Early retinal studies categorized ganglion cell behavior as either linear or nonlinear and rectifying as represented by the familiar X- and Y-type ganglion cells in cat. Nonlinear behavior is in large part a consequence of the rectifying nonlinearities inherent in synaptic transmission. These nonlinear signals underlie many special functions in retinal processing, including motion detection, motion in motion, and local edge detection. But linear behavior is also required for some visual processing tasks. For these tasks, the inherently nonlinear signals are “linearized” by “crossover inhibition.” Linearization utilizes a circuitry whereby nonlinear ON inhibition adds with nonlinear OFF excitation or ON excitation adds with OFF inhibition to generate a more linear postsynaptic voltage response. Crossover inhibition has now been measured in most bipolar, amacrine, and ganglion cells. Functionally crossover inhibition enhances edge detection, allows ganglion cells to recognize luminance-neutral patterns with their receptive fields, permits ganglion cells to distinguish contrast from luminance, and maintains a more constant conductance during the light response. In some cases, crossover extends the operating range of cone-driven OFF ganglion cells into the scotopic levels. Crossover inhibition is also found in neurons of the lateral geniculate nucleus and V1.

Full-field electroretinograms were recorded from five normal human subjects using white light (mean luminance: 250 cd/m2) sine wave stimuli at different frequencies and contrasts. In agreement with previous studies, we found that the amplitude of the fundamental component displayed a dip at about 12 Hz, coinciding with a maximum in the second harmonic component, indicating frequency doubling of the responses. By including measurements at different contrasts, we were able to recognize two (sine-like and transient) response components. We found that the waveform of the transient response was relatively frequency independent. An algorithm to separate the two components was developed. The interaction between these two components can explain the frequency-doubled responses around 12 Hz. The sine-like component is more linear and prominent in the low-frequency region, whereas the transient seems to be more nonlinear and prominent in the high-frequency region.

Some evidence suggests that retinal pigment epithelium (RPE) can express nicotinic acetylcholine receptors (nAChRs) as described for other epithelial cells, where nAChRs have been involved in processes such as cell development, cell death, cell migration, and angiogenesis. This study is designed to determine the expression and activity of α7 nAChRs in RPE cells. Reverse transcriptase (RT)-PCR was performed to test the expression of nicotinic α7 subunit in bovine RPE cells. Protein expression was determined by Western blot and by immunocytochemistry. Expression of nicotinic α7 subunits was also analyzed in cryostat sections of albino rat retina. Changes in protein expression were tested under hypoxic conditions. Functional nAChRs were studied by examining the Ca2+ transients elicited by nicotine and acetylcholine stimulation in fura-2–loaded cells. Expression of endogenous modulators of nAChRs was analyzed by RT-PCR and Western blot in retina and RPE. Cultured bovine RPE cells expressed nicotinic receptors containing α7 subunit. RT-PCR amplified the expected specific α7 fragment. Western blotting showed expression at the protein level, with a specific band being found at 57 kDa in both cultured and freshly isolated RPE cells. Expression of nAChRs was confirmed for cultured cells by immunofluorescence. Immunohistochemistry confirmed α7 receptor expression in rat RPE retina. α7 receptor expression was down-regulated by long-term hypoxia. A small subpopulation of RPE cultured cells showed functional nAChRs, as evidenced by the selective response elicited by nicotine and acetylcholine stimulation. Expression of the endogenous nicotinic receptors’ modulator lynx1 was confirmed in bovine retina and RPE, and expression of lynx1 and other endogenous nicotinic receptor modulators (SLURP1 and RGD1308195) were also confirmed in rat retina. These results suggest that nAChRs could have a significant role in RPE, which may not be related to the traditional role in nerve transmission but could more likely be related to the nonneuronal cholinergic system in the eye.

Adenosine-to-inosine (A-to-I) RNA editing is a cotranscriptional or posttranscriptional gene regulatory mechanism that increases the diversity of the proteome in the nervous system. Recently, the transcript for GABA type A receptor subunit α3 was found to be subjected to RNA editing. The aim of this study was to determine if editing of the chicken α3 subunit transcript occurs in the retina and if the editing is temporally regulated during development. We also raised the question if editing of the α3 transcript was temporally associated with the suggested developmental shift from excitation to inhibition in the GABA system. The editing frequency was studied by using Sanger and Pyrosequencing, and to monitor the temporal aspects, we studied the messenger RNA expression of the GABAA receptor subunits and chloride pumps, known to be involved in the switch. The results showed that the chick α3 subunit was subjected to RNA editing, and its expression was restricted to cells in the inner nuclear and ganglion cell layer in the retina. The extent of editing increased during development (after embryonic days 8–9) concomitantly with an increase of expression of the chloride pump KCC2. Expression of several GABAA receptor subunits known to mediate synaptic GABA actions was upregulated at this time. We conclude that editing of the chick GABAA subunit α3 transcript in chick retina gives rise to an amino acid change that may be of importance in the switch from excitatory to inhibitory receptors.

Mammalian retinae express multiple connexins that mediate the metabolic and electrical coupling of various cell types. In retinal neurons, only connexin36, connexin45, connexin50, and connexin57 have been described so far. Here, we present an analysis of a novel retinal connexin, connexin30.2 (Cx30.2), and its regulation in the mouse retina. To analyze the expression of Cx30.2, we used a transgenic mouse line in which the coding region of Cx30.2 was replaced by lacZ reporter DNA. We detected the lacZ signal in the nuclei of neurons located in the inner nuclear layer and the ganglion cell layer (GCL). In this study, we focused on the GCL and characterized the morphology of the Cx30.2-expressing cells. Using immunocytochemistry and intracellular dye injections, we found six different types of Cx30.2-expressing ganglion cells: one type of ON-OFF, three types of OFF, and two types of ON ganglion cells; among the latter was the RGA1 type. We show that RGA1 cells were heterologously coupled to numerous displaced amacrine cells. Our results suggest that these gap junction channels may be heterotypic, involving Cx30.2 and a connexin yet unidentified in the mouse retina. Gap junction coupling can be modulated by protein kinases, a process that plays a major role in retinal adaptation. Therefore, we studied the protein kinase–induced modulation of coupling between RGA1 and displaced amacrine cells. Our data provide evidence that coupling of RGA1 cells to displaced amacrine cells is mediated by Cx30.2 and that the extent of this coupling is modulated by protein kinase C.

The rate-limiting step in the recovery of the photoreceptor light response is the hydrolysis of GTP by transducin, a reaction that is accelerated by the RGS9–Gβ5 complex, and its membrane anchor, R9AP. Similar complexes, including RGS7, RGS11, and Gβ5, are found in retinal ON-bipolar cell dendrites. Here, we present evidence that R9AP is also expressed in the dendritic tips of ON-bipolar cells. Immunofluorescent staining for R9AP revealed a punctate pattern of labeling in the outer plexiform layer, where it colocalized with mGluR6. In photoreceptors, R9AP is required for proteolytic stability of the entire regulator of G protein signaling complex, and we found that genetic deletion of R9AP also results in a marked reduction in the levels of RGS11 and Gβ5 in the bipolar cell dendrites; the level of RGS7 was unaffected, suggesting the presence of another interaction partner to stabilize RGS7. To determine the effect of R9AP deletion on the response kinetics of ON-bipolar cells, we compared the electroretinogram (ERG) between wild-type and R9AP-deficient mice. The ERG b-wave, reflecting ON-bipolar cell activity, was delayed and larger in the R9AP-deficient mice. Our data indicate that R9AP is required for stable expression of RGS11–Gβ5 in ON-bipolar cell dendrites. Furthermore, they suggest that the RGS11–Gβ5–R9AP complex accelerates the initial ON-bipolar cell response to light.

Starburst amacrine cells (SACs), the only acetylcholine (ACh)-releasing amacrine cells (ACs) in adult rabbit retina, contain GABA and are key elements in the retina’s directionally selective (DS) mechanism. Unlike many other GABAergic ACs, they use glutamic acid decarboxlyase (GAD)67, not GAD65, to synthesize GABA. Using immunocytochemistry, we demonstrate the apoptosis at birth (P0) of transitory putative ACs that exhibit immunoreactivity (IR) for the ACh-synthetic enzyme choline acetyltransferase (ChAT), GAD67, and the GABA transporter, GAT1. Only a few intact, displaced ChAT-immunoreactive SAC bodies are detected at P0. At P2, ChAT-IR is detected in the two narrowly stratified substrata of starburst dendrites in the inner plexiform layer (IPL). Quantitative analysis reveals that in the first postnatal week, only a small fraction of SACs cells express ChAT- and GABA-IR. Not until the end of the second week are they expressed in all SACs. At P0, a three-tiered stratification of GABA-IR is present in the IPL, entirely different from the adult pattern of seven substrata, emerging at P3–P4, and optimally visualized at P13. At P0, GAD65 is detectable in normally placed AC bodies. At P1, GAD65-IR appears in dendrites of nonstarburst GABAergic ACs, and by P5 is robust in the adult pattern of four substrata in the IPL. GAD65-IR never co-localizes with ChAT-IR. In a temporal comparison of our data with physiological, pharmacological, and ultrastructural studies, we suggest that transitory ChAT-immunoreactive cells share with SACs production of stage II (nicotinic) waves of previsual synchronous activity in ganglion cells (GCs). Further, we conclude that (1) GAD65-immunoreactive, non-SAC GABAergic ACs are the most likely candidates responsible for the suppression of stage III (muscarinic/AMPA-kainate) waves and (2) DS responses first appear in DS GCs, when about 50% of SACs express ChAT- and GABA-IR, and in 100% of DS GCs, when expression occurs in all SACs.

The bird visual system includes a substantial projection, of unknown function, from a midbrain nucleus to the contralateral retina. Every centrifugal, or efferent, neuron originating in the midbrain nucleus makes synaptic contact with the soma of a single unique amacrine cell, the target cell (TC). By labeling efferent neurons in the midbrain, we have been able to identify their terminals in retinal slices and make patch-clamp recordings from TCs. TCs generate Na+-based action potentials (APs) triggered by spontaneous EPSPs originating from multiple classes of presynaptic neurons. Exogenously applied glutamate elicited inward currents having the mixed pharmacology of NMDA, kainate, and inward rectifying AMPA receptors. Exogenously applied GABA elicited currents entirely suppressed by GABAzine and therefore mediated by GABAA receptors. Immunohistochemistry showed the vesicular glutamate transporter, vGluT2, to be present in the characteristic synaptic boutons of efferent terminals, whereas the GABA synthetic enzyme, GAD, was present in much smaller processes of intrinsic retinal neurons. Extracellular recording showed that exogenously applied GABA was directly excitatory to TCs and, consistent with this, NKCC, the Cl− transporter often associated with excitatory GABAergic synapses, was identified in TCs by antibody staining. The presence of excitatory retinal input to TCs implies that TCs are not merely slaves to their midbrain input; instead, their output reflects local retinal activity and descending input from the midbrain.

Transient uniformity detectors (UDs) are a unique type of retinal ganglion cell (RGC) whose maintained firing is transiently suppressed by all types of visual stimuli. In this study, we have characterized the dendritic morphology and tracer-coupling pattern of UDs that were labeled by loose-seal electroporation of Neurobiotin following functional identification in the isolated rabbit retina. The UDs have a bistratified dendritic tree, branching near the margins of the inner plexiform layer in stratum 1 (part of the OFF sublamina) and stratum 4/5 (part of the ON sublamina). Characteristically, many of the distal dendrites in the OFF arbor do not terminate there but dive recurrently back to the ON arbor. As a consequence, the ON dendritic arbor is usually twice as large as the OFF dendritic arbor in area. The UDs sometimes show homologous tracer coupling to neighboring RGCs with the same morphology, and from this material, we estimate that the UDs have a threefold dendritic field overlap and a maximum density of ~100 cells/mm2 on the peak visual streak, accounting for ~2% of RGCs in rabbit retina. The UDs also show strong heterologous tracer coupling to a novel type of amacrine cell that costratifies with the ON arbor of the UD. Consistent with their unistratified medium-field morphology, these St4/5 amacrine cells appear to be GABAergic: their somata are immunopositive for GABA but immunonegative for glycine and glycine transporter 1. We compare the dendritic morphology of the UDs to that of other types of bistratified RGCs described in rabbit retina and note that the stratification levels and distinctive recurrent dendrites closely resemble those of the “ON bistratified diving” RGCs. This raises the possibility that there are two types of RGCs with distinctive physiological properties that have almost identical bistratified dendritic morphologies.

We present a series of experiments exploring the effect of chromaticity on reaction time (RT) for a variety of stimulus conditions, including chromatic and luminance contrast, luminance, and size. The chromaticity of these stimuli was varied along a series of vectors in color space that included the two chromatic-opponent-cone axes, a red–green (L–M) axis and a blue–yellow [S − (L + M)] axis, and intermediate noncardinal orientations, as well as the luminance axis (L + M). For Weber luminance contrasts above 10–20%, RTs tend to the same asymptote, irrespective of chromatic direction. At lower luminance contrast, the addition of chromatic information shortens the RT. RTs are strongly influenced by stimulus size when the chromatic stimulus is modulated along the [S − (L + M)] pathway and by stimulus size and adaptation luminance for the (L–M) pathway. RTs are independent of stimulus size for stimuli larger than 0.5 deg. Data are modeled with a modified version of Pieron’s formula with an exponent close to 2, in which the stimulus intensity term is replaced by a factor that considers the relative effects of chromatic and achromatic information, as indexed by the RMS (square-root of the cone contrast) value at isoluminance and the Weber luminance contrast, respectively. The parameters of the model reveal how RT is linked to stimulus size, chromatic channels, and adaptation luminance and how they can be interpreted in terms of two chromatic mechanisms. This equation predicts that, for isoluminance, RTs for a stimulus lying on the S-cone pathway are higher than those for a stimulus lying on the L–M-cone pathway, for a given RMS cone contrast. The equation also predicts an asymptotic trend to the RT for an achromatic stimulus when the luminance contrast is sufficiently large.

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.

Retinal ganglion cells (RGCs) are highly sensitive to changes in contrast, which is crucial for the detection of edges in a visual scene. However, in the natural environment, edges do not just vary in contrast, but edges also vary in the degree of blur, which can be caused by distance from the plane of fixation, motion, and shadows. Hence, blur is as much a characteristic of an edge as luminance contrast, yet its effects on the responses of RGCs are largely unexplored.

We examined the responses of rabbit RGCs to sharp edges varying by contrast and also to high-contrast edges varying by blur. The width of the blur profile ranged from 0.73 to 13.05 deg of visual angle. For most RGCs, blurring a high-contrast edge produced the same pattern of reduction of response strength and increase in latency as decreasing the contrast of a sharp edge. In support of this, we found a significant correlation between the amount of blur required to reduce the response by 50% and the size of the receptive fields, suggesting that blur may operate by reducing the range of luminance values within the receptive field. These RGCs cannot individually encode for blur, and blur could only be estimated by comparing the responses of populations of neurons with different receptive field sizes. However, some RGCs showed a different pattern of changes in latency and magnitude with changes in contrast and blur; these neurons could encode blur directly.

We also tested whether the response of a RGC to a blurred edge was linear, that is, whether the response of a neuron to a sharp edge was equal to the response to a blurred edge plus the response to the missing spatial components that were the difference between a sharp and blurred edge. Brisk-sustained cells were more linear; however, brisk-transient cells exhibited both linear and nonlinear behavior.

Nonarteritic anterior ischemic optic neuropathy (NAION) is associated with vascular risk factors and a genetic predisposition for NAION. In this study, we examined the potential association of endothelial nitric oxide synthase (eNOS) G894T polymorphism with NAION. For this, 45 patients (29 men and 16 women) and 193 controls (122 men and 71 women) were enrolled prospectively and genotyped for eNOS genes. Genotypes were determined by polymerase chain reaction and restriction enzyme analysis. The prevalence of eNOS polymorphisms was estimated in NAION patients and controls. Genotype frequencies were estimated with chi-square test, and odds ratios were calculated. We found that eNOS G894T polymorphism is not associated with NAION occurrence as the genotype and allele frequencies were not significantly different between the control and patient groups (TT vs. GG + GT: P = 0.646 and T vs. G: P = 0.86). The precise mechanism of NAION occurrence has not been elucidated yet; since NAION may occur when a compromised watershed microcirculation is combined with insufficient autoregulation of systematic circulation, other alterations in the eNOS gene or polymorphism of genes involved in systematic circulation may be associated with NAION occurrence.

The ability to extract form information from a visual scene, for object recognition or figure–ground segregation, is a fundamental visual system function. Many studies of nonhuman primates have addressed the neural mechanisms involved in global form processing, but few have sought to demonstrate this ability behaviorally. In this study, we probed global visual processing in macaque monkeys (Macaca nemestrina) using classical Kanizsa illusory shapes as an assay of global form perception. We trained three monkeys on a “similarity match-to-sample” form discrimination task, first with complete forms embedded in fields of noncontour-inducing “pacman” elements. We then tested them with classic Kanizsa illusory shapes embedded in fields of randomly oriented elements. Two of the three subjects reached our criterion performance level of 80% correct or better on four of five illusory test conditions, demonstrating clear evidence of Kanizsa illusory form perception; the third subject mastered three of five conditions. Performance limits for illusory form discrimination were obtained by manipulating support ratio and by measuring threshold for discriminating “fat” and “thin” illusory squares. Our results indicate that macaque monkeys are capable of global form processing similarly to humans and that the perceptual mechanisms for “filling-in” contour gaps exist in macaques as they do in humans.

Orientation adaptation and perceptual learning change orientation tuning curves of V1 cells. Adaptation shifts tuning curve peaks away from the adapted orientation, reduces tuning curve slopes near the adapted orientation, and increases the responses on the far flank of tuning curves. Learning an orientation discrimination task increases tuning curve slopes near the trained orientation. These changes have been explained previously in a recurrent model (RM) of orientation selectivity. However, the RM generates only complex cells when they are well tuned, so that there is currently no model of orientation plasticity for simple cells. In addition, some feedforward models, such as the modified feedforward model (MFM), also contain recurrent cortical excitation, and it is unknown whether they can explain plasticity. Here, we compare plasticity in the MFM, which simulates simple cells, and a recent modification of the RM (MRM), which displays a continuum of simple-to-complex characteristics. Both pre- and postsynaptic-based modifications of the recurrent and feedforward connections in the models are investigated. The MRM can account for all the learning- and adaptation-induced plasticity, for both simple and complex cells, while the MFM cannot. The key features from the MRM required for explaining plasticity are broadly tuned feedforward inputs and sharpening by a Mexican hat intracortical interaction profile. The mere presence of recurrent cortical interactions in feedforward models like the MFM is insufficient; such models have more rigid tuning curves. We predict that the plastic properties must be absent for cells whose orientation tuning arises from a feedforward mechanism.

Eye fixation–related potential (EFRP) measures electrical brain activity in response to eye fixations. The aim of the current study was to investigate whether the EFRPs vary during consecutive eye fixations while subjects were performing an object identification task. Eye fixations evoked P1 and N1 components at the occipital and parietal recording sites. The latency of P1 component increased during consecutive fixations. The amplitude of P1 increased and the amplitude of N1 decreased during consecutive fixations. The results indicate that EFRPs are modulated during consecutive fixations, suggesting that the current technique may provide a useful tool to study temporal dynamics of visual perception and processes underlying object identification.