Binocular interaction and steady-state visual evoked potentials

A correlate of binocular-neuron activity was found in some properties of visual evoked potentials (VEPs), such as facilitation (defined as a binocular response greater than the sum of the monocular responses) and changes in latency (shortening of binocular VEP latency as compared to that of monocular VEPs). Monocular and binocular steady-state VEPs in response to phase-alternating gratings of different contrast and both spatial and temporal frequency were recorded in three normal subjects. Fourier analysis of the responses was performed to isolate the component at the reversal frequency. Binocular VEPs showed facilitation in the low-contrast range (3%–10%). Facilitation was highest for gratings that had spatial frequency of 0.6–2 cycles/degree (c/d), alternating at 16 reversals per second. Phase shortening was found across a parameter range larger than that at which amplitude facilitation occurred. These results suggest that both amplitude facilitation and phase shortening in binocular VEPs may provide an objective measure of binocular visual function in clinical ophthalmology.Dedicated to Dr. G.K. von Norden on the occasion of his 60th birthday     

The light-induced rise in cytosolic calcium starts later than the receptor current of the Limulus ventral photoreceptor.

The intracellular arsenazo signal indicating the transient light-evoked change in cytosolic Ca2+ (or Sr2+) concentration was measured in Limulus ventral photoreceptors simultaneously with the receptor current under voltage clamp conditions at 15 degrees C. The latency of the light-evoked arsenazo response was consistently more than 25 msec longer than the latency of the electrical light response (receptor current or -potential). Replacing calcium by strontium in the superfusate caused, within 30-40 min, reversible changes: an enlargement of the arsenazo response and a considerable prolongation of both latencies, that of the electrical and that of the arsenazo response; the difference between the two latencies, however, stayed essentially constant.     

Inward rectification in Limulus ventral photoreceptors.

We examined inward rectification in Limulus ventral photoreceptors using the two-microelectrode voltage clamp. Hyperpolarization in the dark induced an inward current whose magnitude was distinctly dependent on extracellular K+ concentration, [K+0]. The [K+0] dependence resembled the characteristic [K+0] dependence of other inward rectifiers. The inward current was not dependent on extracellular Ca2+ or Na+, and it was unaffected by intracellular injection of Cl-. The hyperpolarization induced currents had two phases, an early nearly instantaneous phase and a slowly developing late phase. The currents were sensitive to extracellular barium and cesium. In voltage-pulse experiments, the magnitudes of the inwardly rectifying currents were variable from cell to cell, with some cells exhibiting negligible inward currents. Large hyperpolarizations (to membrane potentials more negative than about -140 mV) caused unstable inward current recordings, irreversible desensitization, and irreversible elevation of intracellular Ca2+ concentration. The inward rectifier provides negative feedback by tending to depolarize the cell (with inward current) in response to hyperpolarization. We suggest that the inward rectifier reduces the amount of hyperpolarization that would otherwise be generated by electrogenic processes. This feature would restrict the dynamic voltage range of the photoreceptors at very hyperpolarized potentials.     

Cone-specific mediation of rod sensitivity in trichromatic observers

PURPOSE: The slope of the rod threshold versus the illuminance (TVI) function changes with the wavelength of the background light. This study was conducted to determine whether the changes in slope are due to the stimulation of specific cone classes. METHODS: An eight-channel optical system was used to generate lights that differed in cone and rod photoreceptor illuminance. Rod flicker TVI functions were measured in normal trichromatic observers at mesopic light levels. The independent variables were (1) the relative contribution of the short (S)- and long (L)- wavelength cones to the background light (i.e., the background lights varied along S-only and L-only lines), and (2) the temporal frequency of the flickering lights (4, 7.5, and 15 Hz). RESULTS: The 4-Hz rod flicker TVI function had a slope of 0.87 when measured near W (MacLeod-Boynton chromaticity of 0.66, 1.0). At 4 and 7.5 Hz, an increase in the relative L-cone illuminance steepened the slope of the rod-only TVI curve, but an increase in the relative S-cone illuminance had no effect. The slope of the 7.5-Hz TVI function decreased at higher illuminance levels. At 15 Hz, the thresholds could be measured over only a limited range. CONCLUSIONS: The L-cone system contributes to the desensitization of the rod system at mesopic light levels, whereas, in the range of lights used in these experiments, the S-cone system apparently does not. The possibility that S-cone stimulation desensitizes the response to rod signals at higher levels of S-cone illumination cannot be eliminated.     

Light-evoked changes in extracellular calcium concentration in frog retina

Light-evoked changes in extracellular Ca2+ concentration were recorded with Ca2(+)-selective microelectrodes in the retina of the frog eyecup. A Ca-decrease at light onset and offset was found in the inner plexiform layer, and its properties are consistent with it resulting from Ca2+ influx into activated neuronal terminals. In the subretinal space, a Ca-increase at light onset and a Ca-decrease at offset were observed, and these likely arise directly from photoreceptors. A slower ON Ca-decrease was also seen here. Because it survives pharmacological isolation of the photoreceptors from post-synaptic interactions, but not physical isolation of the retina from the pigment epithelium, this component probably depends on pigment epithelial activity.     

Light adaptation and color opponency of horizontal cells in the turtle retina

Chromaticity-type (C-type) horizontal cells of the turtle retina receive antagonistic inputs from cones of different spectral types, and therefore their response to background illumination is expected to reflect light adaptation of the cones and the interactions between their antagonistic inputs. Our goal was to study the behavior of C-type horizontal cells during background illumination and to evaluate the role of wavelength in background adaptation. The photoresponses of C-type horizontal cells were recorded intracellularly in the everted eyecup preparation of the turtle Mauremys caspica during chromatic background illuminations. The voltage range of operation was either reduced or augmented, depending upon the wavelengths of the background and of the light stimuli, while the sensitivity to light was decreased by any background. The response-intensity curves were shifted to brighter intensities and became steeper as the background lights were made brighter regardless of wavelength. Comparing the effects of cone iso-luminant backgrounds on the Red/Green C-type horizontal cells indicated that background desensitization in these cells could not solely reflect background adaptation of cones but also depend upon response compression/expansion and changes in synaptic transmission. This leads to wavelength dependency of background adaptation in C-type horizontal cells, that is expressed as increased light sensitivity (smaller threshold elevation) and improved suprathreshold contrast detection when the wavelengths of the background and light stimuli were chosen to exert opponent effects on membrane potential.     

Light induced sodium dependent accumulation of calcium and potassium in the extracellular space of bee retina

Intense illumination of long duration induced a large transient increase in extracellular calcium (delta[Ca2+]o) and potassium (delta[K+]o) during and after light in bee retina when measured with ion-selective microelectrodes. Whenever a large delta[Ca2+]o appeared, it was accompanied by a transient afterdepolarization (TA). Both the increase in [Ca2+]o, [K+]o and the TA were reduced or abolished when sodium was replaced by arginine, choline or lithium (Li+) ions. At 0-Na conditions a Na independent decrease in [Ca2+]o was observed during illumination only. A pronounced transient depolarization of the photoreceptor in the dark due to transient anoxia did not result in a significant change in [Ca2+]o. In some retinae the elevated level of [K+]o after light was absent, however a small Na-dependent TA was still observed. The above findings suggest that intense long illumination induces a large Ca2+ influx into the photoreceptors which is followed by Na-dependent Ca2+ efflux due to Na-Ca exchange. The light-induced afterdepolarization arises mainly from K+ accumulation in the extracellular space but partially from the electrogenicity of Na-Ca exchange.     

Activation of electrogenic Na-Ca exchange by light in fly photoreceptors

Illumination of white-eyed Musca photoreceptors following hypoxia or the application of ruthenium red (RR, a known blocker of Ca2+ uptake into intracellular organelles) induced a transient after depolarization (TA). The TA was enhanced when external [Ca2+] was reduced; it was abolished when external [Na+] was reduced to a level that affected the receptor potential to a small degree. The TA was enhanced or depressed when the activity of Na/K pump, which controls the Na+ gradient, was enhanced or depressed respectively. This effect was observed even when the receptor potential was not affected. All of the above observations are consistent with the hypothesis that the TA is triggered by a light-induced increase in the concentration of intracellular free Ca2+ which appear to be very high, following treatments with hypoxia or RR. The high sensitivity of the TA to Na+ and Ca2+ gradients across the photoreceptors membrane strongly suggests that the TA is due to a transient activation of an electrogenic Na-Ca exchange mechanism which depolarizes the cell.     

Retinomotor movements in the frog retinal pigment epithelium: dependence of pigment migration on Na+ and Ca2+

The ionic dependence of the screening-pigment migrations in the frog retinal epithelium (RPE) was quantitatively studied with eyecups incubated in media of different compositions. Typical migrations in response to light and darkness, equivalent to those observed in the intact animal, were fully accomplished and maintained for up to 6 hr by the isolated organ bathed in Ringer solution rich with O2. Pigment migration in either direction was completed under the appropriate illumination conditions at any time during the day, indicating that circadian influences, if present in the intact animal, can be overridden in the isolated organ by light or darkness alone. Pigment aggregation toward the dark-adapted position was inhibited by: (a) low external Ca2+, (b) high external Na+, and (c) drugs expected to increase the cytoplasmic levels of either Na+, or Ca2+, like ouabain, caffeine and the ionophore A23187. However, the inhibition caused by low Ca2+ did not occur if Na+ was also reduced in the incubation medium. On the other hand, an increase in the concentration of external Ca2+ or the addition of Co2+ to the normal Ringer facilitated pigment aggregation in the dark. Pigment dispersion to the light-adapted position was unaffected by any of the above conditions. This is the first report of full and stable pigment responses in the RPE of vertebrate eyes incubated under simple physiological conditions. The results seem to conciliate a discrepancy of previous reports on the Ca2+ dependence of RPE movements, and are compatible with current views on ionic mechanisms in analogous systems of intracellular transport.     

Effect of acetylcholine on membrane potential of cultured human nonpigmented ciliary epithelial cells

Human nonpigmented ciliary epithelial cells (NPE) were grown in tissue culture after transformation with an origin-defective mutant of SV-40 DNA. In these cells membrane potentials (V) were measured using the microelectrode technique. Addition of 10(-4) M acetylcholine led to a bisphasic voltage response. An immediate, transient hyperpolarization was followed by a sustained depolarization below the steady state level. These responses were irreversibly blocked by 10(-5) M atropine. In Ca2+-free media the initial addition of acetylcholine resulted in an unchanged voltage response. A second application of acetylcholine in Ca2+-free solution evoked only an abortive response of V, and further addition had no effect on V. In the presence of Ca2+ channel blockers (10(-5) M verapamil, 1 mM Co2+) the acetylcholine-induced response of the membrane potential was not changed. The initial hyperpolarization induced by acetylcholine was reduced by 33 +/- 3% (n = 6) in the presence of 2 mM Ba2+ and by 79 +/- 6% (n = 6) in the presence of 1 mM quinidine. Moreover, the amplitude of the hyperpolarization was dependent on the extracellular K+ concentration. With increasing extracellular K+ concentration (and decreasing transmembrane K+ gradient) the acetylcholine-induced hyperpolarization was reduced. To further elucidate the role of Ca2+ in the acetylcholine-induced responses, we measured cytoplasmic Ca2+ activity using the fluorescence of intracellularly trapped Fura-2. Cytoplasmic Ca2+ activity increased immediately and transiently upon addition of acetylcholine. We conclude that acetylcholine transiently hyperpolarizes V in cultured human NPE by activation of K+ channels mediated by mobilization of Ca2+ from intracellular stores.     

Effect of calcium ion concentration on the permeability of the corneal endothelium

The effect of free Ca2+ ion concentration on the integrity of the barrier function of the corneal endothelium was studied using the endothelial permeability to carboxyfluorescein (Pac) as a quantitative index according to the method of Araie. Paired rabbit corneas were isolated and mounted in a chamber. To serve as a control, one eye of each pair was perfused with a glucose-glutathione-bicarbonate solution at a Ca2+ concentration of 1.1 mEq, the other with a solution at various Ca2+ concentrations ranging from 0.23-1.1 mEq. The Pac ratio of a solution with a Ca2+ concentration of 0.38 mEq or higher to the control solution was close to unity, and the Pac ratio of a solution with a Ca2+ concentration of 0.33 mEq or lower to the control solution was significantly greater than unity. In a separate experiment, it was found that only slight swelling was seen when a solution with a Ca2+ concentration of 0.23 mEq or higher was used; significant swelling was seen with a solution of a Ca2+ concentration of 0.17 mEq or lower. The lowest free Ca2+ concentration needed for maintaining the barrier function of the corneal endothelium, 0.38 mEq, was found to be higher than that needed for maintaining the corneal thickness, 0.23 mEq.     

Neuropeptide-induced [Ca2+]i transients in cultured bovine trabecular cells.

Various kinds of neuropeptides have been identified to be immunoreactive in the drainage angle of mammalian eyes. However, little is known about second messenger system involvement with these peptides. To determine whether some of these peptides are linked to a calcium signalling system in the trabecular meshwork (TM) cells, their effects on [Ca2+]i transients in fura-2 loaded cultured bovine TM cells were studied with a digital video-imaging system. The main findings of this study were: (1) The basal [Ca2+]i was 164.0 +/- 1.0 nM (mean +/- standard error of the mean, n = 668). (2) Of the neuropeptides examined, neuropeptide Y (NPY) (10(-6)M) is the most potent because it increased [Ca2+]i by about four-fold from the basal level. Other peptides--substance P, bombesin, calcitonin gene-related peptide, and vasoactive intestinal peptide induced smaller increases in [Ca2+]i. (3) We defined a response as positive if [Ca2+]i increased to a value that was 1.2-fold over the basal level. The majority of the TM cells reacted to NPY, whereas only 20-30% of the cells reacted to any of the other peptides. (4) The chelation of extracellular Ca2+ shortened the half-life of a NPY-induced response without affecting its latency. (5) NPY (10(-6)M) significantly increased the formation of inositol triphosphate following a 15 sec exposure. The same was the case for inositol monophosphate and inositol diphosphate. The results of this study suggest that in bovine TM cells, NPY stimulation is coupled to Ca2+ signalling through an increase in polyphosphoinositide turnover.     

The effect of adaptation on the differential sensitivity of the S-cone color system.

This paper presents a psychophysical dissection of the S-cone color system. Experiments were guided by a skeletal model that assumed a first stage consisting of S-, M- and L-cones, and a second stage of the opponent combination of the S and L+M signals. The response of the S-cone system was isolated by measuring difference thresholds between lights that were equiluminant tritanopic confusion pairs and thus differed only in S-cone excitation. Two types of mechanisms that control sensitivity in the S-cone system were identified: (i) static mechanisms that have a restricted range and thus limit discrimination to a small range of inputs; and (ii) adaptive mechanisms that change the state of the system in response to changes in steady illumination, so that the system is sensitive to small changes from the adapting light. These mechanisms were localized by lights that stimulated the S-cone system while keeping the signal constant at either the S, the L+M, or the post-opponent stage. The response function of the static mechanism was estimated by measuring difference thresholds at judgment points other than the steady adapting light. This procedure was repeated at a number of adaptation lights to examine the properties of adaptive mechanisms. The data were consistent with an elaborated model that included identical multiplicative gain control mechanisms in the S and L+M pre-opponent branches, and a post-opponent static sigmoidal nonlinearity with different amounts of compression for positive and negative opponent inputs.     

Comparison of the responses to light and to GABA of cells postsynaptic to barnacle photoreceptors (I-cells)

We tested the hypothesis that gamma-aminobutyric acid (GABA) is the transmitter released by barnacle photoreceptors onto postsynaptic cells (I-cells). GABA was applied to I-cells either by superfusion or by ejecting it with pressure from a pipette positioned close to the I-cell's soma. The I-cell's response to GABA was compared with its response to light (i.e. to the photoreceptors' transmitter) by recording intracellularly from its soma. Bath-applied (100 microns to 10 mM) and pressure-applied GABA (10 mM in pipette) hyperpolarizes I-cells by increasing their conductance, as does the photoreceptors' transmitter. The response to pressure-applied GABA consists of two components; both persist when Co2+ or Cd2+ are added to the saline to block synaptic transmission in the preparation, indicating that GABA affects the I-cell directly rather than affecting a presynaptic cell. GABA hyperpolarizes the I-cell when applied to the cell over the soma and ipsilateral arbor or over the contralateral arbor. The I-cells' responses to GABA and to light both depend on extracellular K+ and are affected by changes in intracellular and extracellular Cl-. However, picrotoxin and beta-guanidinopropionic acid block the response to pressure-applied GABA but do not block the response to light even at an order of magnitude higher concentration. Thus, GABA is not likely to be the transmitter that causes the hyperpolarizing response of the I-cell. It may be a neuromodulator or the transmitter of an unknown input to the I-cell.     

A new cornea-positive component to the ERG of the aspartate-treated frog retina?

Using isolated bullfrog retinas treated with aspartate, we have found a new cornea-positive photo-response (positive response). The positive response could be detected only when the retina was stimulated by a dim flash. The peak amplitude and the rate of initial rise of the positive response were intensity dependent. The spectral sensitivity of the positive response peaked at 500 nm. The general characteristics of the response were different from those of the PIII response. The positive response was closely related to the extracellular Na+ and Ca2+ concentration and completely abolished by 50 microM La3+. On the basis of the present findings, it was suggested that some types of calcium channels or transporters are involved in the generation of the positive response.     

Color perception under chromatic adaptation: "supersensitivity" with dim backgrounds

The amounts of 540 nm light and 660 nm light that when mixed result in a yellow (neither reddish nor greenish) percept remain in constant ratio, regardless of the total retinal illumance level of the mixture. A similar result holds if the mixture is superimposed upon a very dim 660 nm adapting field, except that a larger proportion of 540 nm light is required than in the dark (contrary to an effect of receptor desensitization, which would result in a smaller proportion of 540 nm light). Mixtures of 460 and 579 nm lights that give yellow/blue equilibrium percepts show the same effect: under very dim 579 nm adaptation, a larger proportion of 460 nm light is required than in the dark. These results cannot be accounted for by cone sensitivity reduction nor by a postreceptoral restoring effect; they reveal an additional mechanism of adaptation.     

Cell membrane stretch modulates the high-conductance Ca2+-activated K+ channel in bovine trabecular meshwork cells

PURPOSE: Anterior chamber structures are subjected to changes in intraocular pressure (IOP). Several studies have pointed out that trabecular meshwork (TM) cells are sensitive to mechanical stretch and that cell-signaling mechanisms are activated in response to elevated pressure. Because membrane stretch has been shown to be a modulator of several ionic conductances, this study was conducted to determine its effects on the high-conductance Ca(2+)-activated K(+) (BK(Ca)) channels present in TM cells. METHODS: Primary cultures of TM cells from bovine eyes were used. Patch-clamp recordings were performed in the cell-attached, inside-out, and whole-cell configurations. To stretch the cell membrane, both suction to the rear end of the patch pipette and hypotonic shock were used. Intracellular calcium concentration ([Ca(2+)](i)) was measured in TM cells loaded with fura-2, using an epifluorescence microscope coupled to a charge-coupled device (CCD) camera. RESULTS: Electrophysiological characterization of BK(Ca) channels was in agreement with previous studies. In cell-attached patches, the open probability of the BK(Ca) channel (i.e., the amount of time the channel is open) increased consistently when 14- to 45-mm Hg suctions were applied at a constant depolarized voltage. At a constant pressure (25 or 45 mm Hg), channel openings increased when depolarizing pulses were applied to the patch. Stretch activation of the BK(Ca) channel was not mediated by increases in [Ca(2+)](i), because it was present in inside-out patches maintained at a constant Ca(2+) concentration. Nevertheless, it cannot be ruled out that at low suction levels, a minimum Ca(2+) concentration is necessary for channel activation. Whole-cell currents carried by BK(Ca) channels increased when the isotonic solution in the bath was exchanged with a hypotonic solution and were selectively blocked by iberiotoxin. In our conditions, the hypotonic shock did not modify [Ca(2+)](i). CONCLUSIONS: The data show that in TM cells, open probability of the BK(Ca) channel is enhanced by membrane stretching as well as by membrane depolarization and [Ca(2+)](i). Changes in membrane tension induced by cell volume increase also activated whole-cell BK(Ca) currents. Homeostatic mechanisms in TM cells may involve BK(Ca) channel activation in response either to changes in cell volume or changes in IOP.     

A mathematical model of the regulation mechanism of Ca2+ concentration in lens in vivo

In order to elucidate the regulation mechanism of homeostasis on intracellular Ca2+ concentration in lens in vivo, the authors proposed a new mathematical model based upon the model of Ca2+ active transport. The mathematical model has new factors: Ca2+ efflux and Ca2+ influx across the cell membrane related to the difference between intracellular and extracellular Ca2+ concentration. Changes in intracellular Ca2+ concentration were investigated by numerical simulation using this model. Ca2+ efflux (active transport) is increased with intracellular Ca2+ concentration and their relationship corresponds to a Michaelis-Menten reaction. Ca2+ influx increases with the Ca2+ permeable coefficient of cell membrane. These results are consistent with the experimental results is a qualitative way and indicate that the model is suitable to elucidate the regulation mechanism of intracellular Ca2+ concentration. From the theoretical point of view, therefore, it is suggested that intracellular Ca2+ concentration may depend on two factors: one is the Ca2+ dependence of the Ca2+ active transport system, the other the Ca2+ permeability of the cell membrane.     

Effects of uncertainty and target displacement on the latency of express saccades in man

Saccadic eye movements generated in response to a gap paradigm in which the fixation light spot was extinguished 200 ms prior to presentation of the target light spot showed appreciably shorter latencies than for the overlap paradigm in which the target light spot was presented 200 ms prior to extinction of the fixation light spot. When there was unpredictability in the direction of target presentation, i.e., to the left or right of the fixation light spot, the gap paradigm evoked mainly fast regular saccades of peak latency of 155 ms with relatively few express saccades which were defined as having latencies of less than 120 ms. By contrast, when the target always appeared to the right, a substantial population of express saccades with peak latency 95 ms was now generated. There was also a change in the relationship between saccadic latency and target angular displacement which covered the range 5-35 degrees . With the overlap paradigm and unpredictability of target direction, the latencies of the slow regular saccades increased markedly with target angular displacement. This was not the case with the same target direction when the latency of slow regular, fast regular, and express saccades remained constant with increasing target angular displacement. This indicates for targets appearing in the same hemifield that the ocular motor system operates with shortest latency irrespective of target angular displacement.     

Light-induced Ca2+ release in the visible cones of the zebrafish

We used suction-pipette recording and fluo-4 fluorescence to study light-induced Ca2+ release from the visible double cones of zebrafish. In Ringer, light produces a slow decrease in fluorescence which can be fitted by the sum of two decaying exponentials with time constants of 0.5 and 3.8 s. In 0Ca2+-0Na+ solution, for which fluxes of Ca2+ across the outer segment plasma membrane are greatly reduced, light produces a slow increase in fluorescence. Both the decrease and increase are delayed after incorporation of the Ca2+ chelator BAPTA, indicating that both are produced by a change in Ca2+. If the Ca2+ pool is first released by bright light in 0Ca2+-0Na+ solution and the cone returned to Ringer, the time course of Ca2+ decline is much faster than in Ringer without previous light exposure. This indicates that the time constants of 0.5 and 3.8 s actually reflect a sum of Na+/Ca2+-K+ exchange and light-induced release of Ca2+. The Ca2+ released by light appears to come from at least two sites, the first comprising 66% of the total pool and half-released by bleaching 4.8% of the pigment. Release of the remaining Ca2+ from the second site requires the bleaching of nearly all of the pigment. If, after release, the cone is maintained in darkness, a substantial fraction of the Ca2+ returns to the release pool even in the absence of pigment regeneration. The light-induced release of Ca2+ can produce a modulation of the dark current as large as 0.75 pA independently of the normal transduction cascade, though the rise time of the current is considerably slower than the normal light response. These experiments show that Ca2+ can be released within the cone outer segment by light intensities within the physiological range of photopic vision. The role this Ca2+ release plays remains unresolved.