The objectives of this study were to (1) determine whether the glycinergic and GABAergic amacrine cells in the developing rabbit retina were neurochemically distinct at birth, (2) determine if the ratio of GABAergic to glycinergic amacrine cells was constant during development, (3) determine whether the capacity to take up a GABA analogue was restricted to GABAergic neurons, and (4) whether initiation of GABA transport into GABAergic neurons preceded the presence of a content of GABA in these neurons. We have used a novel strategy to immunolocalize a non-endogenous GABA analogue, γ-vinyl GABA, which is taken up into neurons by a GABA transporter. Examination of serial semithin resin-embedded sections of neonatal rabbit retinae that had been immunolabelled for glycine, GABA or γ-vinyl GABA revealed that at 1 day postnatum, 60% of amacrine cells contain glycine but not GABA and did not accumulate γ-vinyl GABA, which is similar to the percentage of glycinergic amacrine cells in the adult retina. The vast majority of the remaining amacrine cells contained GABA and many also transported γ-vinyl GABA; however, a significant number of GABA-containing cells failed to accumulate γ-vinyl GABA suggesting that possession of a content of GABA did not have to be preceded by, or be concomitant with, the presence of a GABA transport system. By 10 days postnatum, over 99% of GABA-containing amacrine cells also transported γ-vinyl GABA indicating their functional maturity. Analysis of the horizontal cells revealed no evidence for uptake of γ-vinyl GABA, but another GABA analogue, diaminobutyric acid, which is a substrate both for the neuron-associated GABA transporter and the glial GABA transporter, was accumulated into some horizontal cells at 21 days postnatum, a time point when these cells also contain endogenous GABA. We conclude that amacrine cells are committed to being GABAergic or glycinergic at, or prior to birth, and that in some amacrine cells, expression of a content of GABA may occur prior to the capacity to transport GABA. Conversely, in some ganglion cells transport of γ-vinyl GABA may precede a content of GABA.

The cervico-ocular reflex (COR) was investigated in the chameleon. Two kinds of responses were observed by oscillating the body (sine-wave stimuli) in the fixed-head animal: a "smooth response" of very low gain (around 0.08) and a saccadic response composed of 1–12 saccades per cycle of stimulation (depending on the stimulation frequency). Both responses were elicited in the compensatory direction (same direction as the stimulation) and exhibited a frequency dependence with low-pass properties. The saccadic response was especially developed and displayed a higher gain (up to 0.4) than the smooth response. In darkness, the saccades were triggered near the zero point (head-body alignment), whereas in the presence of a fixed visual surround they were elicited more regularly throughout the stimulation cycle. The amplitude of saccades was increased in the light. Consequently, the gain and the phase lag of the saccadic reponse were enhanced by the visual input. No visuo-cervical interaction was observed for the smooth response. Oscillating the body at a constant velocity (seesaw or ramp stimuli) revealed a frequency effect on the number of saccades (during a cycle of stimulation), but not on the gain of the response. Increasing the amplitude of oscillations augmented only very slightly the amplitude of saccades and consequently decreased the gain. Hence, the best working range of the saccadic response corresponds to body or head movements of low amplitude (up to ±20 deg) and low frequency (up to 0.25 Hz), and is improved by a visual input. These properties are discussed on a comparative point of view. It is proposed that, in chameleons, the saccadic response could contribute to gaze stabilization and add to the vestibulo-ocular and the optokinetic responses.

The rod-driven, AII amacrine cells in the mammalian retina maintain homologous gap junctions with one another as well as heterologous gap junctions with on-cone bipolar cells. We used background illumination to study whether changes in the adaptational state of the retina affected the permeabilities of these two sets of gap junctions. To access changes in permeability, we injected single AII amacrine cells with the biotinylated tracer, Neurobiotin, and measured the extent of tracer coupling to neighboring AII cells and neighboring cone bipolar cells. We also measured the center-receptive field size of All cells to assess concomitant changes in electrical coupling. Our results indicate that in well dark-adapted retinas, AII cells form relatively small networks averaging 20 amacrine cells and covering about 75 μm. The size of these networks matched closely to the size of AII cell on-center receptive fields. However, over most of their operating range, AII cells formed dramatically larger networks, averaging 326 amacrine cells, which corresponded to an increased receptive-field size. As the retina was light adapted beyond the operating range of the AII cells, they uncoupled to form networks comparable in size to those seen in well dark-adapted retinas. Our results, then, indicate that the adaptational state of the retina has a profound effect on the extent of electrical coupling between AII amacrine cells. Although we observed light-induced changes in the number of tracer-coupled cone bipolar cells, these appeared to be an epiphenomenon of changes in homologous coupling between AII amacrine cells. Therefore, in contrast to the robust changes in AII–AII coupling produced by background illumination, our data provided no evidence of a light-induced modulation of coupling between AII cells and on-cone bipolar cells.

The effect of light stimuli and prolonged darkness on the release of endogenous dopamine was measured in the white perch and hybrid bass retinas. Isolated retinas were superfused and released dopamine was measured using extraction and high-pressure liquid chromatography separation techniques. Potassium-induced release did not depend on the background illumination nor on the period of previous darkness. Steady white light did not affect release, but flickering light of 2 Hz increased release about two-fold. During prolonged darkness, the release of dopamine increased steadily over the test period of 2 h, but only if the experiments were performed at night. During the day such an increase was not observed. The increased release during prolonged darkness at night was turned off by a short period of steady white light. The release patterns obtained from the white perch and the hybrid bass were similar. However, the hybrid bass retina showed much lower levels of dopamine than did the white perch retina.

Microiontophoresis of γ-aminobutyric acid (GABA) was used to reversibly inactivate small sites of defined orientation/direction specificity in layers II-IV of cat area 17 while single cells were recorded in the same area at a horizontal distance of ~350–700 jam. We compared the effect of inactivating iso-orientation sites (where orientation preference was within 22.5 deg) and cross-orientation sites (where it differed by 45–90 deg) on orientation tuning and directionality. The influence of iso-orientation inactivation was tested in 33 cells, seven of which were subjected to alternate inactivation of two iso-orientation sites with opposite direction preference. Of the resulting 40 inactivations, only two (5%) caused significant changes in orientation tuning, whereas 26 (65%) elicited effects on directionality: namely, an increase or a decrease in response to a cell's preferred direction when its direction preference was the same as that at an inactivation site, and an increase in response to a cell's nonpreferred direction when its direction preference was opposite that at an inactivation site. It is argued that the decreases in response to the preferred direction reflected a reduction in the strength of intracortical iso-orientation excitatory connections, while the increases in response were due to the loss of iso-orientation inhibition. Of 35 cells subjected to cross-orientation inactivation, only six (17%) showed an effect on directionality, whereas 21 (60%) showed significant broadening of orientation tuning, with an increase in mean tuning width at half-height of 126%. The effects on orientation tuning were due to increases in response to nonoptimal orientations. Changes in directionality also resulted from increased responses (to preferred or nonpreferred directions) and were always accompanied by broadening of tuning. Thus, the effects of cross-orientation inactivation were presumably due to the loss of a cross-orientation inhibitory input that contributes mainly to orientation tuning by suppressing responses to nonoptimal orientations. Differential effects of iso-orientation and cross-orientation inactivation could be elicited in the same cell or in different cells from the same inactivation site. The results suggest the involvement of three different intracortical processes in the generation of orientation tuning and direction selectivity in area 17: (1) suppression of responses to nonoptimal orientations and directions as a result of cross-orientation inhibition and iso-orientation inhibition between cells with opposite direction preferences; (2) amplification of responses to optimal stimuli via iso-orientation excitatory connections; and (3) regulation of cortical amplification via iso-orientation inhibition.

Previous immunohistochemical studies combined with retrograde tracing in macaque monkeys have demonstrated that corticocortical projections can be differentiated by their content of neurofilament protein. The present study analyzed the distribution of nonphosphorylated neurofilament protein in callosally projecting neurons located at the V1/V2 border. All of the retrogradely labeled neurons were located in layer III at the V1/V2 border and at an immediately adjacent zone of area V2. A quantitative analysis showed that the vast majority (almost 95%) of these interhemispheric projection neurons contain neurofilament protein immunoreactivity. This observation differs from data obtained in other sets of callosal connections, including homotypical interhemispheric projections in the prefrontal, temporal, and parietal association cortices, that were found to contain uniformly low proportions of neurofilament protein-immunoreactive neurons. Comparably, highly variable proportions of neurofilament protein-containing neurons have been reported in intrahemispheric corticocortical pathways, including feedforward and feedback visual connections. These results indicate that neurofilament protein is a prominent neurochemical feature that identifies a particular population of interhemispheric projection neurons at the V1/V2 border, and suggest that this biochemical attribute may be critical for the function of this subset of callosal neurons.

We recorded visual evoked potentials in response to square-wave contrast-reversal checkerboards undergoing a transition in the mean contrast level. Checkerboards were modulated at 4.22 Hz (8.45-Hz reversal rate). After each set of 16 cycles of reversals, stimulus contrast abruptly switched between a “high” contrast level (0.06 to 1.0) to a “low” contrast level (0.03 to 0.5). Higher contrasts attenuated responses to lower contrasts by up to a factor of 2 during the period immediately following the contrast change. Contrast-response functions derived from the initial second following a conditioning contrast shifted by a factor of 2–4 along the contrast axis. For low-contrast stimuli, response phase was an advancing function of the contrast level in the immediately preceding second. For high-contrast stimuli, response phase was independent of the prior contrast history. Steady stimulation for periods as long as 1 min produced only minor effects on response amplitude, and no detectable effects on response phase. These observations delineate the dynamics of a contrast gain control in human vision.

I explore here whether linear mechanisms can explain directional selectivity (DS) in simple cells of the cat's striate cortex, a question suggested by a recent upswing in popularity of linear DS models. I chose a simple cell with a space-time inseparable receptive field (RF), i.e. one that shows gradually shifting latency across space, as the RF type most likely to depend on linear mechanisms of DS. However, measured responses of the cell to a moving bar were less modulated, and extended over a larger spatial region than predicted by two different popular “linear” models. They also were more DS in exhibiting a higher ratio of total spikes for the preferred direction. Each of the two models used for comparison has a single “branch” with a single spatiotemporally inseparable linear filter followed by a threshold, hence, a “1-branch” model. Nonlinear interactions between pairs of bars in a 2-bar linear superposition test of the cell also disagreed in time-course with those of the 1-branch models. The only model whose 1-bar and 2-bar predictions matched the measured cell (including a complete “4-branch” motion energy model that matches complex cells) has two branches that differ in phase by about 90 deg, i.e. in quadrature. Each branch has its own threshold that helps define the preceding spatiotemporal unit as a subunit even after the outputs of the two branches are summed. As subunit phases differ by only 90 deg, flashing bar responses of the 2-subunit model are similar to those of the 1-subunit model. Therefore, the number of subunits is hidden from view when testing with a conventional stationary bar. In summary, movement responses and nonlinear interactions between pairs of bars in the measured cell matched those of the 2-subunit model, while they disagreed with the popular 1-subunit model. Thus, multiple nonlinear subunits appear to be necessary for DS, even in simple cortical cells.

This investigation was stimulated by the historical confusion concerning the identity of certain pretectal nuclei and by large differences reported between species with respect to which nuclei receive retinal innervation. Subcortical visual nuclei were studied using immunohistochemistry to identify retinal projections labeled following intraocular injection of cholera toxin, b fragment. In addition, neuropeptide Y (NPY) or enkephalin (ENK) immunoreactive cells and fibers were also evaluated in the retinorecipient pretectal and thalamic areas. The results confirm the established view that the retina directly innervates the nucleus of the optic tract (NOT), posterior (PPT) and olivary pretectal (OPT) nuclei. However, the retina also innervates the hamster medial (MPT) and anterior (APT; dorsal division) pretectal nuclei, results not previously reported in rodents. A commissural pretectal area (CPT) sparsely innervated by retina is also described. The data show for the first time that the posterior limitans nucleus (PLi) receives a moderately dense, direct retinal input. The PLi does not project to the cortex and appears to be a pretectal, rather than thalamic, nucleus. All retinal projections are bilateral, although predominantly contralateral The PLi contains a moderately dense plexus of NPY- and ENK-IR fibers and terminals. However, peptidergic fibes also traverse the APT and connect with the dorsomedial pretectum. The OPT contains ENK- and NPY-IR neurons and fibers, but is specifically identifiable by a moderately dense plexus of ENK-IR terminals. Numerous ENK-IR neurons are found in the NOT and PPT. The latter also has moderate numbers of ENK-IR fibers and terminals b few NPY-IR neurons or fibers. The MPT contains modest numbers of ENK-IR fibers. The APT has no NPY-IR neurons or terminals, but an occasional ENK-IR neuron is seen and there is sparse ENK-IR innervation Peptidergic innervation of the visual nuclei does not appear to be derived from the retina. The results show a set of retinally innervated, contiguous nuclei extending from the thalamic ventrolateral geniculate nucleus dorsomedially to the midbrain CPT. These nuclei plus the superior colliculus comprise a dorsal “visual shell” embracing a central core of caudal thalamus and rostral midbrain.

The dendritic form of a cell may be established by many factors both intrinsic and environmental. Blockade of action potentials along the course of axons and in their postsynaptic targets dramatically alters the development of axonal morphology. The extent to which blockade of target cell activity retrogradely alters the dendritic morphology of the presynaptic cells is unknown. To determine whether the establishment of dendritic form by developing retinal canclion cells depends on activity within their targets, the sodium channel blocker, tetrodotoxin (TTX), was administered via minipumps to the diencephalon of cat fetuses from embryonic day 43 (E43) to E57. At E57 retinae were removed and living retinal ganglion cells injected in vitro with Lucifer yellow to reveal their dendritic morphology. In the TTX-treated animals both alpha and beta types of retinal ganglion cells were present, as were putative gamma cells. Overall, the dendrites of retinal ganglion cells in TTX-treated animals appeared qualitatively and Quantitatively similar to those of untreated animals. The only significant change in the TTX-treated cases was a small increase in the number of dendritic spines on the non-beta cells. These results indicate that the acquisition of basic dendritic form of developing ganglion cells is not influenced by the action potential activity within their targets, and that it is also independent of the terminal branching patterns of their axons.

Here we examine further the basis for directional selectivity (DS) in simple cells of the cat's striate cortex. We use a distinctly different input stimulus and different analysis of the output signal for the same type of space-time inseparable receptive field (RF) that was measured with flashing bars in the companion paper (Emerson, 1997). As in the companion paper, we have mimicked a popular “linear” model with a single “branch” that consists of a linear spatiotemporal filter followed by a soft threshold, which should mimic any simple cells that have a single subunit. Counterphase sinusoidal measurements of such a configuration always generate elliptically shaped polar plots of amplitude versus temporal phase, often pinched along the minor axis because of a high threshold. However, for many spatiotemporal frequencies, such polar phase plots, as measured in simple cells by others, show a consistent rotational phase skew. Here we apply counterphase analysis to the same 2-branch model as developed in the companion paper. We show that the model accounts for the skew as the summation of signals from linear filters separated spatially and temporally by approximately 90 deg (i.e. in spatiotemporal phase quadrature), each separated from the output stage by a soft-threshold nonlinearity. We also prove conclusively that such skew cannot be generated by a single-subunit configuration. This demonstration supports the proposed two-subunit structure for DS simple cells, such as in the example from the companion paper, which has strong linear contributions from its inseparable RF. The presence of at least two nonlinear subunits appears to be an obligatory concomitant of DS in all visual cortical cells. The primary function of these subunits may be to enhance the strength of responses to images moving in the preferred direction, as in complex cells. However, subunits may also aid in identifying the moving object through overcoming, at least partially, the phase-concealing properties of the neuron's threshold by generating a steady signal that effectively decreases the threshold for the preferred direction.

The GABAC receptor subtypes on bipolar cells of rats and cold-blooded vertebrates differ in their pharmacological properties and probably have different molecular compositions. With the exception of the rat, native GABAC receptors in mammals had not been studied. In ferret, whole-cell, voltage-clamp recordings were made from bipolar cells in the retinal slice preparation to determine which subtype of GABAC receptor predominated. Puff-evoked GABA currents in bipolar cells were partially reduced by the GABAA receptor antagonist bicuculline, indicating that both GABAA and GABAC receptors mediated the responses. By contrast, GABA currents of ganglion cells were always completely blocked by bicuculline, indicating that GABAA receptors predominated on these cells. Small-amplitude GABA currents of bipolar cells evoked by short-duration puffs were less sensitive to bicuculline than large-amplitude currents evoked by long-duration puffs. This indicates that GABAc receptors mediated proportionately more of the small-amplitude, puff-evoked responses and GABAA receptors mediated more of the large-amplitude, puff-evoked responses. In bipolar cells, the bicuculline-resistant component of the GABA current was entirely blocked by 3-APMPA (3-aminopropyl-(methyl)phosphonic acid), a GABAC receptor antagonist. Picrotoxin, which is relatively ineffective at rat GABAC receptors, completely blocked GABA currents in ferret bipolar cells, indicating that GABAC receptors on ferret bipolar cells resemble those in lower vertebrates rather than those in the rat retina. These results suggest that there may be a diversity of GABAc receptor subtypes on mammalian bipolar cells.

The short wavelength-sensitive (blue) cone bipolar cells was found to have a nonrandom distribution by analyzing the nearest neighbors and by calculating the density recovery profile (DRP). Blue cones had been shown previously to have a nonrandom distribution (Curcio et al., 1991). The relationship between the two arrays was then analyzed by calculating the cross-correlational density recovery profile (cDRP), which indicates the local density of blue cones around each blue cone bipolar cell. Although both cell types appeared to be distributed uniformly at the macroscopic level, the cDRP was 1.7 times higher within 15 μm of each bipolar cell perikaryon than in the surrounding area. The area of higher density was approximately the same as that in which the blue cone bipolar cells made synaptic contacts with blue cones. The finding that the blue cones and the blue cone bipolar cells were closer together than expected suggests that the positions of the perikarya of these neurons were influenced by their synaptic connections or other developmental interactions.

Morphology of the central retina and scotopic visual sensitivity were compared in juvenile albino and normally pigmented rainbow trout living under natural and reduced daylight. Outdoor albinos avoided exposing their eyes to direct sunlight, whereas normals were indifferent to it. After 4 months outdoors (Σ10,000 lux in albinos, Σ100,000 lux in normals), albinos had severely truncated or missing rod outer segments (ROS) and some missing rod ellipsoids, but normal numbers of photoreceptor nuclei and fully intact cones. Albino estimated ROS volume was only 7.1% of normal in July, but increased to 20% by the following February, mainly via an increase in numbers of ROS. However, in albinos moved indoors October 7 and exposed to 10–30 lux ambient daylight, both the number and length of ROS increased significantly, with estimated ROS volume reaching 95% of normal by 34 days. Albinos generally had more phagosomes (Σ3 X normal) and more macrophages (Σ2 X normal) in their outer retina. An optomotor reflex was used to define the effect of ROS volume on the ability to respond visually during dark adaptation. In July, albinos and normals from outdoor raceways (3 months) or indoor raceways (35 days) showed equal sensitivity after first being placed in darkness, but after 1 h in darkness, outdoor albinos with 6% of normal ROS volume were 2.0 log units less sensitive than indoor or outdoor normals, whereas indoor albinos with 53% of normal ROS volume were only 0.7 log units less sensitive. This verifies that most rod cell bodies of albino trout can persist without functional ROS in indirect sunlight, and can regrow functional outer segments in dim daylight. This finding is distinct from the extensive retinal light damage observed in albino rats exposed to more moderate cyclic light, in which entire rod cells degenerate early on.

Glutamate is the probable neurotransmitter of both retinal and cortical afferents to the cat superior colliculus (SC). The present study shows that glutamate is also contained in many postsynaptic neurons in SC. The distribution, morphology, and ultrastructure of neurons in SC were examined using glutamate antibody immunocytochemistry. Labeled cells were widely distributed throughout, but a specific laminar pattern was evident. Relatively few cells were found in the zonal and upper superficial gray layers (SGL). A dense band of intensely labeled neurons was found within the deep superficial gray and upper optic layers. Many cells were also labeled in the deeper layers. Labeled cells had varied sizes and morphologies. Soma diameters ranged from 9–67 μm, with a mean of 22 μm. Cells with stellate, vertical fusiform, and multipolar morphologies were labeled. Cells in the deep subdivision all had morphologies and sizes typical of projection neurons. To determine if labeled cells in the dense band were also projection neurons, WGA-HRP was injected into the lateral posterior nucleus and these sections were double-labeled with the glutamate antibody. Over one-half of cells in the dense band that were labeled by HRP were also obviously labeled by antibody. At the electron-microscope level, both medium- and large-sized neurons were also labeled by glutamate antibodies. These cells had different but characteristic morphologies.

In the ventral nerve photoreceptor of Limulus a short, intense flash evokes a receptor current consisting of three components. In contrast to other hypotheses, we suggested previously that only the second component of the current is activated by the phospholipase C pathway, which releases calcium from intracellular stores by inositol trisphosphate. The present paper gives further evidence to our suggestion. It is demonstrated that U-73122, a specific inhibitor for the phospholipase C, selectively blocks the second component. The first and third components are moderately affected and could still be activated after the complete block of the second one. Results support the idea that the first and third components areactivated by pathways operating independently of phospholipase C.

The ganglion cells of the primate retina include two major anatomical and functional classes: P cells which project to the four parvocellular layers of the lateral geniculate nucleus (LGN), and M cells which project to the two magnocellular layers. The characteristics of the P-cell receptive field are central to understanding early form and color vision processing (Kaplan et al., 1990; Schiller & Logothetis, 1990). In this and in the following paper, P-cell dynamics are systematically analyzed in terms of linear and nonlinear response properties. Stimuli that favor either the center or the surround of the receptive field were produced on a CRT and modulated with a broadband signal composed of multiple m-sequences (Benardete et al., 1992b; Benardete & Victor, 1994). The first-order responses were calculated and analyzed in this paper (part I). The findings are: (1) The first-order responses of the center and surround depend linearly on contrast. (2) The dynamics of the center and surround are well described by a bandpass filter model. The most significant difference between center and surround dynamics is a delay of approximately 8 ms in the surround response. (3) In the LGN, these responses are attenuated and delayed by an additional 1–5 ms. (4) The spatial transfer function of the P cell in response to drifting sine gratings at three temporal frequencies was measured. This independent method confirmed the delay between the (first-order) responses of the center and surround. This delay accounts for the dependence of the spatial transfer function on the frequency of stimulation.

The rod bipolar cell and about five types of ON cone bipolar cells depolarize to light by employing a sign-reversing metabotropic glutamate receptor. Glutamate responses are similar in both rod bipolar and cone bipolar cells, but the receptor mediating this response (mGluRo) was so far demonstrated only in rod bipolar cells. To test if ON cone bipolar cells also express mGluR6, we immunoreacted rat retina with an antibody specific for mGluRo, and studied the staining from serial ultrathin sections. We demonstrate that mGluR6 is indeed expressed in the dendritic tips of cone bipolar cells, the majority of which receive a ribbon synapse, and thus probably are ON cone bipolar cells. We further show that half of the dendritic tips contacting the cones stain for mGluR6, thus implying that all ON cone bipolar cell types express mGluR6.