An Alternative Splice Variant of Zebrafish Cx52.6 is Expressed in Retinal Horizontal Cells.

Retinal horizontal cells (HCs) are inhibitory neurons, which modulate the transmission of light-elicited signals from photoreceptors to bipolar cells in the outer retina. HCs of the same physiological type are extensively coupled via gap junctions. In the zebrafish retina, the population of HCs comprises up to four morphologically distinct subtypes. Four different connexins (Cx52.6, Cx52.7, Cx52.9 and Cx55.5) were detected in these cells with overlapping expression patterns. In this study, we show that Cx52.6 is alternatively spliced in the retina, resulting in an additional isoform, designated as Cx53.4, which differs from the originally described Cx52.6 only by the final C-terminal peptide (12 vs. 4 aa). Further protein sequence alignments revealed that Cx53.4 represents the counterpart of alternatively spliced mouse Cx57 and human Cx62. RT-PCR analyses of mRNA expression in different adult zebrafish tissues showed that Cx53.4 is expressed exclusively in the retina. The localization of Cx53.4 protein within the retina was analyzed using a specific antibody. Immunofluorescence analyses demonstrated that the expression of Cx53.4 is restricted to HCs of all four subtypes. Further, immunoelectron microscopy confirmed the presence of Cx53.4 in gap junctions between HC dendrites and between their axon terminals.

Cones perform a non-linear transformation on natural stimuli.

Visual information in natural scenes is distributed over a broad range of intensities and contrasts. This distribution has to be compressed in the retina to match the dynamic range of retinal neurons. In this study we examined how cones perform this compression and investigated which physiological processes contribute to this operation. M- and L-cones of the goldfish were stimulated with a natural time series of intensities (NTSI) and their responses were recorded. The NTSI displays an intensity distribution which is skewed towards the lower intensities and has a long tail into the high intensity region. Cones transform this skewed distribution into a more symmetrical one. The voltage responses of the goldfish cones were compared to those of a linear filter and a non-linear biophysical model of the photoreceptor. The results show that the linear filter under-represents contrasts at low intensities compared to the actual cone whereas the non-linear biophysical model performs well over the whole intensity range used. Quantitative analysis of the two approaches indicates that the non-linear biophysical model can capture 91 +/- 5% of the coherence rate (a biased measure of information rate) of the actual cone, where the linear filter only reaches 48 +/- 8%. These results demonstrate that cone photoreceptors transform an NTSI in a non-linear fashion. The comparison between current clamp and voltage clamp recordings and analysis of the behaviour of the biophysical model indicates that both the calcium feedback loop in the outer segment and the hydrolysis of cGMP are the major components that introduce the specific non-linear response properties found in the goldfish cones.

Nightblindness-associated transient tonic downgaze (NATTD) in infant boys with chin-up head posture.

Eleven infant boys presented with chin-up head posture, tonic downgaze and, on attempted upgaze, large-amplitude upward saccades with deceleration during the slow phase downward. The gaze-evoked upward saccades disappeared at the age of 2 or 3 years. In addition, they had high-frequency, small-amplitude horizontal pendular nystagmus that remained. Among these infant boys were 2 pairs of maternally related half-brothers, 2 cousins, and 2 siblings. Visual acuity ranged from 0.1 to 0.6, ERG-amplitudes (both A- and B-wave) were reduced, and severe myopia was found in 5 cases. Eight boys had CACNA1F mutations, and 1 boy had a NYX mutation, compatible with incomplete or complete congenital stationary nightblindness (iCSNB or cCSNB), respectively. This points to a defective synapse between the rod and the ON-bipolar cell causing the motility disorder: CACNA1F is located on the rod side of this synapse, whereas NYX is located on the side of the ON-bipolar cell. The coexistence of horizontal and vertical nystagmus has been previously described in dark-reared cats.

Pannexin1 in the outer retina of the zebrafish, Danio rerio.

In the retina, chemical and electrical synapses couple neurons into functional networks. New candidates encoding for electrical synapse proteins have recently emerged. In the present study, we determined the localization of the candidate protein pannexin1 (zfPanx1) in the zebrafish retina and studied the functional properties of zfPanx1 exogenously expressed in Neuroblastoma 2a (N2a) cells. zfPanx1 was identified on the surface of horizontal cell dendrites invaginating deeply into the cone pedicle near the glutamate release sites of the cones, providing in vivo evidence for hemichannel formation at that location. This strategic position of zfPanx1 in the photoreceptor synapse could potentially allow modulation of cone output. Using whole cell voltage clamp and excised patch recordings of transfected N2a cells, we demonstrated that zfPanx1 forms voltage-activated hemichannels with a large unitary conductance in vitro. These channels can open at physiological membrane potentials. Functional channels were not formed following mutation of a single amino acid within a conserved protein motif recently shown to be N-glycosylated in rodent Panx1. Together, these findings indicate that zfPanx1 displays properties similar to its mammalian homologues and can potentially play an important role in functions of the outer retina.

The contribution of the outer retina to color constancy: a general model for color constancy synthesized from primate and fish data.

Color constancy is one of the most impressive features of color vision systems. Although the phenomenon has been studied for decades, its underlying neuronal mechanism remains unresolved. Literature indicates an early, possibly retinal mechanism and a late, possibly cortical mechanism. The early mechanism seems to involve chromatic spatial integration and performs the critical calculations for color constancy. The late mechanism seems to make the color manifest. We briefly review the current evidence for each mechanism. We discuss in more detail a model for the early mechanism that is based on direct measurements of goldfish outer retinal processing and induces color constancy and color contrast. In this study we extrapolate this model to primate retina, illustrating that it is highly likely that a similar mechanism is also present in primates. The logical consequence of our experimental work in goldfish and our model is that the wiring of the cone/horizontal cell system sets the reference point for color vision (i.e., it sets the white point for that animal).

The involvement of glutamate-gated channels in negative feedback from horizontal cells to cones.

Photoreceptors are the light sensitive cells in the retina. They project to horizontal cells and bipolar cells via a glutamatergic feed forward pathway. Horizontal cells are strongly electrically coupled and integrate in that way the input from the photoreceptors. Horizontal cells feedback to cones negatively. The combined signal from the photoreceptors and the horizontal cells is sent to the bipolar cells. The feedback pathway from horizontal cells to cones is thought to form the basis for the center/surround organization of bipolar cells. The nature of the feedback pathway is an issue of intense debate. It was thought for a long time that this feedback pathway was GABAergic, because cones have GABA-receptors and horizontal cells release GABA via a GABA-transporter working in the reversed direction. However, recently we showed in goldfish that horizontal cells feed back to cones via an alternative mechanism. In goldfish, negative feedback from horizontal cells to cones shifts the calcium current of the cone to more negative potentials. This feedback pathway is independent of GABA, since feedback cannot be blocked by either saturating concentrations of PTX, the GABA-transporter blocker SKF89976A, or application of GABA. The mechanism of negative feedback from horizontal cells to cones involves hemichannels located at the tips of the invaginating horizontal cells, just opposite to the calcium channels of the cones. Current flowing through these hemichannels changes the extracellular potential deep in the synaptic cleft and in that way modulates the calcium current of the cones. Such a modulation of the extracellular potential is called ephaptic. If negative feedback from horizontal cells to cones is indeed ephaptic, other channels present in the synapse should also be able to act as a current source, i.e., should also be able to change the output of the cone. We showed that glutamate-gated channels present at the tips of the horizontal cell dendrites can also mediate feedback responses. Surprisingly, although the glutamate-gated conductance of the horizontal cells is eight times the hemichannel conductance, glutamate-gated channels are not the major current source in negative feedback from horizontal cells to cones. In this chapter we present evidence that this is due to the more focal localization of the hemichannels, compared to a diffuse and extrasynaptic localization of the glutamate-gated channels.

Cobalt ions inhibit negative feedback in the outer retina by blocking hemichannels on horizontal cells.

In goldfish, negative feedback from horizontal cells to cones shifts the activation function of the Ca2+ current of the cones to more negative potentials. This shift increases the amount of Ca2+ flowing into the cones, resulting in an increase in glutamate release. The increased glutamate release forms the basis of the feedback-mediated responses in second-order neurons, such as the surround-induced responses of bipolar cells and the spectral coding of horizontal cells. Low concentrations of Co2+ block these feedback-mediated responses in turtle retina. The mechanism by which this is accomplished is unknown. We studied the effects of Co2+ on the cone/horizontal network of goldfish retina and found that Co2+ greatly reduced the feedback-mediated responses in both cones and horizontal cells in a GABA-independent way. The reduction of the feedback-mediated responses is accompanied by a small shift of the Ca2+ current of the cones to positive potentials. We have previously shown that hemichannels on the tips of the horizontal cell dendrites are involved in the modulation of the Ca2+ current in cones. Both the absence of this Co2+-induced shift of the Ca2+ current in the absence of a hemichannel conductance and the sensitivity of Cx26 hemichannels to low concentrations of Co2+ are consistent with a role for hemichannels in negative feedback from horizontal cells to cones.

The dynamic characteristics of the feedback signal from horizontal cells to cones in the goldfish retina.

1. The dynamic properties of the microcircuitry formed by cones and horizontal cells in the isolated goldfish retina were studied. Cones project to horizontal cells and horizontal cells feed back to cones via a relatively slow negative feedback pathway. 2. The time constant of the feedback signal in cones and of the effect this feedback signal had on the responses of second-order neurons was determined using whole-cell patch clamp and intracellular recording techniques. 3. It was found that the feedback signal in cones had a time constant of around 80 ms, whereas the time constant of the effect this feedback signal had on the second-order neurons ranged from 36 to 116 ms. This range of time constants can be accounted for by the non-linearity of the Ca(2+) current in the cones. In depolarized cones, the feedback-mediated response in second-order neurons had a similar time constant to that of the direct light response of the cone, whereas in hyperpolarized cones, the time constant of the feedback-mediated response in second-order neurons was considerably larger. 4. Further, it was shown that there was no delay in the feedback pathway. This is in contrast to what has been deduced from the response properties of second-order neurons. In one type of horizontal cell, the responses to red light were delayed relative to the responses to green light. This delay in the second-order neurons can be accounted for by the interaction of the direct light response of the medium-wavelength-sensitive cones (M-cones) with the feedback response of the M-cones received from the horizontal cells.

Hemichannel-mediated inhibition in the outer retina.

An essential feature of the first synapse in the retina is a negative feedback pathway from horizontal cells to cones. Here we show that at this synapse, connexin26 forms hemichannels on horizontal cell dendrites near the glutamate release site of the cones. Blocking these hemichannels hyperpolarizes horizontal cells, modulates the Ca2+ channels of the cones, and abolishes all feedback-mediated responses. We propose a feedback mechanism in which the activity of the Ca2+ channels and the subsequent glutamate release of the cones are modulated by a current through these hemichannels. Because the current through the hemichannels depends on the polarization of the horizontal cells, their activity modulates the output of the cones.

The value of electrophysiology results in patients with epilepsy and vigabatrin associated visual field loss.

PURPOSE: To determine the value of electrophysiological findings in patients with temporal lobe epilepsy and to relate these findings to the amount of concentric contraction of the visual field and the use of vigabatrin. METHODS: Electro-retinograms and electro-oculograms were done on 30 patients, operated for temporal lobe epilepsy. The patients were divided into three groups: (A) concentric contraction of the visual field associated with a history of vigabatrin medication (15 patients), (B) normal visual field with vigabatrin use (11 patients) and (C) normal visual field without vigabatrin medication (4 patients). RESULTS: Electrophysiological abnormalities were found in 50% of the patients in group A. The Arden ratio of the EOG was lowered in 57%. Abnormalities in the ERG were found: b-wave implicit time photopic F was prolonged (50%), b-wave amplitudes scotopic B (53%), C (73%) and G (50%) and photopic H (50%) were diminished. The amount of visual field loss and the total dose of vigabatrin used, showed only slight correlation with the ERG and EOG. The use of vigabatrin during the ERG and EOG recording in group A, gave a higher b-wave amplitude scotopic G in 64% of cases. The a-wave implicit times scotopic G (73%) and photopic G (59%) and H (73%) were shortened in group B. CONCLUSION: EOG was abnormal in 57% in group A. ERG abnormalities could only be found in 50% of group A, mainly in the inner retina. Since also the total dose of vigabatrin and the amount of visual field loss did not really show a correlation with the electrophysiological findings and results of literature are not unanimous, electrophysiology does not appear at present to be a good method to detect patients with, or at risk of, vigabatrin associated visual field loss. Regularly performed visual field examination remains the cornerstone in screening.

The open- and closed-loop gain-characteristics of the cone/horizontal cell synapse in goldfish retina.

Under constant light-adapted conditions, vision seems to be rather linear. However, the processes underlying the synaptic transmission between cones and second-order neurons (bipolar cells and horizontal cells) are highly nonlinear. In this paper, the gain-characteristics of the transmission from cones to horizontal cells and from horizontal cells to cones are determined with and without negative feedback from horizontal cells to cones. It is shown that 1) the gain-characteristic from cones to horizontal cells is strongly nonlinear without feedback from horizontal cells, 2) the gain-characteristic between cones and horizontal cells becomes linear when feedback is active, and 3) horizontal cells feed back to cones via a linear mechanism. In a quantitative analysis, it will be shown that negative feedback linearizes the synaptic transmission between cones and horizontal cells. The physiological consequences are discussed.

Immunocytochemical localization of the glutamate transporter GLT-1 in goldfish (Carassius auratus) retina.

Glutamate is the major excitatory neurotransmitter in the retina of vertebrates. Electrophysiological experiments in goldfish and salamander have shown that neuronal glutamate transporters play an important role in the clearance of glutamate from cone synaptic clefts. In this study, the localization of the glutamate transporter GLT-1 has been investigated immunocytochemically at the light and electron microscopical levels in the goldfish retina using a GLT-1-specific antibody. GLT immunoreactivity (IR) was observed at the light microscopical level in Müller cells, bipolar cells, the outer plexiform layer (OPL), and the inner plexiform layer (IPL). At the electron microscopical level, membrane-bound and cytoplasmic GLT-IR in the OPL was located in finger-like protrusions of the cone terminal located near the invaginating postsynaptic processes of bipolar and horizontal cells. GLT-IR was not observed in the vicinity of synaptic ribbons. This location of GLT-1 allows modulation of the glutamate concentration in the synaptic cleft, thereby shaping the dynamics of synaptic transmission between cones and second-order neurons. In the inner IPL, GLT-IR was observed in the cytoplasm and was membrane bound in mixed rod/cone bipolar cell terminals and cone bipolar cell terminals. The membrane-bound GLT-1 was generally observed at some distance from the synaptic ribbon. The morphology of the bipolar cell terminal together with the localization of GLT-1 suggests that at least these glutamate transporters are not primarily involved in rapid uptake of glutamate release by the bipolar cells. The GLT-IR in the cytoplasm of Müller cells was located throughout the entire goldfish retina from the outer limiting membrane to the inner limiting membrane. The location of GLT-1 in Müller cells is consistent with the role of Müller cells in converting glutamate to glutamine.

Expression and localization of ionotropic glutamate receptor subunits in the goldfish retina--an in situ hybridization and immunocytochemical study.

The expression and distribution of AMPA, kainate and NMDA glutamate receptor subunits was studied in the goldfish retina. For the immunocytochemical localization of the AMPA receptor antisera against GluR2, GluR2/3 and GluR4 were used, and for in situ hybridization rat specific probes for GluR1 and GluR2 and goldfish specific probes for GluR3 and GluR4 were used. The localization of the low affinity kainate receptor and NMDA receptor was studied using antisera against GluR5-7 and NR1. All AMPA receptor subtypes were demonstrated to be present in the goldfish retina both by immunocytochemistry and in situ hybridization. In situ hybridization revealed expression of all AMPA receptors subunit at the inner border of the INL. Only GluR3 was also strongly expressed in the outer border of the INL. Some of the ganglion cells displayed a strong signal for GluR1, GluR3 and GluR4. GluR1-immunoreactivity was present in subsets of bipolar, amacrine, and ganglion cells. GluR2 and GluR2/3-immunoreactivity was mainly localized in the outer plexiform layer. GluR2 and GluR2/3-immunoreactivity are associated with the photoreceptor synaptic terminals. GluR4-immunoreactivity is present on Müller cells in the inner retina and on dendrites of bipolar cells in the OPL, whereas GluR5-7-immunoreactivity was prominently present on horizontal cell axon terminals. Finally, NR1-immunoreactivity was confined to amacrine cells, the inner plexiform layer and ganglion cells. This study shows that there is a strong heterogeneity of glutamate receptor subunit expression in the various layers of the retina. Of the AMPA receptor subunits GluR3 seems to be expressed the most widely in all layers with strong glutamatergic synaptic interactions whereas all the other subunits seem to have a more restricted expressed pattern.

The nature of surround-induced depolarizing responses in goldfish cones.

Cones in the vertebrate retina project to horizontal and bipolar cells and the horizontal cells feedback negatively to cones. This organization forms the basis for the center/surround organization of the bipolar cells, a fundamental step in the visual signal processing. Although the surround responses of bipolar cells have been recorded on many occasions, surprisingly, the underlying surround-induced responses in cones are not easily detected. In this paper, the nature of the surround-induced responses in cones is studied. Horizontal cells feed back to cones by shifting the activation function of the calcium current in cones to more negative potentials. This shift increases the calcium influx, which increases the neurotransmitter release of the cone. In this paper, we will show that under certain conditions, in addition to this increase of neurotransmitter release, a calcium-dependent chloride current will be activated, which polarizes the cone membrane potential. The question is, whether the modulation of the calcium current or the polarization of the cone membrane potential is the major determinant for feedback-mediated responses in second-order neurons. Depolarizing light responses of biphasic horizontal cells are generated by feedback from monophasic horizontal cells to cones. It was found that niflumic acid blocks the feedback-induced depolarizing responses in cones, while the shift of the calcium current activation function and the depolarizing biphasic horizontal cell responses remain intact. This shows that horizontal cells can feed back to cones, without inducing major changes in the cone membrane potential. This makes the feedback synapse from horizontal cells to cones a unique synapse. Polarization of the presynaptic (horizontal) cell leads to calcium influx in the postsynaptic cell (cone), but due to the combined activity of the calcium current and the calcium-dependent chloride current, the membrane potential of the postsynaptic cell will be hardly modulated, whereas the output of the postsynaptic cell will be strongly modulated. Since no polarization of the postsynaptic cell is needed for these feedback-mediated responses, this mechanism of synaptic transmission can modulate the neurotransmitter release in single synaptic terminals without affecting the membrane potential of the entire cell.

Effect of the tuberculostaticum ethambutol and stimulus intensity on chromatic discrimination in man.

In goldfish it has been shown that ethambutol shifts the threshold for wavelength discrimination without affecting the absolute sensitivity of the cones. In this study we demonstrate that a similar colour vision disturbance occurs in tuberculosis patients treated with ethambutol. After 2 months of ethambutol treatment, chromatic discrimination was measured with a computerized forced two choice (CD) test with isoluminant coloured stimuli and with three other colour vision tests: the Ishihara, the Oscar and the Lanthony Desaturated 15 Hue tests. The scores of the patient group (n = 19) on these four colour vision tests were compared with the scores of a group of control subjects (n = 33) and a group of congenital red/green colour-blind subjects (n = 5). A reduction of the stimulus intensity of 1 log unit caused a significant reduction in red/green chromatic discrimination, measured with the CD test in both, control subjects and patients. This intensity dependent reduction was significantly greater for patients than for controls. In this respect, man and goldfish behave similarly. Furthermore, the CD test showed the same ethambutol-induced reduction in chromatic discrimination at low intensity for the blue/green part of the spectrum. This has not been measured in goldfish. The origin of this ethambutol-induced colour vision disturbance must be at a post-photoreceptor site, because the Ishihara and Oscar tests, both designed to screen for photoreceptor-based, or primary red/green colour vision disturbances, did not discriminate between patients and control subjects. Thus, as in goldfish, we find that in patients ethambutol shifts the threshold for chromatic discrimination without changing the absolute sensitivity.

Intrinsic cone adaptation modulates feedback efficiency from horizontal cells to cones.

Processing of visual stimuli by the retina changes strongly during light/dark adaptation. These changes are due to both local photoreceptor-based processes and to changes in the retinal network. The feedback pathway from horizontal cells to cones is known to be one of the pathways that is modulated strongly during adaptation. Although this phenomenon is well described, the mechanism for this change is poorly characterized. The aim of this paper is to describe the mechanism for the increase in efficiency of the feedback synapse from horizontal cells to cones. We show that a train of flashes can increase the feedback response from the horizontal cells, as measured in the cones, up to threefold. This process has a time constant of approximately 3 s and can be attributed to processes intrinsic to the cones. It does not require dopamine, is not the result of changes in the kinetics of the cone light response and is not due to changes in horizontal cells themselves. During a flash train, cones adapt to the mean light intensity, resulting in a slight (4 mV) depolarization of the cones. The time constant of this depolarization is approximately 3 s. We will show that at this depolarized membrane potential, a light-induced change of the cone membrane potential induces a larger change in the calcium current than in the unadapted condition. Furthermore, we will show that negative feedback from horizontal cells to cones can modulate the calcium current more efficiently at this depolarized cone membrane potential. The change in horizontal cell response properties during the train of flashes can be fully attributed to these changes in the synaptic efficiency. Since feedback has major consequences for the dynamic, spatial, and spectral processing, the described mechanism might be very important to optimize the retina for ambient light conditions.

The feedback pathway from horizontal cells to cones. A mini review with a look ahead.

The feedback pathway from HCs to cones forms the basis of the surround responses of the bipolar cells and is essential for the spectral opponency of horizontal cells. The nature of this feedback pathway is an issue of debate. Three hypothesis are presented in literature: (1) a GABAA-ergic feedback pathway; (2) a GABA-independent feedback pathway that modulates the Ca-current in cones; and (3) an electrical feedback pathway. In this review the evidence for the various pathways will be discussed. The conclusion is that the available evidence favors the hypothesis that feedback modulates the Ca-current in the cones in a GABA independent way. An alternative role of GABA in the outer plexiform layer is discussed and finally the functional consequences of the negative feedback pathway from horizontal cells to cones are presented.

Spectral sensitivity of the feedback signal from horizontal cells to cones in goldfish retina.

The spectral sensitivity of cones in isolated goldfish retina was determined with whole-cell recording techniques. Three spectral classes of cones were found with maximal sensitivities around 620 nm, 540 nm, and 460 nm. UV-cones were not found because our stimulator did not allow effective stimulation in the UV range. The spectral sensitivity of the cones closely matched the cone photopigment absorption spectra at the long wavelength side of the spectrum, but deviated significantly at shorter wavelengths. Surround stimulation induced an inward current in cones due to feedback from horizontal cells. The spectral sensitivity of this feedback signal was determined in all three cone classes and found to be broader than the spectral sensitivity of the cones recorded from, and to be spectrally nonopponent. These data are consistent with a connectivity scheme between cones and horizontal cells in which the three horizontal cell systems feed back to all cone systems and in which all horizontal cell systems receive input from more than one cone system.

The cone/horizontal cell network: a possible site for color constancy.

Color vision is spectrally opponent, suggesting that spectrally opponent neurons, such as the horizontal cells in fish and turtle retinae, play a prominent role in color discrimination. In the accompanying paper (Kraaij et al., 1998), it was shown that the output signal of the horizontal cell system to the cones is not at all spectrally opponent. Therefore, a role for the spectrally opponent horizontal cells in color discrimination seems unlikely. In this paper, we propose that the horizontal cells play a prominent role in color constancy and simultaneous color contrast instead of in color discrimination. We have formulated a model of the cone/horizontal cell network based on measurements of the action spectra of the cones and of the feedback signal of the horizontal cell system to the various cone types. The key feature of the model is (1) that feedback is spectrally and spatially very broad and (2) that the gain of the cone synapse strongly depends on the feedback strength. This makes the synaptic gain of the cones strongly dependent on the spectral composition of the surround. Our model, which incorporates many physiological details of the outer retina, displays a behavior that can be interpreted as color constancy and simultaneous color contrast. We propose that the horizontal cell network modulates the cone synaptic gains such that the ratios of the cone outputs become almost invariant with the spectral composition of the global illumination. Therefore, color constancy appears to be coded in the retina.

GABA sensitivity of spectrally classified horizontal cells in goldfish retina.

We studied the GABA sensitivity of horizontal cells in the isolated goldfish retina. After the glutamatergic input to the horizontal cells was blocked with DNQX, GABA depolarized the monophasic and biphasic horizontal cells. The pharmacology of these GABA-induced depolarizations was tested with the GABA receptor antagonists bicuculline-methiodide and picrotoxin, the GABA transporter agonist nipecotic acid, and the GABA transporter antagonist SKF 89976-A. The GABA-induced responses of monophasic horizontal cells consisted of two components; one with characteristics of GABA-gated chloride channels, and one with characteristics of GABA transporters. In biphasic horizontal cells, we only found evidence for GABA-gated chloride channels. The results show that monophasic horizontal cells in goldfish contain the two components of a positive feedback loop (GABA transporters and GABA-gated chloride channels), as described in salamander. Furthermore, our results indicate that the monophasic horizontal cells may project directly to the biphasic horizontal cells, via an excitatory GABAergic pathway. We propose that the function of these GABAergic systems in horizontal cells is to abolish cone dominance in bipolar cells surround in the dark-adapted retina.