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A Borst

Publications and source records attributed to A Borst.

At least 37 records · Page 2Linked to original sources

Voltage signals of individual Purkinje cell dendrites in rat cerebellar slices.

For investigating neuronal information processing at the cellular level, a technique which visualizes the voltage distribution within single neurons in situ would be extremely useful. Voltage-sensitive dyes are, in principle, capable of reporting membrane potential [Cohen, L.B. and Salzberg, B.M., Rev. Physiol. Biochem. Pharmacol., 83 (1978) 35-88; Grinvald, A., Lieke, E.E., Frostig, R.D. and Hildesheim, R., J. Neurosci., 14 (1994) 2545-2568; Kleinfeld, D., Delaney, K.R., Fee, M.S., Flores, J.A., Tank, D.W. and Gelperin, A., J. Neurophysiol., 72 (1994) 1402-1419]. However, their application to single cells internally is technically difficult [Antic, S. and Zecevic, D., J. Neurosci., 15 (1995) 1392-1405; Grinvald, A., Salzberg, B.M., Lev-Ram, V. and Hildesheim, R., Biophys. J., 51 (1987) 643-651; Kogan, A., Ross, W.N., Zecevic, D. and Lasser-Ross, N., Brain Res., 700 (1995) 235-239; Zecevic, D., Nature, 381 (1996) 322-325]. An alternative strategy consists in applying the dye from the outside to all cells in the tissue, while manipulating a single cell by current injection [Krauthamer, V. and Ross, W.N., J. Neurosci., 4 (1984) 673-682; Ross, W.N. and Krauthamer, V., J. Neurosci., 4 (1984) 659-672]. Here, we modify this technique to further enhance spatial at the cost of temporal resolution [Borst, A., Z. Naturforsch., 50 (1995) 435-438]. Applied to rat cerebellar slices we demonstrate that the potential spread in individual Purkinje cells can be imaged up to even fine dendritic branches. The acquired optical signals suggest that steadily hyperpolarized Purkinje cells are electrically compact. When permanently depolarized, the somatic input resistance is significantly diminished, yet the spatial voltage drop along the dendrites remains unchanged. As demonstrated by compartmental modeling, this hints to a concentration of outward rectifying currents at the soma of the cells.

Animals↗

Dendritic computation of direction selectivity and gain control in visual interneurons.

The extraction of motion information from time varying retinal images is a fundamental task of visual systems. Accordingly, neurons that selectively respond to visual motion are found in almost all species investigated so far. Despite its general importance, the cellular mechanisms underlying direction selectivity are not yet understood in most systems. Blocking inhibitory input to fly visual interneurons by picrotoxinin (PTX), we demonstrate that their direction selectivity arises largely from interactions between postsynaptic signals elicited by excitatory and inhibitory input elements, which are themselves only weakly tuned to opposite directions of motion. Their joint activation by preferred as well as null direction motion leads to a mixed reversal potential at which the postsynaptic response settles for large field stimuli. Assuming the activation ratio of these opponent inputs to be a function of pattern velocity can explain how the postsynaptic membrane potential saturates with increasing pattern size at different levels for different pattern velocities ("gain control"). Accordingly, we find that after blocking the inhibitory input by PTX, gain control is abolished.

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gamma-Aminobutyric acid receptor distribution in the mushroom bodies of a fly (Calliphora erythrocephala): a functional subdivision of Kenyon cells?

Antibodies against the Drosophila gamma-aminobutyric acid (GABA) receptor subunit RDL were used to investigate the significance of inhibitory inputs to the mushroom bodies in the blowfly (Calliphora erythrocephala) brain. The pedunculus and the lobes of the mushroom body, which mainly consist of Kenyon cell fibers, revealed strong immunoreactivity against RDL. Pedunculi, alpha- and beta-lobe show characteristic unstained core structures with concentric labeling along the neuropile axis. The gamma-lobes in contrast exhibit a compartmentalized RDL-immunoreactive pattern. These data suggest an important role of GABAergic inhibition in the pedunculus and the lobes of insect mushroom bodies. It is most likely that the RDL-immunoreactivity in the mushroom bodies is closely related to Kenyon cell fibers suggesting that Kenyon cells are an inhomogeneous class of neurons, only part of which receive inhibitory GABAergic input from extrinsic elements. GABAergic inhibition, therefore, may play a substantial role in the process of learning and memory formation in the insect mushroom bodies.

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Encoding of visual motion information and reliability in spiking and graded potential neurons.

We investigated the information about stimulus velocity inherent in the membrane signals of two types of directionally selective, motion-sensitive interneurons in the fly visual system. One of the cells, the H1-cell, is a spiking neuron, whereas the other, the HS-cell, encodes sensory information mainly by a graded shift of its membrane potential. Using a pseudo-random velocity waveform by which a visual grating is moving along the horizontal axis of the eye, both cell types follow the stimulus velocity at higher precision than in response to a step-like velocity function. To measure how much information about the stimulus velocity is preserved in the cellular responses, we calculated the coherence between the stimulus and the neural signals as a function of stimulus frequency. At frequencies up to approximately 10 Hz motion information is well contained in the electrical signals of HS- and H1-cells: For HS-cells the coherence value amounts to approximately 70%, and for H1-cells this value is approximately 60%. Comparing these values with the coherence expected from a linear encoding reveals that the fidelity of the original stimulus is deteriorated in the neural signal partly by neural noise and partly by the nonlinearity inherent in the process of visual motion detection. The degree to which this nonlinearity contributes to the decrease in coherence depends on the maximum velocity used in the experiments; the smaller the stimulus amplitude, the higher the coherence and, thus, the smaller the nonlinearity in encoding of stimulus motion. All these results are in agreement with model simulations in which visual motion is processed by an array of local motion detectors, the spatially integrated output of which is considered the equivalent of the neural signals of HS- and H1-cells.

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Synapse distribution on VCH, an inhibitory, motion-sensitive interneuron in the fly visual system.

In this study, the distribution of synapses in the ventral centrifugal horizontal (VCH)-a nonspiking, inhibitory, motion-sensitive interneuron in the third visual ganglion (lobula plate) of the blowfly Calliphora erythrocephala-was examined by electron microscopy and electrophysiology. The frequency histograms of excitatory and inhibitory postsynaptic potential amplitudes recorded from the VCH during contralateral stimulus presentation suggest the existence of three neurons, two excitatory and one inhibitory, mediating contralateral input. To localize input and output regions of the VCH, we investigated the ultrastructure of its two arborisation areas after intracellular iontophoretic injection of horseradish peroxidase. The VCH has input synapses in its arborisation in the protocerebrum and in its arborisation in the lobula plate. Output synapses were found exclusively in the lobula plate. Thus, the large dendritic arbor of the VCH in the lobula plate serves simultaneously as an input and an output region. There, we found input and output synapses in close vicinity (0.5-1.5 microm) to each other. Taking into account that the VCH receives retinotopicly arranged input from the ipsilateral eye in the lobula plate, the close location of input and output synapses in the VCH suggests that the spatial organization of its retinotopic synaptic input is more or less conserved in its inhibitory output pattern. The VCH has been proposed to inhibit the figure detection 1 (FD1), another neuron of the lobula plate, that responds preferentially to small moving objects. These results suggest that the FD1 may receive inhibitory inputs from the VCH in the lobula plate, where the dendritic arbors of both neurons overlap.

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Comparison of the internal transcribed spacer, ITS 1, from Toxoplasma gondii isolates and Neospora caninum.

The internal transcribed spacer (ITS1) region and the 5' part of the 5.8S ribosomal RNA gene of the ribosomal DNA repeat from 20 Toxoplasma gondil isolates was sequenced and found to be identical in all isolates, independent of host origin or virulence to mice. The ITS1 region from the closely related coccidian parasite Neospora caninum differed in 22% of its nucleotides. Hence, the ITS1 region provides a good marker for the distinction of T. gondii and N. caninum but is not useful for epidemiology studies of T. gondii.

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The intrinsic electrophysiological characteristics of fly lobula plate tangential cells: II. Active membrane properties.

The voltage-gated currents in the fly lobula plate tangential cells were examined using the switched electrode voltage clamp technique. In CH cells, two currents were identified (Figs. 1, 2): a slow calcium inward current and a delayed rectifying, noninactivating potassium outward current. HS and VS cells appear to possess similar currents to CH cells, but in addition, exhibit a fast-activating sodium inward current and a sodium-activated potassium outward current (Figs. 3, 4). While the delayed rectifying potassium current in all three cell classes is responsible for the observed outward rectification described previously (Borst and Haag, 1996), the sodium inward current produced the fast and irregular spikelike depolarizations found in HS and VS cells but not in CH cells: When the sodium current is blocked by either. TTX or intracellular QX314, no more action potentials can be elicited in HS cells under current-clamp conditions (Fig. 5). As is demonstrated in HS cells, space clamp conditions are sufficient to suppress synaptically induced action potentials (Fig. 6). The currents described above were incorporated with the appropriate characteristics into compartmental models of the cells (Fig. 7, 8). The anatomical and electrically passive membrane parameters of these cells were determined in a preceding paper (Borst and Haag, 1996). After fitting the current parameters to the voltage-clamp data (Fig. 9), the model cells qualitatively mimicked the fly tangential cells under current clamp conditions in response to current injection (Fig. 10). The simulations demonstrated that the electrical compactness seen in the HS and VS cells, either in passive models or in active models during continuous hyperpolarization, decreased significantly in the active models during continuous depolarization (Fig. 11). Active HS models reproduce the frequency-dependent amplification of current injected into their axon (Fig. 12).

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The intrinsic electrophysiological characteristics of fly lobula plate tangential cells: I. Passive membrane properties.

The passive membrane properties of the tangential cells in the fly lobula plate (CH, HS, and VS cells, Fig. 1) were determined by combining compartmental modeling and current injection experiments. As a prerequisite, we built a digital base of the cells by 3D-reconstructing individual tangential cells from cobalt-stained material including both CH cells (VCH and DCH cells), all three HS cells (HSN, HSE, and HSS cells) and most members of the VS cell family (Figs. 2, 3). In a first series of experiments, hyperpolarizing and depolarizing currents were injected to determine steady-state I-V curves (Fig. 4). At potentials more negative than resting, a linear relationship holds, whereas at potentials more positive than resting, an outward rectification is observed. Therefore, in all subsequent experiments, when a sinusoidal current of variable frequency was injected, a negative DC current was superimposed to keep the neurons in a hyperpolarized state. The resulting amplitude and phase spectra revealed an average steady-state input resistance of 4 to 5 M omega and a cut-off frequency between 40 and 80 Hz (Fig. 5). To determine the passive membrane parameters Rm (specific membrane resistance), Ri (specific internal resistivity), and Cm (specific membrane capacitance), the experiments were repeated in computer simulations on compartmental models of the cells (Fig. 6). Good fits between experimental and simulation data were obtained for the following values: Rm = 2.5 k omega cm2, Ri = 60 omega cm, and Cm = 1.5 microF/cm2 for CH cells; Rm = 2.0 k omega cm2, Ri = 40 omega cm, and Cm = 0.9 microF/cm2 for HS cells; Rm = 2.0 k omega cm2, Ri = 40 omega cm, and Cm = 0.8 microF/cm2 for VS cells. An error analysis of the fitting procedure revealed an area of confidence in the Rm-Ri plane within which the Rm-Ri value pairs are still compatible with the experimental data given the statistical fluctuations inherent in the experiments (Figs. 7, 8). We also investigated whether there exist characteristic differences between different members of the same cell class and how much the exact placement of the electrode (within +/-100 microns along the axon) influences the result of the simulation (Fig. 9). The membrane parameters were further examined by injection of a hyperpolarizing current pulse (Fig. 10). The resulting compartmental models (Fig. 11) based on the passive membrane parameters determined in this way form the basis of forthcoming studies on dendritic integration and signal propagation in the fly tangential cells (Haag et al., 1997; Haag and Borst, 1997).

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Cholinergic and GABAergic receptors on fly tangential cells and their role in visual motion detection.

1. To identify some of the neurotransmitters involved in the processing of visual motion information the pharmacology of transmitter receptors on motion-sensitive visual interneurons (VS and HS cells) was investigated in an in vitro preparation of the blowfly (Calliphora erythrocephala) brain. Cholinergic and GABAergic drugs were applied in the bath and iontophoretically while recording intracellularly from HS and VS cells. 2. Bath-applied carbachol (10 and 100 microM) leads to a depolarization in HS and VS cells. One micromolar nicotine also has a depolarizing effect. Both agonists are effective in 0 Ca2+/high Mg(2+)-saline, too, which isolates the cells synaptically. The muscarinic agonists pilocarpine and oxotremorine have no effects on the membrane potential. 3. Iontophoretic application of acetylcholine, carbachol, and nicotine depolarizes VS and HS cells. The iontophoretic carbachol response is antagonized by alpha-bungarotoxin (EC50 = 0.19 microM), mecamylamine (EC50 = 0.32 microM), d-tubocurarine (EC50 = 9.5 microM), and bicuculline but not by decamethonium and scopolamine. 4. Bath application of muscimol strongly hyperpolarizes VS cells in normal fly saline. The gamma-aminobutyric acid-C (GABAC)-receptor agonist cis-4-aminocrotonic acid (CACA) has no effects. The hyperpolarizing response to iontophoretic applied muscimol is present in 0 Ca2+/high Mg2+ saline as well as in Co(2+)-containing saline. The muscimol response is reduced in low chloride saline and thus chloride sensitive. The muscimol response is blocked by picrotoxinin (EC50 = 3.4 microM) but not by the GABAA receptor antagonist bicuculline. 5. Taken together the primary responses of the lobula plate tangential cells appear to be nicotinic cholinergic and GABAergic. 6. The pharmacology of natural synaptic input to VS cells was investigated by extracellular electrical stimulation of the medulla. Such evoked excitatory postsynaptic potentials (EPSPs) are blocked reversibly in 0 Ca2+/high Mg2+ saline. The nicotinic antagonists mecamylamine (1 microM) and d-tubocurarine (50-100 microM) abolish or diminish the EPSPs, respectively. 7. The pharmacological data are incorporated into a semicellular model of a visual motion detector favoring a role of lobula plate tangential cells in certain steps of visual motion processing. Cholinergic and GABAergic inputs are an ideal cellular implementation of a linear subtraction of the signals arising from local motion-sensitive elements with opposite preferred direction. Such a mechanism enhances direction-selectivity and, together with dendritic integration, increases the sensitivity of the tangential cells for wide-field motion.

Adrenergic Agonists↗

Macrophage depletion in the rat after intraperitoneal administration of liposome-encapsulated clodronate: depletion kinetics and accelerated repopulation of peritoneal and omental macrophages by administration of Freund's adjuvant.

The purpose of this study was to develop a method for the depletion of macrophages from the peritoneal cavity and the omentum of the rat. Rats received two intraperitoneal injections (at days 0 and 3) with liposome-encapsulated clodronate (dichloromethylene bisphosphonate: Cl2MBP-liposomes). This treatment resulted in complete elimination of mature tissue macrophages (ED2-positive macrophages) from the peritoneal cavity and the omentum within 2 days. The elimination included the strongly ED2-positive spindle-shaped cells of the omental membrane. Repopulation of the omental ED2-positive macrophages was not seen within the next 23 days. Whereas ED2-positive macrophages were completely depleted, few ED1-positive cells remained and repopulation of ED1-positive cells was faster. The treatment further depleted macrophages from the spleen, especially from the red pulp, parathymic lymph nodes and liver. Freund's incomplete adjuvant administered one day after the last injection of Cl2MBP-liposomes considerably accelerated repopulation in the omentum. The protocol described might be used to investigate the contribution of mature tissue macrophages to the induction of immune responses, drug metabolism and the elimination of intestinal tumours.

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Mechanisms of dendritic integration underlying gain control in fly motion-sensitive interneurons.

In the compensatory optomotor response of the fly the interesting phenomenon of gain control has been observed by Reichardt and colleagues (Reichardt et al., 1983): The amplitude of the response tends to saturate with increasing stimulus size, but different saturation plateaus are assumed with different velocities at which the stimulus is moving. This characteristic can already be found in the motion-sensitive large field neurons of the fly optic lobes that play a role in mediating this behavioral response (Hausen, 1982; Reichardt et al., 1983; Egelhaaf, 1985; Haag et al., 1992). To account for gain control a model was proposed involving shunting inhibition of these cells by another cell, the so-called pool cell (Reichardt et al., 1983), both cells sharing common input from an array of local motion detectors. This article describes an alternative model which only requires dendritic integration of the output signals of two types of local motion detectors with opposite polarity. The explanation of gain control relies on recent findings that these input elements are not perfectly directionally selective and that their direction selectivity is a function of pattern velocity. As a consequence, the resulting postsynaptic potential in the dendrite of the integrating cell saturates with increasing pattern size at a level between the excitatory and inhibitory reversal potentials. The exact value of saturation is then set by the activation ratio of excitatory and inhibitory input elements which in turn is a function of other stimulus parameters such as pattern velocity. Thus, the apparently complex phenomenon of gain control can be simply explained by the biophysics of dendritic integration in conjunction with the properties of the motion-sensitive input elements.

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A preparation of the blowfly (Calliphora erythrocephala) brain for in vitro electrophysiological and pharmacological studies.

We describe a method for the preparation and maintenance of the blowfly (Calliphora erythrocephala) brain in a recording chamber under in vitro conditions in a semi-slice configuration. Large identification neurones in the posterior part of the 3rd optic lobe (lobula plate) can be penetrated easily with microelectrodes. The so-called vertical system (VS) cells which respond to vertical image motion in vivo could be encountered best because their axons are escorted individually by specific tracheae. Fluorescent stained cells show their natural shape as being in vivo. Electrophysiological properties of the cells investigated so far, i.e., resting potential (about -40 mV) and firing properties (single rebound spikes), are comparable to recordings in intact flies. Initial pharmacological experiments on VS cells in this preparation reveal that iontophoretical application of acetylcholine and carbamylcholine results in depolarization. VS cells also respond to bath-applied nicotine (1 microM) with a slow depolarization of their membrane potential in normal fly saline as well as in a Ca(2+)-free saline, suggesting direct cholinergic input via nicotinic receptors. The suitability of the preparation for a wide range of electrophysiological and pharmacological studies is discussed.

Acetylcholine↗

Calcium accumulation in visual interneurons of the fly: stimulus dependence and relationship to membrane potential.

1. The large motion-sensitive tangential neurons in the fly third visual neuropil spatially pool the postsynaptic signals of many local elements. The changes in membrane potential and calcium concentration induced in these cells by visual motion are analyzed in vivo by simultaneous optical and intracellular voltage recording techniques. 2. Visual motion in the preferred direction leads to depolarization of the cell and to calcium accumulation mainly in the axon terminal, the soma, and the dendritic tree. During motion in the null direction, the cell hyperpolarizes and virtually no changes in calcium concentration can be observed. 3. Dendritic calcium accumulation is first restricted to those dendritic branches that are close to the sites of direct synaptic input. In other parts of the dendrite the calcium concentration increases more slowly and usually reaches only lower levels. 4. Calcium starts accumulating at the onset of motion. However, the calcium concentration reaches its final steady-state level much later than the corresponding membrane potential changes. Even if these are completely transient at high temporal frequencies of pattern motion, the calcium signal stays high until the stimulus pattern stops moving. 5. The amplitude of the calcium signal depends on the temporal frequency of pattern motion in a similar way as do the corresponding membrane potential changes. However, there exist differences that can be attributed to the different time courses of both signals. 6. Depolarization of the dendritic tree by current injection through a microelectrode leads to similar changes in calcium accumulation as does activation by synaptic input, suggesting that calcium enters the cell via voltage-dependent channels. The possible function of calcium channels for dendritic integration of synaptic input is discussed.

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Processing of synaptic information depends on the structure of the dendritic tree.

The consequences of dendritic geometry for the processing of synaptic information was analysed in two types of motion-sensitive neurones in the visual system of the fly. These neurones differ conspicuously in the morphology of their dendrites but receive their input from the same type of local motion-sensitive elements. Intracellular recording and activating selected regions of the dendrite by visual motion showed that, in accordance with cable theory of nerve cells, the way in which postsynaptic signals interact is essentially determined by the structure of the dendritic tree.

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Dendritic processing of synaptic information by sensory interneurons.

One of the most distinguishing features of nerve cells is the vast morphological diversity of their input regions, that is, their dendrites. These range from bulbous structures, with only small protrusions, to large tree-like arborizations. The diversity of nerve cells is further augmented by a continuously increasing number of types of voltage-dependent conductances in dendrites that might alter the postsynaptic signals in a pronounced way. Moreover, intracellular factors such as Ca2+ link electrical activity with biochemical processes, and can induce short and long-term changes in responsiveness. This complexity of neurons in general, and the uniqueness of each cell type, sharply contrasts with the comparatively simple and uniform design principle of the integrate-and-fire units of so-called neuronal net models. This raises the question of which particular structural and physiological details of nerve cells really matter for the performance of neuronal circuits. An answer to this basic problem of computational neurobiology might be given only if the task of the neurons and circuits is known. This review illustrates how the problem can be approached particularly well in sensory interneurons. The functional significance of sensory interneurons can often be assessed more easily than that of central nerve cells because of their vicinity to the sensory surface.

Animals↗

Motion computation and visual orientation in flies.

1. Visual orientation greatly relies on the evaluation of the motion patterns received by the eyes when the animal moves around. 2. In a combination of behavioral, neurophysiological and pharmacological analysis and modelling, the mechanisms are established by which the visual system of the fly extracts three types of-basic retinal motion patterns. 3. Coherent retinal large-filed motion as is induced during deviations of the animal from its course, image expansion occurring when the animal approaches an obstacle, and relative motion which is induced when a nearby object is passed in front of its background. 4. Separate neuronal networks are specifically tuned to each of these motion patterns and make use of them in three different orientation tasks: in compensatory course stabilization, the control of landing behaviour and the fixation of objects.

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Neural circuit tuning fly visual interneurons to motion of small objects. I. Dissection of the circuit by pharmacological and photoinactivation techniques.

1. Visual interneurons tuned to the motion of small objects are found in many animal species and are assumed to be the neuronal basis of figure-ground discrimination by relative motion. A well-examined example is the FD1-cell in the third visual neuropil of blowflies. This cell type responds best to motion of small objects. Motion of extended patterns elicits only small responses. As a neuronal mechanism that leads to such a response characteristic, it was proposed that the FD1-cell is inhibited by the two presumably GABAergic and, thus, inhibitory CH-cells, the VCH- and the DCH-cell. The CH-cells respond best to exactly that type of motion by which the activity of the FD1-cell is reduced. The hypothesis that the CH-cells inhibit the FD1-cell and, thus, mediate its selectivity to small moving objects was tested by ablating the CH-cells either pharmacologically or by photoinactivation. 2. After application of the gamma-aminobutyric acid (GABA) antagonist picrotoxinin, the FD1-cell responds more strongly to large-field than to small-field motion, i.e., it has lost its small-field selectivity. This suggests that the tuning of the FD1-cell to small moving objects relies on a GABAergic mechanism and, thus, most likely on the CH-cells. 3. The role of each CH-cell for small-field tuning was determined by inactivating them individually. They were injected with a fluorescent dye and then ablated by laser illumination. Only photoinactivation of the VCH-cell eliminated the specific selectivity of the FD1-cell for small-field motion. Ablation of the DCH-cell did not significantly change the response characteristic of the FD1-cell. This reveals the important role of the VCH-cells in mediating the characteristic sensitivity of the FD1-cell to motion of small objects. 4. The FD1-cell is most sensitive to motion of small objects in the ventral part of the ipsilateral visual field, whereas motion in the dorsal part influences the cell only weakly. This specific feature fits well to the sensitivity of the VCH-cell to ipsilateral motion that is most pronounced in the ventral part of the visual field. The spatial sensitivity distribution of the FD1-cell matches also the characteristics of figure-ground discrimination and fixation behavior.

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