional changes suffered by these cells postsynaptic to the degenerated rods. In the present work we have studied the neurotransmitter-induced currents in rod bipolar cells from the rd mouse retina. Currents induced by glutamate and GABA were studied by the patch clamp-whole cell technique, on rod bipolar cells enzymatically dissociated from the rd mouse retina. Data from rd animals were compared with non-dystrophic NMRI mice. GABA (30-100 micro M) and glutamate (100 micro M) were applied from a puff pipette in the near proximity of rod bipolar cell dendrites, clamped at physiological membrane potentials, and their evoked currents were studied. In rod bipolar cells from non-dystrophic mouse, puff application of glutamate induced an outward current. This current was increased twofold in absence of extracellular calcium (nominally 0 calcium). In rod bipolar cells from adult rd mouse, currents induced by glutamate were absent. Two types of GABA mediated currents were isolated in rod bipolar cells both in control and rd mouse retinas. The currents mediated by GABA(C) receptors were observed exclusively at the axon terminal, while the currents mediated by the GABA(A) receptors were observed upon GABA application to the bipolar cell dendrites. The currents mediated by GABA(A) receptors in rod bipolar cells from rd mouse were larger than those from control animals. We conclude that after the degeneration of rod photoreceptors in rd mouse, rod bipolar cells lost their glutamate (rod-neurotransmitter) input while they increase their response to GABA (horizontal cell-neurotransmitter). In our opinion, this work describes for the first time the changes in neurotransmitter sensitivity that affect rod bipolar cells after photoreceptor degeneration of the mouse retina.

Animals↗

Histogenesis of the goldfish retina.

The order of production of retinal cells was studied in embryos of the goldfish, Carassius auratus, using 3H-thymidine autoradiography. Cell division ceases first in the neuroepithelium at the fundus of the eye between embryonic stages 19 and 20 and gradually becomes restricted to the retinal margin by stage 24. In the fundus the cells whose nuclei will reside in the inner layer of the retina stop dividing earlier than the cells whose nuclei will reside in the outer retinal layers. Thus the ganglion cells in the fundus of the retina are produced first and the receptors and horizontal cells last.

Animals↗

Organization and development of horizontal cells in the goldfish retina, I: The use of monoclonal antibody AT101.

We have produced and characterized a monoclonal antibody, AT101, which selectively labels both viable and formaldehyde-fixed horizontal cell axon terminals, but not their somas or axons, of the goldfish (Carassius auratus) retina. The antigen recognized by AT101 appears to be a cell surface glycoprotein with a molecular weight of about 35,000 Daltons, and is present exclusively or predominantly in nervous tissues of all vertebrate species examined. We have used AT101 as a probe to analyze immunocytochemically the organization of horizontal cell axon terminals (HCATs) in the adult goldfish retina, and the emergence and maturation of these terminals during retinal development. Because of continued growth at the retinal margin in adult goldfish, there is a peripheral-to-central gradient in the age of cells, with the most mature in the center and the youngest in the periphery. In the center and near periphery of the adult retina, HCATs have a fusiform morphology and form a dense network in the middle and proximal part of the inner nuclear layer. In the far peripheral retina, the axon terminals appear round or ellipsoid. The retina closest to the retinal margin is devoid of AT101 staining, indicating that either HCATs are absent or the antigen recognized by AT101 is not present on HCATs at this stage. A similar sequence of changes in staining pattern is seen during development. Although AT101 staining can first be demonstrated in the larval retina at 1 month after hatching, it appears mostly as punctate structures. At a later stage, there are round or ellipsoid structures that resemble in morphology and location (in the inner nuclear layer) those found in the far peripheral adult retina. Double-labeling experiments with AT101 and antiserum against tubulin also indicate that AT101 labels the HCATs when they first appear during development. These data suggest that the emergence and maturation of HCAT is a late event in retinal development.

Animals↗

Expression of GABA in the fetal, postnatal, and adult human retinas: an immunohistochemical study.

The expression of GABA in the human fetal (12-25 weeks of gestation), postnatal (five-month-old), and adult (35-year-old) retinas was investigated by immunohistochemistry. GABA expression was seen as early as 12 weeks in the undifferentiated cells of the inner neuroblast zone; a few optic nerve fiber layer axons were clearly labeled, suggesting that some of the stained cell bodies were prospective ganglion cells, others could be displaced amacrine cells. From 16-17 to 24-25 weeks, intense labeling was found in the amacrine, displaced amacrine, and some ganglion cells. During this time period, horizontal cells (identified by calbindin immunohistochemistry), undergoing migration (periphery) and differentiation (center), expressed GABA prominently. In the postnatal retina, some horizontal cells were moderately labeled, but very weakly in a few cells, in the adult. The Müller cells developed immunoreactivity first weakly at 12 weeks and then moderately from 16-17 weeks onward. The staining was also evident in the postnatal and adult retinas, showing labeled processes of these glial cells. Virtually no axons in the adult optic nerve and nerve fiber layer were stained; the staining was restricted to a few, large ganglion cells and displaced amacrine cells: Some amacrines were also labeled. The possibility that GABA might play a role in horizontal cell differentiation and maturation is highlighted. Other evidences suggest that GABA might play a role in metabolism during retinal development.

Adult↗

pH changes in the invaginating synaptic cleft mediate feedback from horizontal cells to cone photoreceptors by modulating Ca2+ channels.

Feedback from horizontal cells (HCs) to cone photoreceptors plays a key role in the center-surround-receptive field organization of retinal neurons. Recordings from cone photoreceptors in newt retinal slices were obtained by the whole-cell patch-clamp technique, using a superfusate containing a GABA antagonist (100 microM picrotoxin). Surround illumination of the receptive field increased the voltage-dependent calcium current (ICa) in the cones, and shifted the activation voltage of ICa to negative voltages. External alkalinization also increased cone ICa and shifted its activation voltage toward negative voltages. Enrichment of the pH buffering capacity of the extracellular solution increased cone ICa, and blocked any additional increase in cone ICa by surround illumination. Hyperpolarization of the HCs by a glutamate receptor antagonist-augmented cone ICa, whereas depolarization of the HCs by kainate suppressed cone ICa. From these results, we propose the hypothesis that pH changes in the synaptic clefts, which are intimately related to the membrane voltage of the HCs, mediate the feedback from the HCs to cone photoreceptors. The feedback mediated by pH changes in the synaptic cleft may serve as an additional mechanism for the center-surround organization of the receptive field in the outer retina.

Action Potentials↗

Calcium binding protein immunoreactivity in pigeon retina.

Pigeon retina has been mapped immunocytochemically for vitamin D-dependent calcium-binding protein (D-CaBP). Immunoreactivity was found in the cones of the yellow field, but not in photoreceptors of the red field. The D-CaBP-containing cones were a subpopulation of those in the yellow field having straight fibres leading to their synaptic terminals. D-CaBP immunoreactivity was also found in horizontal cells, the amount present varying according to position along the retina, and in some amacrine cells. Immunoblots of pigeon retinal proteins separated by SDS-polyacrylamide gel electrophoresis indicated two D-CaBP forms, having apparent molecular weights of 27000 and 29000. Both these forms of D-CaBP have been found previously in rat and pigeon brain.

Animals↗