Large field motion detection in goldfish, measured in the optomotor response, is based on the L-cone type, and is therefore color-blind (Schaerer & Neumeyer, 1996). In experiments using a two-choice training procedure, we investigated now whether the same holds for the detection of a small moving object (size: 8 mm diameter; velocity: 7 cm/s). In initial experiments, we found that goldfish did not discriminate between a moving and a stationary stimulus, obviously not taking attention to the cue “moving.” Therefore, random dot patterns were used in which the stimulus was visible only when moving. Using black and white random dot patterns with variable contrast between 0.2 and 1, we found that the fish could see motion only with high (0.8) contrast. In the decisive experiment, a red-green random dot pattern was used. By keeping the intensity of the red dots constant and reducing the intensity of the green dots, a narrow intensity range was found in which goldfish could no longer discriminate between the moving random dot stimulus in random dot surround and the stationary random dot pattern. The same was the case when a red moving disk was presented in green surround. This is the evidence that object motion is red-green color blind, i.e., color information cannot be used to detect the moving object. Calculations of the cone excitation values revealed that the M-cone type is decisive, as this cone type (and not the L-cone type) is not modulated by that particular red-green pattern in which the moving stimulus was invisible.

In the absence of a direct and specific marker for basket cells, the aim of this paper was to use available data to estimate the density of basket cell synaptic input to smooth and spiny neurons within layer IV of cat striate visual cortex (area 17). A linear quantitative analysis of layer IV basket cell connectivity data suggests that on average basket cells (1) comprise 25–35% of all GABAergic neurons in layer IV (3552–4736 cells mm−3), (2) account for 30–41% of all putative inhibitory dendritic synapses of layer IV spiny stellate cells (145–195 synapses cell−1) and a similar proportion of layer IV basket cells (25–37%, 71–107 synapses cell−1), and (3) provide each layer IV spiny cell with 13–45 axons and each layer IV basket cell with 6–29 axons. These estimates suggest that basket cells may be less common and provide a smaller proportion of the dendritic synaptic input to layer IV spiny and smooth neurons than previously thought. In addition, the analysis indicates that a layer IV spiny stellate cell may receive on average as many synapses and axons from layer IV basket cells as from lateral geniculate relay cells. Based on this potential numerical similarity, a geniculate-basket synaptic pairing in a spine-shaft microcircuit is hypothesized. This microcircuit could implement a type of local (dendritic) push–pull interaction underlying subfield antagonism.

To understand the generality and mechanisms of previously reported rod hue biases, we examined whether they are present for small foveal stimuli by comparing the wavelengths of the three spectral unique hues under dark-adapted and flash-bleached conditions. Rod green bias (shift of unique yellow) and rod blue bias (shift of unique green) were found for some observers with 1°-diameter foveal stimuli, the size most likely to stimulate rods. Smaller stimuli (0.2° and 0.6° diameter), which were least likely to stimulate rods, produced no large or consistent differences between dark-adapted and bleached conditions. This suggests that rod hue biases result from the local stimulation of rods by light, not from remote suppression by dark-adapted, unstimulated rods, and not from bleaching light artifacts.

We have used wild-type mice and mice possessing defects in specific retinal circuits in order to more clearly define functional circuits of the inner retina. The retina of the nob mouse lacks communication between photoreceptors and depolarizing bipolar cells (DBCs). Thus, all light driven activity in the nob mouse is mediated via remaining hyperpolarizing bipolar cell (HBC) circuits. Transducin null (Trα−/−) mice lack rod photoreceptor activity and thus remaining retinal circuits are solely generated via cone photoreceptor activity. Activation in inner retinal circuits in each of these mice was identified by monitoring light-induced expression of an immediate early gene, c-fos. The number of cells expressing c-fos in the inner retina was dependent upon stimulus intensity and was altered in a systematic fashion in mice with known retinal mutations. To determine whether c-fos is activated via circuits other than photoreceptors in the outer retina, we examined c-fos expression in tulp1−/− mice that lack photoreceptors in the outer retina; these mice showed virtually no c-fos activity following light exposure. Double-labeling immunohistochemical studies were carried out to more clearly define the population of c-fos expressing amacrine cells. Our results indicate that c-fos may be used to map functional circuits in the retina.

Previous studies of rod photoreceptors in vivo have employed a paired-flash electroretinographic (ERG) technique to determine rod response properties. To test whether absence versus presence of the ERG b-wave affects the photoreceptor response derived by the paired-flash method, we examined paired-flash-derived responses obtained from nob mice, a mutant strain with a defect in signal transduction between photoreceptors and ON bipolar cells that causes a lack of the b-wave. Normal littermates of the nob mice served as controls. The normalized amplitude-intensity relation of the derived response determined in nob mice at the near-peak time of 86 ms was similar to that determined for the controls. The full time course of the derived rod response was obtained for test flash strengths ranging from 0.11 to 17.38 scotopic cd s m−2 (sc cd s m−2). Time-course data obtained from nob and control mice exhibited significant but generally modest differences. With saturating test flash strengths, half-recovery times for the derived response of nobversus control mice differed by ∼60 ms or less about the combined (nob and control) average respective values. Time course data also were obtained before versus after intravitreal injection of l-2-amino-4-phosphonobutyrate (APB) (which blocks transmission from photoreceptors to depolarizing bipolar cells) and of cis 2,3-piperidine dicarboxylic acid (PDA) (which blocks transmission to OFF bipolar cells, and to horizontal, amacrine and ganglion cells). Neither APB nor PDA substantially affected derived responses obtained from nob or control mice. The results provide quantitative information on the effect of b-wave removal on the paired-flash-derived response in mouse. They argue against a substantial skewing effect of the b-wave on the paired-flash-derived response obtained in normal mice and are consistent with the notion that, to good approximation, this derived response represents the isolated flash response of the photoreceptors in both nob and normal mice.

The goldfish retina grows throughout the animal’s life, primarily by a balloon-like expansion. With this expansion, dendritic arbors of ganglion cells show scaled growth; arbors increase in size from small to large with no change in their architecture (Hitchcock & Easter, 1986; Bloomfield & Hitchcock, 1991). The study reported here showed that ganglion cell arbors acquire new synapses with this growth. Arbors from a single type of ganglion cell in retinas of small, young and large, old fish were intracellularly filled with horseradish peroxidase, examined electron microscopically, and the synapses contacting them counted and compared (small arbors vs. large arbors). The small and large arbors had similar numbers and orders of dendritic branches (i.e. similar architectures), but the large arbors were significantly larger than the small ones. The increase in arbor size was correlated with a 2.7x and 1.9x increase in the number of ribbon and conventional synaptic contacts, respectively. The addition of new synapses is proposed as a mechanism by which the signaling properties of the enlarging ganglion cells can remain constant.

Dopamine is a major neurotransmitter and neuromodulator in vertebrate retina. Although its pharmacological and physiological actions are well understood, the biochemical mechanisms of its signal transduction are less clear. Acting via D1 receptors, dopamine was shown to increase cyclic AMP levels in intact retina and to activate adenylate cyclase in retinal homogenates. The action via activation of D2 receptors is controversial: it was reported to decrease cyclic AMP levels in intact retina but inhibition of cyclase could not be demonstrated in retinal homogenates; also it was reported to activate rod outer segment cyclic GMP phosphodiesterase in vitro but did not decrease cyclic GMP levels in aspartate-treated retinas. We made an attempt to fractionate bovine retinal membranes and to investigate the effects of dopamine, via Dl and D2 receptors, on the synthesis and hydrolysis of cyclic AMP and cyclic GMP. Activation of cyclic AMP synthesis was noted in all fractions, but no effects were evident on cyclic nucleotide hydrolysis or cyclic GMP synthesis in any fraction. Also, D2 agonist did not inhibit cyclic AMP synthesis. These observations suggest that D2 receptors may not be directly coupled to cyclic nucleotide metabolizing enzymes in bovine retina.

The transient receptor potential channel TRPM1 is required for synaptic transmission between photoreceptors and the ON subtype of bipolar cells (ON-BPC), mediating depolarization in response to light. TRPM1 is present in the somas and postsynaptic dendritic tips of ON-BPCs. Monoclonal antibodies generated against full-length TRPM1 were found to have differential labeling patterns when used to immunostain the mouse retina, with some yielding reduced labeling of dendritic tips relative to the labeling of cell bodies. Epitope mapping revealed that those antibodies that poorly label the dendritic tips share a binding site (N2d) in the N-terminal arm near the transmembrane domain. A major splice variant of TRPM1 lacking exon 19 does not contain the N2d binding site, but quantitative immunoblotting revealed no enrichment of this variant in synaptsomes. One explanation of the differential labeling is masking of the N2d epitope by formation of a synapse-specific multiprotein complex. Identifying the binding partners that are specific for the fraction of TRPM1 present at the synapses is an ongoing challenge for understanding TRPM1 function.

Retinae of the crab Leptograpsus which had been maintained on a 12-h light/12-h dark cycle were cultured in vitro and exposed to 1 μM okadaic acid (OKA) at 0.75 h before light onset. Control retinae were subjected to the same routine and sampled at the same times without OKA treatment. At the concentration used, OKA totally inhibits types 1 and 2A protein phosphatases, minimally inhibits type 2B, and does not affect type 2C. 1 μM OKA provoked a diminution of rhabdom diameter measured at the level of the photoreceptor nuclei in the dark, some ommatidial cartridges being stripped of rhabdomeral microvilli altogether. After 1-h illumination (225–320 lux), further reduction of rhabdom diameter was modest in control retinae but precipitate in those treated with OKA. After 2 h, control rhabdom diameters showed a further, not significant, decline, but OKA had induced a resynthesis of massive structures with the light-microscopic appearance of rhabdoms. Electron microscopy revealed that they were heterogeneous and of the following kinds: (1) a minority of rhabdoms with normally disposed but distorted microvilli; (2) rhabdoms in the throes of events that parody normal assembly; and (3) rhabdomal volumes occupied by saccular organelles or by pleats or ruffles of irregular architecture. The cytoplasm of all such receptors was packed with free and bound ribosomes and endomembranes. The sequence of events parallels that seen during light-induced degeneration of photoreceptors of the Drosophila mutant w rdgBKS222. Preliminary experiments show that a protein kinase activator SC-9 mimics many of these effects in the dark in the presence of 1 μM OKA. As a working hypothesis, it is proposed that light activates protein kinases via diacylglycerols generated by the phototransduction cascade, and that in both crab retinas challenged with OKA and retinas of rdgBKS222 activation of a nuclear regulatory protein by hyperphosphorylation provokes a runaway transcription whose selectivity and extent remain to be determined.

Crayfish photoreceptors exhibit a voltage-dependent potassium conductance, GK, that is generally similar to the delayed rectifier channel described in neurons and other arthropod retinular cells. GK activation (i.e. the apparent threshold, Vth) occurs near the resting potential and GK is substantially reduced by 25 mM extracellular tetraethylammonium (TEA) and by intracellular Cs+ injections. Light exposure, sufficient to reduce visual sensitivity 100-fold, increases Vth (shifts it in the depolarizing direction) by about 20 mV. The light-dependent change in Vth does not depend upon the corresponding increase (depolarization) of the steady-state membrane potential nor does it depend upon inward calcium currents. Vth is slightly influenced by fluctuations in Ko associated with the light-elicited currents. During light exposure Ko (measured with K+-sensitive electrodes) increases by 2.1 mM (equivalent to an 8 mV increase in EK). This increase in EK makes only a modest contribution to the light-dependent change in Vth as determined by perfusion with high potassium salines. Intracellular calcium injections increase Vth by 10 to 20 mV and reduce visual sensitivity by 5- to 10-fold. The results imply that during exposure to high levels of illumination, K+ currents at the steady-state membrane potential are diminished by a calcium-dependent change in GK gating and, to a smaller degree, by a reduced K+ concentration gradient. It is notable that Ca2+ appears to inhibit both GK and the light-elicited conductance from both inside and outside the plasma membrane. As a consequence of the light-dependent change in Vth, GK makes only modest contributions to the changes in sensitivity and speed normally associated with light adaption. These functions are regulated by the transduction pathway and are revealed at the resting potential in the time course and magnitude of the light-elicited currents.

The sensitivity of rod- and cone-driven responses was studied in the isolated frog retina during the period of rapid dark adaptation following a conditioning flash which bleached a negligible amount of visual pigment. Following a conditioning flash, cone-driven b-wave responses were first enhanced and then depressed. The time courses of the enhancement and subsequent depression of cone-drive responses varies greatly with the intensity and wavelength of the conditioning flash, but were identical when the conditioning' flashes were matched for equal excitation of 502 nm rods. These changes in cone-driven response sensitivity were correlated with the desensitization and recovery of rods following the conditioning flash. When signal transmission from rods to second-order cells was interrupted by the addition of L-glutamate, the conditioning flash did not produce the above-described enhancement and subsequent depression of long-wavelength receptor potential responses. The suppression of cone-driven response therefore appears to be due to a synaptically mediated influence from 502 nm rods which is maximal when the rods are in the dark-adapted state, with little or no contribution from 433 nm rods, and no involvement of the pigment epithelium.

The hypothesis that a large, possibly toxic, increase in cellular calcium accompanies photoreceptor cell degeneration in several different Drosophila mutants was tested. The calcium content of wild type and mutant photoreceptors of Drosophila was measured using rapid freezing of the eyes and energy-dispersive x-ray analysis (e.d.x.) of cryosections and semithin sections of cryosubstituted material. Light- and dark-raised mutants of the following strains were studied: retinal degeneration B (rdgB); retinal degeneration C (rdgC); neither inactivation nor afterpotential C (ninaC), and no receptor potential A (norpA). These are light-dependent retinal degeneration mutants in which the affected gene products had been previously shown as myosin-kinase (ninaC), calcium-dependent phosphoprotein phosphatase (rdgC), phosphoinositide transfer protein (rdgB), and phospholipase C (norpA). In light-raised mutants, ommatidia of variable degrees of degeneration were observed. Mass-dense globular bodies of 200–500 nm diameter in relatively large quantities were found in the degenerating photoreceptor of all the mutants tested. These subcellular globules were found to have a very high calcium content, which was not found in wild type or in nondegenerating photoreceptors of the mutants. Nondegenerating photoreceptors were found not only in dark-raised mutants, but in smaller quantities also in light-raised mutants. Usually these globular structures contained high levels of phosphorus, indicating that at least part of the calcium in the mutant photoreceptors is precipitated as calcium phosphate. The results indicate that a large increase in cellular calcium accompanies light-induced photoreceptor degeneration in degenerating Drosophila mutants even when induced by very different mutations, suggesting that the calcium accumulation is a secondary rather than a primary effect in the degeneration process.

The ultrastructure of the pigeon and quail ventral lateral geniculate nucleus was analyzed with standard electron microscopy and horseradish peroxidase tracing of its retinal and tectal afferents. Six types of neurons were distinguished: two large, two medium-sized, and two small types.

The present study was performed as part of a systematic examination of glycine's coexistence with other classical transmitters and neuropeptides in neuronal populations of the larval tiger salamander retina. Substance P immunocytochemistry was combined with either glycine immunocytochemistry or autoradiography of glycine high-affinity uptake to examine whether tiger salamander substance P-amacrine cells express these glycine markers. Double-label analyses revealed two populations of substance P-amacrine cells that express glycine immunoreactivity and glycine high-affinity uptake. The large majority of double-labeled cells were situated in the innermost cell row of the inner nuclear layer, while a smaller number were located in the inner nuclear layer in the second cell row distal to the inner plexiform layer. Double-label immunocytochemistry revealed that these double-labeled cells accounted for 91.7% of substance P-immunoreactive amacrine cells. A slightly lower percentage (90.1%) of substance P-amacrine cells were found to exhibit a glycine high-affinity uptake mechanism. Substance P-amacrine cells that did not co-label for markers of glycine activity were situated in the innermost cell row of the inner nuclear layer. Substance P-immunoreactive displaced amacrine cells were not observed to co-label for either glycine immunoreactivity or glycine high-affinity uptake.

The present study reveals that the large majority of substance P-amacrine cells in the larval tiger salamander retina co-express markers of glycine activity. This finding suggests a functional diversity in the population of tiger salamander substance P-amacrine cells relative to their coexisting relationship with a major inhibitory neurotransmitter.

Light decrements are mediated by two distinct groups of rod pathways in the dark-adapted retina that can be differentiated on the basis of their sensitivity to the glutamate agonist DL-2-amino-phosphonobutyric (APB). By means of the APB sensitive pathway, rods transmit light decrements via rod bipolar cells to AII amacrine cells, then to Off cone bipolar cells, which in turn innervate the dendrites of Off ganglion cells. APB hyperpolarizes rod bipolar cells, thus blocking this rod pathway. With APB insensitive pathways, rods either directly synapse onto Off cone bipolar cells, or rods pass light decrement signal to cones by gap junctions. In the present study, whole-cell patch-clamp recordings were made from ganglion cells in the dark-adapted mouse retina to investigate the functional properties of APB sensitive and insensitive rod pathways. The results revealed several clear-cut differences between the APB sensitive and APB insensitive rod pathways. The latency of Off responses to a flashing spot of light was significantly shorter for the APB insensitive pathways than those for the APB sensitive pathway. Moreover, Off responses of the APB insensitive pathways were found to be capable of following substantially higher stimulus frequencies. Nitric oxide was found to selectively block Off responses in the APB sensitive rod pathway. Collectively, these results provide evidence that the APB sensitive and insensitive rod pathways can convey different types of information signaling light decrements in the dark-adapted retina.

Neurones activated through the corpus callosum (CC) in the cat visual cortex are known to be almost entirely located at the 17/18 border. They are orientation selective and display receptive fields (RFs) distributed along the central vertical meridian of the visual field (“visual midline”). Most of these cells are binocular, and many of them are activated both from the contralateral eye through the CC, and from the ipsilateral eye via the direct retino-geniculo-cortical (GC) pathway. These two pathways do not carry exactly the same information, leading to interocular disparity between pairs of RFs along the visual midline. Recently, we have demonstrated that a few weeks of unilateral paralytic strabismus surgically induced at adulthood does not alter the cortical distribution of these units but leads to a loss of their orientation selectivity and an increase of their RF size, mainly toward the ipsilateral hemifield when transcallosally activated (Watroba et al., 2001). To investigate interocular disparity, here we compared these RF changes to those occurring in the same neurones when activated through the ipsilateral direct GC route. The 17/18 transition zone and the bordering medial region within A17 were distinguished, as they display different interhemispheric connectivity. In these strabismics, some changes were noticed, but were basically identical in both recording zones. Ocular dominance was not altered, nor was the spatial distribution of the RFs with respect to the visual midline, nor the amplitude of position disparity between pairs of RFs. On the other hand, strabismus induced a loss of orientation selectivity regardless of whether neurones were activated directly or through the CC. Both types of RFs also widened, but in opposite directions with respect to the visual midline. This led to changes in incidences of the different types of position disparity. The overlap between pairs of RFs also increased. Based on these differences, we suggest that the contribution of the CC to binocular vision along the midline in the adult might be modulated through several intrinsic cortical mechanisms.

Omitting a stimulus from a train of repetitive stimuli, by either interrupting or terminating the train, can elicit an electrophysiological response that occurs at the time appropriate for the omitted stimulus. This study investigated whether such an omitted stimulus response (OSR) is present in the flicker electroretinogram (ERG) of the human cone system. ERGs were recorded from 11 visually normal subjects in response to full-field sinusoidal flicker trains presented against a rod-desensitizing adapting field at frequencies ranging from 12.5 to 100 Hz. Recordings were synchronized with the onset of the stimulus trains, and the amplitude and relative delay of any additional ERG responses following the offset of the flicker train were analyzed. At stimulus frequencies below 35 Hz, the number of ERG responses always equaled the number of stimulus cycles. However, over the frequency range of 38.5 to 100 Hz, the ERG contained an extra response following flicker train offset. At stimulus frequencies from 38.5 to 62.5 Hz, there was a constant delay between the peak of the extra ERG response and the time at which the next stimulus would have occurred had the flicker train continued. This constant delay is characteristic of an OSR. In addition, an extra ERG response was apparent at these same stimulus frequencies if the flicker train was interrupted by omitting stimulus cycles from the middle of the train. The pattern of ERG findings is consistent with a recently proposed model of the OSR that attributes the phenomenon to a resonant oscillation in retinal bipolar cells.

We have investigated by immunofluorescence the appearance of immunoreactive guanosine 3′-5′ cyclic monophosphate phosphodiesterase (cGMP-PDE) during the postnatal development of the retina of the pigmented rat. We show that a sudden increase in immunoreactivity takes place during postnatal day five (P5), when rod outer segments begin to form; immunoreactivity develops rapidly in the following days. Labeling is restricted to the developing photoreceptor outer segments, sparing other retinal cells, as confirmed by electron microscopy immunocytochemistry. In addition, cGMP-PDE immunoreactivity follows a center-to-periphery gradient paralleling photoreceptor differentiation. It appears that cGMP-PDE is expressed when the photoreceptor subcellular compartments are already formed, and represents a specific marker of late photoreceptor differentiation. The appearance of cGMP-PDE during development is temporally correlated with the appearance of other proteins of the phototransduction machinery.

Even during active fixation, small eye movements persist that might be expected to interfere with vision. Numerous brain mechanisms probably contribute to discounting this jitter. Changes in the timing of responses in the visual thalamus associated with fixational saccades are considered in this study. Activity of single neurons in alert monkey lateral geniculate nucleus (LGN) was recorded during fixation while pseudorandom visual noise stimuli were presented. The position of the stimulus on the display monitor was adjusted based on eye position measurements to control for changes in retinal locations due to eye movements. A method for extracting nonstationary first-order response mechanisms was applied, so that changes around the times of saccades could be observed. Saccade-related changes were seen in both amplitude and timing of geniculate responses. Amplitudes were greatly reduced around saccades. Timing was retarded slightly during a window of about 200 ms around saccades. That is, responses became more sustained. These effects were found in both parvocellular and magnocellular neurons. Timing changes in LGN might play a role in maintaining cortical responses to visual stimuli in the presence of eye movements, compensating for the spatial shifts caused by saccades via these shifts in timing.