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B Barbour

Publications and source records attributed to B Barbour.

At least 19 recordsLinked to original sources
PubMed2001

An evaluation of synapse independence.

If, as is widely believed, information is stored in the brain as distributed modifications of synaptic efficacy, it can be argued that the storage capacity of the brain will be maximized if the number of synapses that operate independently is as large as possible. The majority of synapses in the brain are glutamatergic; their independence will be compromised if glutamate released at one synapse can significantly activate receptors at neighboring synapses. There is currently no agreement on whether "spillover" after the liberation of a vesicle will significantly activate receptors at neighboring synapses. To evaluate the independence of central synapses, it is necessary to compare synaptic responses with those generated at neighboring synapses by glutamate spillover. Here, synaptic activation and spillover responses are simulated in a model, based on data for hippocampal synapses, that includes an approximate representation of the extrasynaptic space. Recently-published data on glutamate transporter distribution and properties are incorporated. Factors likely to influence synaptic or spillover responses are investigated. For release of one vesicle, it is estimated that the mean response at the nearest neighboring synapse will be <5% of the synaptic response. It is concluded that synapses can operate independently.

ATP-Binding Cassette Transporters↗
PubMed2001

Functional antigen-independent synapses formed between T cells and dendritic cells.

Immunological synapse formation is usually assumed to require antigen recognition by T cell receptors. However, the immunological synapse formed at the interface between naïve T cells and dendritic cells (DCs) has never been described. We show here that in the absence of antigen, and even of major histocompatibility complex molecules, T cell-DC synapses are formed and lead to several T cell responses: a local increase in tyrosine phosphorylation, small Ca2+ responses, weak proliferation and long-term survival. These responses are triggered more readily in CD4+ T cells than in CD8+ T cells, which express a specific isoform of the repulsive molecule CD43. These phenomena may play a major role in the maintenance of the naïve T cell pool in vivo.

Animals↗
PubMed2000

Combining loose cell-attached stimulation and recording.

Many synaptic-physiology experiments require selective stimulation of presynaptic neurones. No single method for achieving this is entirely satisfactory. Presynaptic whole-cell or microelectrode recordings offer outstanding control, but establishing such recordings of synaptically-connected neurones can be very time consuming. Minimal stimulation provides too little information regarding the condition and excitation of the presynaptic neurone(s). In the loose cell-attached configuration, it is possible both to stimulate individual cells and to record their action potentials, but most commercially-available equipment will not enable combining stimulation and recording. We demonstrate simultaneous loose cell-attached stimulation and recording of action potentials using a specially-designed patch-clamp amplifier. Since no tight seal is required, the method permits electrode re-use, thus enabling rapid screening of presynaptic cells while stimulating them selectively.

Action Potentials↗
PubMed1997

Currents evoked in Bergmann glial cells by parallel fibre stimulation in rat cerebellar slices.

1. Whole-cell recordings were obtained from Bergmann glial cells in rat cerebellar slices. 2. The cells had low input resistances (70 +/- 38 M omega; n = 13) and a mean resting potential of -82 +/- 6 mV (n = 12) with a potassium-based internal solution. Electrical and dye coupling between Bergmann glia were confirmed. 3. Stimulation of parallel fibres induced a complex, mostly inward current which could be decomposed pharmacologically. 4. The ionotropic glutamate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM), but not DL-2-amino-5-phosphonopentanoic acid (DL-APV; 100 microM) consistently blocked an early inward current component that may reflect synaptic activation of AMPA/kainate receptors in Bergmann glia. 5. Addition of cadmium ions (100 microM) to inhibit transmitter release blocked most of the CNQX-APV-insensitive current. This component probably reflects electrogenic uptake of the synaptically released glutamate. 6. Tetrodotoxin (TTX; 1 microM) blocked the remaining inward current: a slow component, possibly produced by the potassium ion efflux during action potential propagation in parallel fibres. An initial triphasic component of the response was also TTX sensitive and reflected passage of the parallel fibre action potential volley. 7. The putative glutamate uptake current was further characterized; it was blocked by the competitive uptake blockers D-aspartate (0.5 mM) and L-trans-pyrrolidine-2,4-dicarboxylic acid (PDC; 0.5 mM), and by replacement of sodium with lithium. Monitoring the triphasic TTX-sensitive component showed that this inhibition did not result from changes of action potential excitation and propagation. 8. Intracellular nitrate ions increased the putative uptake current, consistent with the effect of this anion on glutamate transporters. 9. The putative uptake current was reduced by depolarization, consistent with the voltage dependence of glutamate uptake. 10. It is concluded that a large fraction of the current induced by parallel fibre stimulation reflects the uptake of synaptically released glutamate. The uptake current activated rapidly, with a 20-80% rise time of 2.3 +/- 0.7 ms (n = 10), and decayed with a principal time constant of 25 +/- 6 ms (n = 10).

2-Amino-5-phosphonovalerate↗
PubMed1997

Intersynaptic diffusion of neurotransmitter.

According to many theories of brain function, the computational power of the brain depends upon the number of independent synapses it contains. A synapse will not be independent if its receptors are activated or modified by neurotransmitter released at neighbouring synapses. Recently, there have been several reports suggesting the occurrence of 'crosstalk' or 'spillover', and a large number of results consistent with crosstalk. However, the quantitative importance of this phenomenon remains uncertain. We estimate the significance of crosstalk using a simple model which predicts that, during concentrated synaptic activity, crosstalk between distinct synapses is likely to activate high-affinity receptors and may also desensitize certain receptors. Comparison of these predictions with the experimental data highlights the information that is required for a more detailed model of crosstalk.

Animals↗
PubMed1994

Counter-transport of potassium by the glutamate uptake carrier in glial cells isolated from the tiger salamander retina.

1. To investigate the transport of potassium on the glutamate uptake carrier, the glutamate uptake current in isolated retinal Müller cells was monitored by whole-cell clamping, while measuring changes of potassium concentration outside the cells ([K+]o) with an ion-sensitive microelectrode. 2. Activating glutamate uptake led to an accumulation of potassium outside the cells, consistent with the hypothesis, based on less direct evidence, that the glutamate uptake carrier transports potassium out of the cell. 3. The glutamate-evoked rise of [K+]o showed the pharmacology and sodium dependence of glutamate uptake. 4. The rise in [K+]o was proportional to the uptake current flowing between 0 and -80 mV, implying that the ratio of K+ transported to charge transported by the uptake carrier is constant over this voltage range. The K+ to charge transport ratio was the same for uptake of D-aspartate and L-glutamate. 5. By clamping cells with pipettes containing solutions of different [K+], the dependence of the glutamate and aspartate uptake currents on intracellular [K+] was determined. L- and D-aspartate transport showed a smaller maximum uptake current (Imax), and a smaller apparent Michaelis constant (Km) for activation by intracellular K+, than did L-glutamate transport. The ratio of Imax to Km was the same for these three analogues, a result which can be predicted from simple models of the carrier's operation. 6. Fully activating glutamate uptake in Müller cells in the intact retina would produce a K+ load into the extracellular space of about 0.6 mM s-1. Suppression of glutamate release from photoreceptors by light will reduce K+ efflux from Müller cells in the outer retina; this may contribute to the light-evoked fall of [K+]o observed in the outer retina, and thus contribute to shaping the electroretinogram.

Amino Acid Transport System X-AG↗
PubMed1994

Prolonged presence of glutamate during excitatory synaptic transmission to cerebellar Purkinje cells.

In the molecular layer of the cerebellar cortex, Purkinje cells and interneurons receive a common excitatory input from parallel fibers. The AMPA/kainate receptor-mediated parallel fiber excitatory postsynaptic current (EPSC) recorded in Purkinje cells decays much more slowly than that recorded in interneurons. We show that this slowness of decay does not result from dendritic filtering and that it is unlikely to reflect the deactivation kinetics of the postsynaptic receptors. Agents blocking glutamate uptake prolong the EPSC in Purkinje cells. We conclude that the slow EPSC decay results from the continued presence of transmitter glutamate. This may be due to retarded transmitter diffusion around spines or to cross-talk between neighboring active synapses.

Animals↗
PubMed1993

Synaptic currents evoked in Purkinje cells by stimulating individual granule cells.

Each cerebellar Purkinje cell receives input from about 160,000 glutamatergic granule cells. In anatomically intact preparations this input has hitherto been studied only as a compound synaptic potential or current. Presented here are simultaneous recordings in cerebellar slices of synaptically connected granule cell-Purkinje cell pairs. The mean amplitude of the excitatory synaptic currents evoked by stimulation of individual granule cells ranged from 2 to 60 pA, whereas the great majority of the spontaneous glutamatergic currents in Purkinje cells in the presence of tetrodotoxin were < 40 pA. In several cases, stimulation of a single granule cell evoked a disynaptic inhibitory current. It is estimated that on the order of 50 simultaneously active granule cells are sufficient to excite a Purkinje cell.

Animals↗
PubMed1993

Changes in NAD(P)H fluorescence and membrane current produced by glutamate uptake into salamander Müller cells.

1. Glutamate uptake into isolated, whole-cell patch-clamped glial cells was studied by monitoring the increase of cell fluorescence generated as glutamate and NAD(P) were converted into alpha-ketoglutarate and NAD(P)H by glutamate dehydrogenase. The current generated by the glutamate uptake carrier was recorded simultaneously. 2. L-Glutamate evoked an increase of cell fluorescence and an inward uptake current. L- and D-aspartate generated an uptake current but no fluorescence response, consistent with the amino acid specificity of glutamate dehydrogenase. 3. In the absence of external sodium the glutamate-evoked fluorescence response and uptake current were abolished, showing that there is no sodium-independent glutamate uptake across the cell membrane. 4. Varying the glutamate concentration altered both the fluorescence response and the uptake current. The fluorescence response saturated at a lower glutamate concentration than the uptake current, and depended in a Michaelis-Menten fashion on the uptake current. 5. The fluorescence response and the uptake current were reduced by membrane depolarization, and also by removal of intracellular potassium. 6. The dependence of the fluorescence response on uptake current when membrane potential was altered or intracellular potassium was removed was the same as that seen when the external glutamate concentration was altered. 7. These fluorescence studies show that glutamate uptake is inhibited by depolarization and by removal of intracellular potassium, consistent with the conclusion of earlier work in which uptake was monitored solely as a membrane current. The data are consistent with high-affinity electrogenic sodium- and potassium-dependent glutamate uptake with fixed stoichiometry being the only significant influx route for glutamate. Other possible interpretations of the data are also discussed.

Action Potentials↗
PubMed1991

The potassium-dependence of excitatory amino acid transport: resolution of a paradox.

Schwartz and Tachibana have claimed that the uptake of excitatory amino acids (EAAs) into glia is independent of intracellular potassium, in contradiction to the results of our previous studies. We show here that failure to observe the potassium-dependence of uptake resulted from the use of small whole-cell pipettes, which fail to dialyze properly the cell interior, and the use of D-aspartate instead of L-glutamate as the transported EAA, the uptake of D-aspartate being maintained down to lower levels of intracellular potassium than is the uptake of L-glutamate.

Amino Acids↗
PubMed1991

Electrogenic uptake of glutamate and aspartate into glial cells isolated from the salamander (Ambystoma) retina.

The effects of excitatory amino acids on the membrane current of isolated retinal glial cells (Müller cells) were investigated using whole-cell patch clamping. 2. L-Glutamate evoked an inward current at membrane potentials between -140 and +50 mV. The current was larger at more negative potentials. 3. The glutamate-evoked current was activated by external cations with relative efficacies: Na+ much greater than Li+ greater than K+ greater than Cs+, choline. It was activated by internal cations with relative efficacies K+ greater than Rb+ greater than Cs+ much greater than choline. Chloride and divalent cations did not affect the glutamate-evoked current. 4. Raising the intracellular sodium or glutamate concentrations, or raising the extracellular potassium concentration, reduced the current evoked by external glutamate. The suppressive effect of internal glutamate was larger when the internal sodium concentration was high. 5. Some analogues of glutamate also evoked an inward current. Responses to L-aspartate resembled those to glutamate, but for aspartate the apparent affinity was higher and the voltage dependence of the current was steeper. In the physiological potential range the current evoked by a saturating dose of aspartate was less than that evoked by a saturating dose of glutamate. 6. The uptake blocker threo-3-hydroxy-DL-aspartate (30 microM) reduced the glutamate-evoked current, but also generated a current itself. Dihydrokainate (510 microMs) weakly inhibited the glutamate-evoked current without generating a current itself. 7. The commonly used blockers of glutamate-gated ion channels, 2-amino-5-phosphonovalerate (APV; 100 microMs), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 20 microMs), and kynurenate (1mM) had no effect on the glutamate-evoked current. 8. The voltage dependence, cation dependence and pharmacological profile of the current evoked by excitatory amino acids indicate that it is caused by activation of the high-affinity glutamate uptake carrier. This carrier appears to transport one glutamate anion into the cell, one K+ ion out of the cell, and two or more Na+ ions into the cell, on each carrier cycle. At the inner membrane surface some or all of the transported Na+ dissociates from the carrier after the transported glutamate has dissociated. 9. In addition to glutamate, the uptake carrier can also transport aspartate and threo-3-hydroxy-DL-aspartate, but not dihydrokainate.

Ambystoma↗
PubMed1990

Non-vesicular release of glutamate from glial cells by reversed electrogenic glutamate uptake.

Glutamate uptake into nerve and glial cells usually functions to keep the extracellular glutamate concentration low in the central nervous system. But one component of glutamate release from neurons is calcium-independent, suggesting a non-vesicular release that may be due to a reversal of glutamate uptake. The activity of the electrogenic glutamate uptake carrier can be monitored by measuring the membrane current it produces, and uptake is activated by intracellular potassium ions. Here we report that raising the potassium concentration around glial cells evokes an outward current component produced by reversed glutamate uptake. This current is activated by intracellular glutamate and sodium, inhibited by extracellular glutamate and sodium, and increased by membrane depolarization. These results demonstrate a non-vesicular mechanism for the release of glutamate from glial cells and neurons. This mechanism may contribute to the neurotoxic rise in extracellular glutamate concentration during brain anoxia.

Animals↗
PubMed1988

Electrogenic glutamate uptake in glial cells is activated by intracellular potassium.

Uptake of glutamate into glial cells in the CNS maintains the extracellular glutamate concentration below neurotoxic levels and helps terminate its action as a neurotransmitter. The co-transport of two sodium ions on the glutamate carrier is thought to provide the energy needed to transport glutamate into cells. We have shown recently that glutamate uptake can be detected electrically because the excess of Na+ ions transported with each glutamate anion results in a net current flow into the cell. We took advantage of the control of the environment, both inside and outside the cell, provided by whole-cell patch-clamping and now report that glutamate uptake is activated by intracellular potassium and inhibited by extracellular potassium. Our results indicate that one K+ ion is transported out of the cell each time a glutamate anion and three Na+ ions are transported in. A carrier with this stoichiometry can accumulate glutamate against a much greater concentration gradient than a carrier co-transporting one glutamate anion and two Na+ ions. Pathological rises in extracellular potassium concentration will inhibit glutamate uptake by depolarizing glial cells and by preventing the loss of K+ from the glutamate carrier. This will facilitate a rise in the extracellular glutamate concentration to neurotoxic levels and contribute to the neuronal death occurring in brain anoxia and ischaemia.

Animals↗