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A Borst

Publications and source records attributed to A Borst.

At least 19 recordsLinked to original sources

Local current spread in electrically compact neurons of the fly.

Analyses of active and passive membrane properties predict an asymmetry in the spread of electrical current through a neuron. Simulated current injection into a large-diameter compartment of a biophysically realistic model neuron causes a local potential shift that can spread throughout the cell. In contrast, causing the same local potential shift in the dendritic tip of the same neuron results in only minimal changes in electrical potential in the rest of the cell. Using calcium as a reporter of electrical activity in neurons in the fly's lobula plate we find that current injected into the thick axon caused depolarization throughout the cell, whereas activation of a dendritic region remained local. These results have important implications for the ability of integrating neurons to perform local computations of synaptic input without additional hardware.

Animals↗

Spatial distribution and characteristics of voltage-gated calcium signals within visual interneurons.

Most of our knowledge about insect calcium currents is derived from studies on cultured or dissociated somata. So far, only little data on calcium currents are available for neurons including their dendritic and presynaptic structures. Here we combined the switched-electrode voltage-clamp technique with optical recording using calcium-sensitive dyes in identified fly visual interneurons in vivo to characterize the voltage dependence and dynamics of calcium currents quantitatively and in a spatially resolved way. For all three cell types considered, i.e., centrifugal horizontal (CH), horizontal system (HS), and vertical system (VS) cells, the activation curve is rather flat and covers a voltage range from -60 to -20 mV in dendritic as well as presynaptic areas of the cells. The calcium increase is fastest for CH cells with a time constant of approximately 70 ms. In HS and VS cells, the time constant amounts to 400-700 ms. The calcium dynamics as determined in different regions of the cells are similar except for a small segment between the axon and the dendrite in HS and VS cells, where the calcium increase is significantly faster. In summary, the results show the existence of a low-voltage-activated calcium current with little or no inactivation in dendritic as well as presynaptic regions of fly lobula plate tangential cells.

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Separation of voltage- and ligand-gated calcium influx in locust neurons by optical imaging.

Calcium ions can enter neurons through either ionotropic transmitter receptors or through voltage-gated calcium channels. Thus, an observed rise in intracellular calcium concentration upon synaptic stimulation can be due to either one of these mechanisms or to both of them. We analyzed the individual contribution of transmitter- and voltage-gated calcium entry in non-spiking somata, acutely dissociated from thoracic ganglia of the locust Locusta migratoria. By optically recording the calcium signal following different stimulation protocols, we isolated the voltage- and the transmitter-gated component and found that these components indeed summate to the total rise in calcium observed under control conditions.

Action Potentials↗

Spatial response properties of contralateral inhibited lobula plate tangential cells in the fly visual system.

This study describes the spatial response properties of a particular group of motion-sensitive and directionally selective neurons located in the lobula plate of the fly visual system. Their preferred motion direction is front-to-back (depolarization), and their null direction is back-to-front (hyperpolarization). They receive inhibitory input from the contralateral eye during pattern motion from back to front (regressive). In this study, we call these neurons regressive contralateral inhibited lobula plate tangential cells (rCI-LPTCs). Three physiologic groups of rCI-neurons (rCI-I, rCI-IIa, and rCI-IIb) can be distinguished on the basis of their ipsilateral pattern size dependence and their inhibitory contralateral input. rCI-I neurons depolarize during the motion of small ipsilateral patterns from front to back, but they become hyperpolarized by large ipsilateral patterns moving in the same direction. rCI-IIa and rCI-IIb neurons receive bidirectional inhibitory input from the contralateral eye. rCI-IIa neurons respond best to small ipsilateral pattern sizes, but unlike rCI-I neurons, their net response to large patterns is positive. rCI-IIb neurons respond best to large ipsilateral patterns. The anatomical and physiologic variability of the rCI-neurons suggests that more than three types of rCI-neurons exist. The suggested physiologic groups might be preliminary. We recorded one neuron that could mediate the bidirectional inhibitory input that rCI-IIa and rCI-IIb neurons receive from the contralateral eye. In the case of the rCI-IIa neurons at least one further contralateral inhibitory element has to be assumed. The tuning of rCI-IIa neurons to small ipsilateral pattern sizes is likely to be based on an on-center/off-surround organization of their synaptic input.

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Nucleic acid sequence-based amplification (NASBA) detection of medically important Candida species.

Nucleic Acid Sequence Based Amplification (iNASBA), an isothermal amplification technique for nucleic acids, was evaluated for the identification of medically important Candida species using primers selected from 18S rRNA sequences conserved in fungi. An RNA fragment of 257 nucleotides was amplified for Candida albicans. Nineteen different fungi were tested for rRNA amplification with the NASBA. All were positive when analyzed on agarose gel, whereas human RNA was negative. For the identification of Candida species, NASBA amplification products were analyzed in an enzyme bead-based detection format, using species-specific biotinylated probes and a generic Candida HRPO probe or a membrane-based system using biotinylated probes and avidin-HPRO. Discrimination of the major human pathogenic Candida spp. was based on a panel of biotinylated probes for C. krusei, C. tropicalis, C. albicans, C. glabrata, and C. lusitaniae. Using rRNA dilutions obtained from pure cultures of C. albicans, the combination of NASBA and the enzymatic bead-based detection yielded a sensitivity equivalent to 0.01 CFU. In a model system using 1 ml of artificially contaminated blood as few as 1-10 CFU of C. albicans could be detected. Testing of 68 clinical blood samples from patients suspected of candidemia showed that eight samples were positive for C. albicans and one for C. glabrata. Testing of 13 clinical plasma samples from patients suspected of fungemia identified the presence of C. albicans in two specimens. The whole procedure of sample preparation, amplification and identification by hybridization can be performed in 1 day. This speed and the observed sensitivity of the assay make the NASBA a good alternative to PCR for the detection of candidemia.

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Information theory and neural coding.

Information theory quantifies how much information a neural response carries about the stimulus. This can be compared to the information transferred in particular models of the stimulus-response function and to maximum possible information transfer. Such comparisons are crucial because they validate assumptions present in any neurophysiological analysis. Here we review information-theory basics before demonstrating its use in neural coding. We show how to use information theory to validate simple stimulus-response models of neural coding of dynamic stimuli. Because these models require specification of spike timing precision, they can reveal which time scales contain information in neural coding. This approach shows that dynamic stimuli can be encoded efficiently by single neurons and that each spike contributes to information transmission. We argue, however, that the data obtained so far do not suggest a temporal code, in which the placement of spikes relative to each other yields additional information.

Algorithms↗

Active membrane properties and signal encoding in graded potential neurons.

We investigated the influence of active membrane properties on the precision by which the stimulus velocity is encoded in the membrane potential of a motion-sensitive interneuron in the blowfly. The so-called HS-cells respond to visual motion stimuli with a graded shift in membrane potential. Superimposed on this graded response are small spike-like events. This "mixed" visual response mode can be modified by current injection in two different ways. (1) By ongoing injection of hyperpolarizing current, the spike-like events are turned into full-blown action potentials, and (2) by injection of depolarizing current, the spike-like events become completely suppressed. The visual response then consists of a graded shift of membrane potential only. As a measure of the fidelity, we calculated the coherence between the motion stimulus and the response of the cell elicited with different electrical manipulations of the cell. We found that the coherence was highest for the cell at rest. Any electrical manipulation resulted in a reduced coherence. This was attributable partly to a lower signal-to-noise ratio and partly to an increased nonlinearity in the response. By applying a threshold operation we transformed the analog membrane response into an all-or-none spike train. A comparison between these two ways of signal representation revealed that more information about the stimulus velocity is inherent in the analog membrane potential than in the spike train.

Action Potentials↗

Dendritic integration and its role in computing image velocity.

The mechanisms underlying visual motion detection can be studied simultaneously in different cell compartments in vivo by using calcium as a reporter of the spatiotemporal activity distribution in single motion-sensitive cells of the fly. As predicted by the Reichardt model, local dendritic calcium signals are found to indicate the direction and velocity of pattern motion but are corrupted by spatial pattern properties. The latter are canceled out by spatial integration, thus leading to a purely directional selective output signal in the axon. These findings attribute a specific computational task to the dendrites of visual interneurons and imply a functional interpretation of dendritic morphology.

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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.

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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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