Receptive field properties of rod-driven horizontal cells in the skate retina

The large receptive fields of retinal horizontal cells result primarily from extensive intercellular coupling via gap (electrical) junctions; thus, the extent of the receptive field provides an index of the degree to which the cells are electrically coupled. For rod-driven horizontal cells in the dark-adapted skate retina, a space constant of 1.18 +/- 0.15 mm (SD) was obtained from measurements with a moving slit stimulus, and a comparable value (1.43 +/- 0.55 mm) was obtained with variation in spot diameter. These values, and the extensive spread of a fluorescent dye (Lucifer Yellow) from the site of injection to neighboring cells, indicate that the horizontal cells of the all-rod retina of skate are well coupled electrically. Neither the receptive field properties nor the gap-junctional features of skate horizontal cells were influenced by the adaptive state of the retina: (a) the receptive field organization was unaffected by light adaptation, (b) similar dye coupling was seen in both dark- and light-adapted retinae, and (c) no significant differences were found in the gap-junctional particle densities measured in dark- and light-adapted retinas, i.e., 3,184 +/- 286/microns 2 (n = 8) and 3,073 +/- 494/microns 2 (n = 11), respectively. Moreover, the receptive fields of skate horizontal cells were not altered by either dopamine, glycine, GABA, or the GABAA receptor antagonists bicuculline and picrotoxin. We conclude that the rod-driven horizontal cells of the skate retina are tightly coupled to one another, and that the coupling is not affected by photic and pharmacological conditions that are known to modulate intercellular coupling between cone-driven horizontal cells in other species.     

Morphological types of horizontal cell in the retina of the domestic cat.

Two morphologically distinct types of horizontal cell are described from Golgi-stained whole mounts of the cat retina. They are referred to as A-type and B-type cells. The two types differ in their dendritic branching pattern, their overall size and the absence or presence of an axon. At every retinal position the dendrites of B-type cells branch more densely and overlap each other more frequently than do the dendrites of A-type cells. At equivalent retinal positions the dendritic field size of A-type cells is greater than that of B-type cells by a factor of about 1.5. Only B-type cells have an axon, which branches at the end into a large axon terminal system. The axons have no preferred direction of orientation. The stain-ability of horizontal cells by different Golgi methods is discussed.     

Dynamics of skate horizontal cells

The all-rod retina of the skate (Raja erinacea or R. oscellata) is known to have the remarkable capability of responding to incremental flashes superimposed on background intensities that initially block all light-evoked responses and are well above the level at which rods saturate in mixed rod/cone retinas. To examine further the unusual properties of the skate visual system, we have analyzed responses of their horizontal cells to intensity-modulated step, sinusoidal, and white-noise stimuli. We found that during exposures to mean intensities bright enough to block responses to incremental stimuli, decremental stimuli were also initially blocked. Thereafter, the horizontal cells underwent a slow recovery phase during which there was marked nonlinearity in their response properties. The cell first (within 2-3 min) responded to decrements in intensity and later (after greater than 10 min) became responsive to incremental stimuli. After adaptation to a steady state, however, the responses to intensity modulation were nearly linear over a broad range of modulation depths even at the brightest mean levels of illumination. Indeed, examination of the steady-state responses over a 5-log-unit range of mean intensities revealed that the amplitude of the white noise-evoked responses depended solely on contrast, and was independent of the retinal irradiance as the latter was increased from 0.02 to 20 muW/cm2; i.e., contrast sensitivity remained unchanged over this 1,000-fold increase in mean irradiance. A decrement from the mean as brief as 2 s, however, disturbed the steady state. Another unexpected finding in this all-rod retina concerns surround-enhancement, a phenomenon observed previously for cone-mediated responses of horizontal cells in the retinas of turtle and catfish. While exposure to annular illumination induced response compression and a pronounced sensitivity loss in response to incremental light flashes delivered to the dark central region, the cell's sensitivity showed a significant increase when tested with a white noise or sinusoidally modulated central spot. Unlike horizontal cells in other retinas studied thus far, however, response dynamics remained unchanged. Responses evoked either by a small spot (0.25-mm diam) or by a large field light covering the entire retina were almost identical in time course. This is in contrast with past findings from cone-driven horizontal cells whose response waveform (dynamics) was dependent upon the size of the retinal area stimulated.     

Rhodopsin photoproducts and rod sensitivity in the skate retina

The late photoproducts that result from the isomerization of rhodopsin have been identified in the isolated all-rod retina of the skate by means of rapid spectrophotometry. The sequence in which these intermediates form and decay could be described by a scheme that incorporates two pathways for the degradation of metarhodopsin II (MII) to retinol: one via metarhodopsin III (MIII) and the other (which bypasses MIII) through retinal. Computer simulation of the model yielded rate constants and spectral absorbance coefficients for the late photoproducts which fit experimental data obtained at temperatures ranging from 7 degrees C to 27 degrees C. Comparing the kinetics of the thermal reactions with the changes in rod threshold that occur during dark adaptation indicated that the decay of MII and the fall in receptor thresholds exhibit similarities with regard to their temperature dependence. However, the addition of 2 mM hydroxylamine to a perfusate bathing the retina greatly accelerated the photochemical reactions, but had no significant effect on the rate of recovery of rod sensitivity. It appears, therefore, that the late bleaching intermediates do not control the sensitivities of skate rods during dark adaptation.     

Spatial and temporal properties of luminosity horizontal cells in the turtle retina

Luminosity horizontal cells in the turtle retina respond approximately linearly to visual stimuli with contrast levels spanning a large part of the physiological range. We characterized the response properties of these cells under conditions of low photopic background illumination by measuring their spatial and temporal frequency transfer functions. Our experimental results indicate in two ways that, under these conditions, feedback from luminosity horizontal cells to cones does not play a major role in the mechanisms underlying the spatial and temporal tuning of horizontal cell responses. First, the shape of the spatial transfer function depended only weakly on the temporal frequency with which it was measured. Second, the shape of the temporal transfer function depended only weakly on the spatial frequency with which it was measured.     

Visual pigment and photoreceptor sensitivity in the isolated skate retina

Photoreceptor potentials were recorded extracellularly from the aspartate-treated, isolated retina of the skate (Raja oscellata and R. erinacea), and the effects of externally applied retinal were studied both electrophysiologically and spectrophotometrically. In the absence of applied retinal, strong light adaptation leads to an irreversible depletion of rhodopsin and a sustained elevation of receptor threshold. For example, after the bleaching of 60% of the rhodopsin initially present in dark-adapted receptors, the threshold of the receptor response stabilizes at a level about 3 log units above the dark-adapted value. The application of 11-cis retinal to strongly light-adapted photoreceptors induces both a rapid, substantial lowering of receptor threshold and a shift of the entire intensity-response curve toward greater sensitivity. Exogenous 11-cis retinal also promotes the formation of rhodopsin in bleached photoreceptors with a time-course similar to that of the sensitization measured electrophysiologically. All-trans and 13-cis retinal, when applied to strongly light-adapted receptors, fail to promote either an increase in receptor sensitivity or the formation of significant amounts of light-sensitive pigment within the receptors. However, 9-cis retinal isin. These findings provide strong evidence that the regeneration of visual pigment in the photoreceptors directly regulates the process of photochemical dark adaptation.     

Local nonuniformities in the horizontal cell syncytium of the turtle retina

Electrical coupling between horizontal cells of the turtle retina was investigated by means of two microelectrodes, stimulating and recording, inserted into neighboring cells at a fixed horizontal distance apart. Morphological coupling was estimated by studying the flow of the luminescent dye Lucifer yellow. The presence of electrical coupling was confirmed between structures of the same type (L₁ with axon terminals, L₂ withcell bodies, R/G with cells of color type) and absence of coupling between cells of different types was confirmed, although L₁ and L₂ are connected with each other directly by thin axons. The degree of electrical coupling in the syncytium of axon terminals (L₁), with a short (50 µ or less) but fixed distance between the electrodes, may vary by several times depending on the position of the microelectrodes. This local nonuniformity of coupling can be explained by the structural nonuniformity of the network of interconnected axon terminals. Local structural nonuniformities can hardly affect the functional properties of the horizontal cell syncytium under the conditions of photic stimulation of the retina.Institute of Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 17, No. 2, pp. 239–245, March–April, 1985.     

Peroxidase uptake by photoreceptor terminals of the skate retina

The photoreceptors of dark-adapted skate retinas bathed in a Ringer solution containing horseradish peroxidase (HRP) incorporate the tracer into membrane-bound compartments within the synaptic terminal of the cell; after 1 or 2 h of incubation, approx. 10-38% of the synaptic vesicles were labeled. The receptors appeared to be functioning normally throughout the incubation period, since electrical potentials of normal amplitude could be elicited in response to dimphotic stimuli. However, it was possible to block the uptake of peroxidase by a regimen of light adaptation that effectively suppressed light-induced activity in the electroretinogram. If, during incubation with peroxidase, retinas were exposed at 10-min intervals to an intense 1-ms flash from a xenon discharge tube, the receptor terminals were almost completely devoid of peroxidase; fewer than 2% of the vesicles were labeled. The suppression of HRP uptake could also be achieved in dark-adapted retinas by adding magnesium to the bathing solution, suggesting that calcium is necessary for transmitter release from vesicles in the receptor terminals. These findings are consistent with the view that vertebrate photoreceptors discharge a neurotransmitter in darkness, and that light decreases the release of this substance. It seems likely that the incorporation of peroxidase into vesicles of physiologically active receptor terminals reflects a mechanism for the retrieval of vesicle membrane after exocytosis.     

The proximal negative response and visual adaptation in the skate retina

The proximal negative response (PNR), a complex extracellular potential derived mainly from amacrine cell activity, was studied in the all-rod retina of the skate. Tetrodotoxin (10(-6) mg/ml) did not affect either the waveform or the latency of the response, indicating that the PNR reflects the graded, nonregenerative components of the amacrine cell potential. As regards its adaptive properties, the PNR exhibited both the extreme sensitivity to weak background light and the slow time course of light and dark adaptation that are characteristic of other responses from the proximal retina. Thus, the PNR, like the b-wave and ganglion cell discharge, appears to reflect adaptive processes located within the neural network of the inner retina.     

Structural properties of two types of mangrove stands on the northwestern coast of Sri Lanka

Mangrove stands in Puttalam lagoon and Dutch bay, two interconnected lagoons situated on the northwestern coast of Sri Lanka have been classified broadly into two groups, i.e., estuarine and island/mainland-fringing stands. Structural diversity of six mangrove stands, representing the two types was studied in terms of floristic composition, density, basal area, mean stand diameter, tree height, standing above-ground biomass and leaf-area index.Rhizophora mucronata andAvicennia marina were the dominant species. Higher mean stand diameters for the mangrove stands in Puttalam lagoon indicated greater maturity than the estuarine mangrove stands in Dutch bay. Nevertheless, estuarine stands in Dutch bay were structurally more complex (complexity indices 8.11–22.7) than the island/mainland-fringing mangrove stands (complexity indices 1.38–6.78). Higher number of species present in the estuarine mangrove stands is the major element that contributes to the higher values for the complexity indices for those stands. This appears to mask the contribution of stand-age to the complexity of a mangrove stand. Therefore complexity indices alone may not be used to explain adequately the structural diversity among mangrove stands.     

Structural and functional organization of the suprapatella in two cercopithecines

Gross and microscopic study of Cercopithecus aethiops pygerythrus and Papio cyanocephalus anubis shows that these cercopithecines have a quadriceps tendon the distal portion of which consists mostly of dense collagenous bundles with scattered fine elastic fibres most of which lie in the loose connective tissue planes within and around the tendon and around blood vessels. A distinct fibrovesicular structure, the suprapatella, lies within the tendon of the vastus intermedius above the pony patella. Histologically, this structure is characterised by interwoven bundles of collagenous fibres, among which are enmeshed large cells containing prominent nuclei surrounded by large clear spaces. It is postulated that this structure facilitates hyperfluxion of the knee during the initial phases of springing and jumping.     

Structural and functional properties of the non-muscle tropomyosins

Summary The non-muscle tropomyosins (TMs), isolated from such tissues as platelets, brain and thyroid, are structurally very similar to the muscle TMs, being composed of two highly -helical subunits wound around each other to form a rod-like molecule. The non-muscle TMs are shorter than the muscle TMs; sequence analysis demonstrates that each subunit of equine platelet TM consists of 247 amino acids, 37 fewer than for skeletal muscle TM. The major differences in sequence between platelet and skeletal muscle TM are found near the amino and carboxyl terminal ends of the proteins. Probably as the result of such alterations, the non-muscle TMs aggregate in a linear end-to-end manner much more weakly than do the muscle TMs. Since end-to-end interactions are responsible for the highly cooperative manner in which TM binds to actin, the non-muscle TMs have a lower affinity for actin filaments than do the muscle TMs. However, the attachment of other proteins to actin (e.g. the Tn-I subunit of skeletal muscle troponin or the S-1 subfragment of skeletal muscle myosin) can increase the affinity of actin filaments for non-muscle TM. The non-muscle TMs interact functionally with the Tn-I component of skeletal muscle troponin to inhibit the ATPase activity of muscle actomyosin and with whole troponin to regulate the muscle actomyosin ATPase in a Ca⁺⁺-dependent manner, even though one of the binding sites for troponin on skeletal TM is missing in non-muscle TM. A novel actomyosin regulatory system can be produced using Tn-I, calmodulin and non-muscle TM; in this case inhibition is released when the non-muscle TM detaches from the actin filament in the presence of Ca⁺⁺. Although it has not yet been demonstrated that the non-muscle TMs participate in a Ca⁺⁺-dependent contractile regulatory system in vivo it does appear that they are associated with actin filaments in vivo.     

Molecular cloning and functional expression of zfCx52.6: a novel connexin with hemichannel-forming properties expressed in horizontal cells of the zebrafish retina

Gap junction-mediated electrical coupling contributes to synchronous oscillatory activities of neurons, and considerable progress has been made in defining the molecular composition of gap junction channels. In particular, cloning and functional expression of gap junction proteins (connexins (Cx)) from zebrafish retina have shown that this part of the brain possesses a high degree of connexin diversity that may account for differential functional properties of electrical synapses. Here, we report the cloning and functional characterization of a new connexin, designated zebrafish Cx52.6 (zfCx52.6). This connexin shows little similarity to known connexins from fish and higher vertebrates. By combining in situ hybridization with Laser Capture Microdissection and RT-PCR, we found that this novel fish connexin is expressed in horizontal cells in the inner nuclear layer of the retina. Functional expression of zfCx52.6 in neuroblastoma cells and Xenopus oocytes led to functional gap junctional channels and, in single oocytes, induced large non-junctional membrane currents indicative of the formation of hemichannels, which were inhibited in reversible fashion by raising extracellular Ca(2+) concentrations.     

Ionic mechanism for the generation of horizontal cell potentials in isolated axolotl retina

The ionic mechanism of horizontal cell potentials was investigated in the isolated retina of the axolotl Ambystoma mexicanum. The membrane potentials of both receptors and horizontal cells were recorded intracellularly while the ionic composition of the medium flowing over the receptor side of the retina was changed. The membrane potential of the horizontal cell is highly depender side of the retina was changed. The membrane potential of the horizontal cell is highly dependent on the extracellular concentration of sodium. When the external ion concentration of either chloride or potassium was changed independently of the other, there were shifts in the membrane potential of the horizontal cell which could not be explained by changes in the equilibrium potential of these ions. If the external concentrations of both potassium and chloride ions were varied so that the product of their external concentrations did not change, the shift in the membrane potential of the horizontal cell was in the direction predicted by the Nernst equation. The results are consistent with the suggestion that in the dark the receptors release a synaptic transmitter which increases primarily the sodium conductance of the horizontal cell postsynaptic membrane.     

Reorganization of horizontal cell processes in the developing FVB/N mouse retina

We investigated the morphological changes of horizontal cells after postnatal photoreceptor degeneration in the developing FVB/N mouse retina, using immunocytochemistry with anti-calbindin D-28K. From postnatal day 14 (P14) onwards, processes emerging from horizontal cells descend into the inner plexiform layer (IPL) and ramify mainly in stratum 1 of the IPL. Electron microscopy revealed that the descending processes make synaptic contacts with bipolar cells in the outer plexiform layer. Our results clearly demonstrate that loss of photoreceptor cells induces the reorganization of horizontal cell processes in the retinas of FVB/N mice as they mature.     

White noise analysis of a chromatic type horizontal cell in the Xenopus retina

The dynamics of color-coded signal transmission in the light-adapted Xenopus retina were studied by a combination of white noise (Wiener) analysis and simultaneous recordings from two types of horizontal cells: chromatic-type horizontal cells (C-HCs) are hyperpolarized by blue light and depolarized by red light, whereas luminosity-type horizontal cells (L-HCs) are hyperpolarized by all wave-lengths. The retina was stimulated by two superimposed fields of red and blue light modulated by two independent white noise signals, and the resulting intracellular responses were decomposed into red and blue components (first-order kernels). The first-order kernels predict the intracellular responses with a small degree of error (3.5-9.5% in terms of mean square error) under conditions where modulated responses exceeded 30 mV in amplitude peak-to-peak, thus demonstrating that both red and blue modulation responses are linear. Moreover, there is little or no interaction between the red- and blue-evoked responses; i.e., nearly identical first-order kernels were obtained for one color whether the other color was modulated or not. In C-HCs (but not L-HCs), there were consistent differences in the dynamics of the red and blue responses. In the C-HC, the cutoff frequency of the red response was higher than for the blue (approximately 12 vs 5 Hz), and the red kernel was more bandpass than the blue. In the L-HC, kernel waveform and cutoff frequencies were similar for both colors (approximately 12 Hz or greater), and the time-to-peak of the L-HC kernel was always shorter than either the red or blue C-HC kernel. These results have implications for the mechanisms underlying color coding in the distal retina, and they further suggest that nonlinear phenomena, such as voltage-dependent conductances in HCs, do not contribute to the generation of modulation responses under the experimental conditions used here.     

Electrical properties of subsynaptic and nonsynaptic membranes of horizontal cells of the turtle retina

Horizontal cells of the L-type in the turtle retina were polarized by passing a steady current through extracellular electrodes. In this way controlled changes in membrane potential can be effectively produced in the region of the cell body. The hyperpolarization response of the horizontal cell to light is reversed on depolarization of the cell membrane to about the zero level. Consequently, the response of the horizontal cell to light is the result of a decrease in the EPSP, the magnitude of which remains constant in darkness. The resistance of the cell membrane depends on the membrane potential. Hyperpolarization of horizontal cells produced by bright light or by passage of a steady current was accompanied by a decrease in their membrane resistance. This nonlinearity evidently depends on the properties of the nonsynaptic membrane of the horizontal cells, whose resistance falls considerably on hyperpolarization. The results are qualitatively similar to those demonstrated previously [10] in an investigation of the horizontal cells of the fish retina.Institute for Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 5, No. 4, pp. 423–431, July–August, 1973.