Input Conductance Research Articles

Regulation of the subthreshold chloride conductance in the rat sympathetic neuron

The mechanisms that control chloride conductance (gCl) in the rat sympathetic neuron have been studied by the two-electrode voltage-clamp technique in mature, intact superior cervical ganglia in vitro. In addition to voltage dependence in the membrane potential range -120/-50 mV, gCl displays time- and activity-dependent regulation (sensitization). The resting membrane potential is governed by voltage-dependent gK and gCl, which determine values of cell input conductance ranging from 7 to 18 nS (full deactivation) to an upper value of about 130 nS (full activation and maximal gCl sensitization). The quiescent neuron, held at constant membrane potential, spontaneously and gradually moved from a low- to a high-conductance status. An increase (about 40 nS) in gCl accounted for this phenomenon, which could be prevented by imposing intermittent hyperpolarizing episodes. Following spike firing, gCl increased by 20-33 nS, independent of the cell conductance value preceding tetanization, and thereafter decayed to the pre-stimulus level within 5 min. Intracellular sodium depletion and its successive ionophoretic restoration moved the neuron from a stable low-conductance state to maximum gCl sensitization, pointing to a link between gCl sensitization and [Na+]i. The dependence of gCl build-up on [Na+]i and the time-course of such Na+-related modulation have been examined: gCl sensitization was absent at 0 [Na+]i, was well developed (20 nS) at 15 mM and tended towards a saturating value of 60 nS for higher [Na+]i. Sensitization was transient in response to neuron activity. In the silent neuron, sensitization of gCl shifted membrane potential over a range of about 15 mV.

Barbiturate activation and modulation of GABA A receptors in neocortex

We determined if anesthetic and anti-epileptic barbiturates inhibit neurons by different mechanisms. Current- and voltage-clamp recordings were made from somatosensory neurons of neocortex and some thalamocortical neurons in coronal brain slices of rats. We compared effects of pentobarbital, amobarbital, and phenobarbital on inhibitory postsynaptic currents (IPSCs) mediated by γ-aminobutyric acid (GABA), input conductance, and evoked action potential firing. In neocortex, pentobarbital (EC 50 = 41 μM) and amobarbital (EC 50 = 103 μM) increased the decay time constant of GABA Aergic IPSCs. At higher concentrations, pentobarbital and amobarbital shunted firing by increasing input conductance through agonism at GABA A receptors. At anti-epileptic concentrations, phenobarbital increased the IPSC decay time constant (EC 50 = 144 μM), and shunted firing by agonism at GABA A receptors (EC 50 = 133 μM). In thalamocortical neurons, similar concentrations of phenobarbital had negligible effects on GABA Aergic IPSCs, conductance, and firing. In contrast to their thalamic actions, barbiturates inhibit neocortical neurons mostly through GABA receptors. Neocortical enhancement of inhibition by pentobarbital and amobarbital, combined with actions on thalamocortical neurons, may contribute to redundant mechanisms of anesthesia. The ability of phenobarbital at anti-epileptic concentrations to inhibit neocortical firing by direct activation and modulation of GABA A receptors relates to its specialized therapeutic effects.

Role for the Subthreshold Currents ILeak and IH in the Homeostatic Control of Excitability in Neocortical Somatostatin-Positive Inhibitory Neurons

Cortical circuitry reconfigures in response to chronic (1-3 days) changes in activity levels. To understand this process, we must know the role played by inhibitory neurons because they crucially influence network properties by controlling action potential generation and synaptic integration. Using pharmacological blockade of activity in neocortical organotypic slice cultures, we examined the activity-dependent regulation of membrane excitability in a specific inhibitory neuron subtype: the somatostatin-positive (SOM+) neuron. Chronic action potential blockade (TTX, 2.5 days) resulted in increased excitability in SOM+ neurons. This result is consistent with a homeostatic process to maintain the average firing rate of SOM+ neurons at a particular level. Excitability changes were not ascribed to changing cell size or alterations in voltage-dependent sodium current. Instead, the excitability increase was largely the result of a decrease in the density of two subthreshold currents: a passive leak current (ILeak) and H-current (IH). The downregulation of these currents increased excitability mostly through a decrease in membrane input conductance. The coadaptation of ILeak and IH enabled a change in input conductance while helping to preserve membrane potential. Evidence indicated that ILeak was probably mainly mediated by K+. At earlier culture ages, this adaptation was superimposed on developmental changes, whereas at older ages, the same types of induced alterations occurred but with no developmental component. Together with other studies, these data indicate that both inhibitory and excitatory neurons increase membrane excitability with chronic reduction in activity, but through different mechanisms.

Suprachiasmatic nucleus communicates with anterior thalamic paraventricular nucleus neurons via rapid glutamatergic and gabaergic neurotransmission: State-dependent response patterns observed in vitro

The hypothalamic suprachiasmatic nucleus uniquely projects to the midline thalamic paraventricular nucleus. To characterize this projection, patch clamp techniques applied in acute rat brain slice preparations examined responses of anterior thalamic paraventricular nucleus neurons to focal suprachiasmatic nucleus stimulation. Whole cell recordings from slices obtained during daytime (n=40) revealed neurons with a mean membrane potential of −66±1.2 mV, input conductance of 1.5±0.1 nS and state-dependent tonic or burst firing patterns. Electrical stimulation (one or four pulses) in suprachiasmatic nucleus elicited monosynaptic excitatory postsynaptic potentials (mean latency of 12.6±0.6 ms; n=12), featuring both AMPA and N-methyl-d-aspartate–glutamate receptor-mediated components, and monosynaptic bicuculline-sensitive inhibitory postsynaptic potentials (mean latency of 16.6±0.6 ms; n=7) reversing polarity at −72±2.6 mV, close to the chloride equilibrium potential. Glutamate microstimulation of suprachiasmatic nucleus also elicited transient increases in spontaneous excitatory or inhibitory postsynaptic currents in anterior thalamic paraventricular neurons. Recordings from rats under reverse light/dark conditions (n=22) yielded essentially similar responses to electrical stimulation. At depolarized membrane potentials, suprachiasmatic nucleus-evoked excitatory postsynaptic potentials triggered single action potentials, while evoked inhibitory postsynaptic potentials elicited a silent period in ongoing tonic firing. By contrast, after manual adjustment of membrane potentials to hyperpolarized levels, neuronal response to the same ‘excitatory’ stimulus was a low threshold spike and superimposed burst firing, while responses to ‘inhibitory’ stimuli paradoxically elicited excitatory rebound low threshold spikes and burst firing. These data support the existence of glutamatergic and GABAergic efferents from the suprachiasmatic nucleus to its target neurons. Additionally, in thalamic paraventricular nucleus neurons, responses to activation of their suprachiasmatic afferents may vary in accordance with their membrane potential–dependent intrinsic properties, a characteristic typical of thalamocortical neurons.

Modifying the Subunit Composition of TASK Channels Alters the Modulation of a Leak Conductance in Cerebellar Granule Neurons

Two-pore domain potassium (K2P) channel expression is believed to underlie the developmental emergence of a potassium leak conductance [IK(SO)] in cerebellar granule neurons (CGNs), suggesting that K2P function is an important determinant of the input conductance and resting membrane potential. To investigate the role that different K2P channels may play in the regulation of CGN excitability, we generated a mouse lacking TASK-1, a K2P channel known to have high expression levels in CGNs. In situ hybridization and real-time PCR studies in wild-type and TASK-1 knock-outs (KOs) demonstrated that the expression of other K2P channels was unaltered in CGNs. TASK-1 knock-out mice were healthy and bred normally but exhibited compromised motor performance consistent with altered cerebellar function. Whole-cell recordings from adult cerebellar slice preparations revealed that the resting excitability of mature CGNs was no different in TASK-1 KO and littermate controls. However, the modulation of IK(SO) by extracellular Zn2+, ruthenium red, and H+ was altered. The IK(SO) recorded from TASK-1 knock-out CGNs was no longer sensitive to alkalization and was blocked by Zn2+ and ruthenium red. These results suggest that a TASK-1-containing channel population has been replaced by a homodimeric TASK-3 population in the TASK-1 knock-out. These data directly demonstrate that TASK-1 channels contribute to the properties of IK(SO) in adult CGNs. However, TASK channel subunit composition does not alter the resting excitability of CGNs but does influence sensitivity to endogenous modulators such as Zn2+ and H+.

How Membrane Properties Shape the Discharge of Motoneurons: A Detailed Analytical Study

Electrophysiological experiments and modeling studies have shown that afterhyperpolarization regulates the discharge of lumbar motoneurons in anesthetized cats and is an important determinant of their firing properties. However, it is still unclear how firing properties depend on slow afterhyperpolarization, input conductance, and the fast currents responsible for spike generation. We study a single-compartment integrate-and-fire model with a slow potassium conductance that exponentially decays between spikes. We show that this model is analytically solvable, and we investigate how passive and active membrane properties control the discharge. We show that the model exhibits three distinct firing ranges (primary, secondary, and high frequency), and we explain the origin of these three ranges. Explicit expressions are established for the gain of the steady-state current-frequency (I-f) curve in the primary range and for the gain of the I-f curve for the first interspike interval. They show how the gain is controlled by the maximal conductance and the kinetic parameters of the afterhyperpolarization conductance. The gain also depends on the spike voltage threshold, and we compute how it is decreased by threshold accommodation (i.e., the increase of the threshold with the injected current). In contrast, the gain is not very sensitive to the input conductance. This implies that tonic synaptic activity shifts the current-frequency curve in its primary range, in agreement with experiments. Taking into account the absolute refractory period associated with spikes somewhat reduces the gain in the primary range. More importantly, it accounts for the approximately linear and steep secondary range observed in many motoneurons. In the nonphysiological high-frequency range, the behavior of the I-f curve is determined by the fast voltage-dependent currents, via the amplitude of the fast repolarization afterspike, the duration of the refractory period, and voltage threshold accommodation, if present.

Actions of antipsychotic drugs on pancreatic β-cell function: contrasting effects of clozapine and haloperidol

The use of antipsychotic drugs is known to be associated with a number of adverse metabolic side effects, including diabetes mellitus. These side effects could be, at least in part, the result of impaired islet cell function, although the underlying mechanisms are unknown. We have studied the effects of the atypical antipsychotic clozapine and of the conventional drug haloperidol on electrical and secretory activity in rat pancreatic beta-cells. At a low glucose concentration (4mM), clozapine (5microM) had little or no effect on membrane potential. However, at a stimulatory glucose concentration (16mM), clozapine was found to hyperpolarize the cell membrane potential, resulting in a complete inhibition of electrical activity. In contrast, haloperidol (5microM) was found to cause a marked depolarization of the membrane potential in the presence of both low and high concentrations of glucose. Clozapine and haloperidol were found, respectively, to increase and decrease-cell input conductance, an index of K(+) permeability. Single channel recordings indicated that changes in K(ATP) channel activity contributed towards these effects. Neither clozapine nor haloperidol affected basal insulin release, although clozapine inhibited glucose-induced insulin release. It is concluded that clozapine and haloperidol exert contrasting actions on electrical activity in rat pancreatic -cells as a result of opposing effects on K(+) permeability. These findings may relate to the increased incidence of diabetes associated with clozapine treatment.

How Much Afterhyperpolarization Conductance Is Recruited by an Action Potential? A Dynamic-Clamp Study in Cat Lumbar Motoneurons

We accurately measured the conductance responsible for the afterhyperpolarization (medium AHP) that follows a single spike in spinal motoneurons of anesthetized cats. This was done by using the dynamic-clamp method. We injected an artificial current in the neurons that increased the AHP amplitude, and we made use of the fact that the intensity of the natural AHP current at the trough of the voltage trajectory was related linearly to the AHP amplitude. We determined at the same time the conductance and the reversal potential of the AHP current. This new method was validated by a simple theoretical model incorporating AHP and hyperpolarization-activated (Ih) currents and could be applied when the decay time constant of the AHP conductance was at least five times shorter than the estimated Ih activation time. This condition was fulfilled in 33 of 44 motoneurons. The AHP conductance varied from 0.3 to 1.4 microS in both slow- and fast-type motoneurons, which was approximately the same range as the input conductance of the entire population. However, AHP and input conductances were not correlated. The larger AHP in slow-type motoneurons was mainly attributable to their smaller input conductance compared with fast motoneurons. The likeness of the AHP conductance in both types of motoneurons is in sharp contrast to differences in AHP decay time and explains why slow- and fast-type motoneurons have similar gain.

Contributions of Intrinsic Membrane Dynamics to Fast Network Oscillations With Irregular Neuronal Discharges

During fast oscillations in the local field potential (40-100 Hz gamma, 100-200 Hz sharp-wave ripples) single cortical neurons typically fire irregularly at rates that are much lower than the oscillation frequency. Recent computational studies have provided a mathematical description of such fast oscillations, using the leaky integrate-and-fire (LIF) neuron model. Here, we extend this theoretical framework to populations of more realistic Hodgkin-Huxley-type conductance-based neurons. In a noisy network of GABAergic neurons that are connected randomly and sparsely by chemical synapses, coherent oscillations emerge with a frequency that depends sensitively on the single cell's membrane dynamics. The population frequency can be predicted analytically from the synaptic time constants and the preferred phase of discharge during the oscillatory cycle of a single cell subjected to noisy sinusoidal input. The latter depends significantly on the single cell's membrane properties and can be understood in the context of the simplified exponential integrate-and-fire (EIF) neuron. We find that 200-Hz oscillations can be generated, provided the effective input conductance of single cells is large, so that the single neuron's phase shift is sufficiently small. In a two-population network of excitatory pyramidal cells and inhibitory neurons, recurrent excitation can either decrease or increase the population rhythmic frequency, depending on whether in a neuron the excitatory synaptic current follows or precedes the inhibitory synaptic current in an oscillatory cycle. Detailed single-cell properties have a substantial impact on population oscillations, even though rhythmicity does not originate from pacemaker neurons and is an emergent network phenomenon.

Increased persistent Na+ current and its effect on excitability in motoneurones cultured from mutant SOD1 mice

Mutations in the enzyme superoxide dismutase 1 (SOD1) initiate a progressive motoneurone degeneration in amyotrophic lateral sclerosis (ALS). Transgenic mice overexpressing this mutation develop a similar progressive motoneurone degeneration. In spinal motoneurones cultured from presymptomatic mice expressing the glycine to alanine mutation at base pair 93 (G93A) SOD1 mutation, a marked increase in the persistent component of the Na(+) current was observed, without changes in passive properties. This increase only enhanced neuronal excitability in high input conductance cells, as low input conductance cells exhibited a compensatory outward shift in the current remaining after Na(+) blockade. High input conductance motoneurones tend to be large, so these results may explain the tendency of large motoneurones to degenerate first in ALS. Riluzole, at the therapeutic concentration used to treat ALS, decreased neuronal excitability and persistent Na(+) current in G93A motoneurones to levels observed in the control motoneurones. Aberrations in the intrinsic electrical properties may be among the first symptoms to emerge in SOD1-linked ALS.

Possible Effects of Depolarizing GABAA Conductance on the Neuronal Input–Output Relationship: A Modeling Study

Recent in vitro experiments revealed that the GABAA reversal potential is about 10 mV higher than the resting potential in mature mammalian neocortical pyramidal cells; thus GABAergic inputs could have facilitatory, rather than inhibitory, effects on action potential generation under certain conditions. However, how the relationship between excitatory input conductances and the output firing rate is modulated by such depolarizing GABAergic inputs under in vivo circumstances has not yet been understood. We examine herewith the input-output relationship in a simple conductance-based model of cortical neurons with the depolarized GABAA reversal potential, and show that a tonic depolarizing GABAergic conductance up to a certain amount does not change the relationship between a tonic glutamatergic driving conductance and the output firing rate, whereas a higher GABAergic conductance prevents spike generation. When the tonic glutamatergic and GABAergic conductances are replaced by in vivo-like highly fluctuating inputs, on the other hand, the effect of depolarizing GABAergic inputs on the input-output relationship critically depends on the degree of coincidence between glutamatergic input events and GABAergic ones. Although a wide range of depolarizing GABAergic inputs hardly changes the firing rate of a neuron driven by noncoincident glutamatergic inputs, a certain range of these inputs considerably decreases the firing rate if a large number of driving glutamatergic inputs are coincident with them. These results raise the possibility that the depolarized GABAA reversal potential is not a paradoxical mystery, but is instead a sophisticated device for discriminative firing rate modulation.

Systematic variation in effects of serotonin and norepinephrine on repetitive firing properties of ventral horn neurons

Spinal interneurons are essential integrators of descending and peripheral input that receive profuse monoaminergic influence from brainstem nuclei. In this study, the effects of the monoamines serotonin and norepinephrine on the intrinsic properties of ventral horn interneurons were investigated in a slice preparation of the lumbar cord of 7–19 day old rats. Three cell groups with distinct firing patterns in response to steps of injected current were observed and classified as repetitive-firing, initial-burst or single-spiking. Input conductance tended to be largest in single-spiking cells whereas repetitive-firing cells showed the greatest tendency for spontaneous firing and had the fastest rate of rise for the action potential. Rhythmic firing behaviors were defined by the frequency-current relation evoked by linearly increasing current ramps. The monoaminergic modulation of firing patterns and frequency-current relations was primarily studied in repetitive-firing cells. The frequency-current threshold current was decreased in cells with high pre-drug values and increased in cells with low pre-drug values. Therefore, monoamine administration decreased the input-output heterogeneity of the repetitive-firing cells by compressing the range of frequency-current threshold currents. This action of monoamines may have a key role in the suppression of sensory-evoked reflexes and the production of coordinated movement.

Angiotensin II Excites Paraventricular Nucleus Neurons That Innervate the Rostral Ventrolateral Medulla: An In Vitro Patch-Clamp Study in Brain Slices

Neurons of the hypothalamic paraventricular nucleus (PVN) are key controllers of sympathetic nerve activity and receive input from angiotensin II (ANG II)-containing neurons in the forebrain. This study determined the effect of ANG II on PVN neurons that innervate in the rostral ventrolateral medulla (RVLM)-a brain stem site critical for maintaining sympathetic outflow and arterial pressure. Using an in vitro brain slice preparation, whole cell patch-clamp recordings were made from PVN neurons retrogradely labeled from the ipsilateral RVLM of rats. Of 71 neurons tested, 62 (87%) responded to ANG II. In current-clamp mode, bath-applied ANG II (2 muM) significantly (P < 0.05) depolarized membrane potential from -58.5 +/- 2.5 to -54.5 +/- 2.0 mV and increased the frequency of action potential discharge from 0.7 +/- 0.3 to 2.8 +/- 0.8 Hz (n = 4). Local application of ANG II by low-pressure ejection from a glass pipette (2 pmol, 0.4 nl, 5 s) also elicited rapid and reproducible excitation in 17 of 20 cells. In this group, membrane potential depolarization averaged 21.5 +/- 4.1 mV, and spike activity increased from 0.7 +/- 0.4 to 21.3 +/- 3.3 Hz. In voltage-clamp mode, 41 of 47 neurons responded to pressure-ejected ANG II with a dose-dependent inward current that averaged -54.7 +/- 3.9 pA at a maximally effective dose of 2.0 pmol. Blockade of ANG II AT1 receptors significantly reduced discharge (P < 0.001, n = 5), depolarization (P < 0.05, n = 3), and inward current (P < 0.01, n = 11) responses to locally applied ANG II. In six of six cells tested, membrane input conductance increased (P < 0.001) during local application of ANG II (2 pmol), suggesting influx of cations. The ANG II current reversed polarity at +2.2 +/- 2.2 mV (n = 9) and was blocked (P < 0.01) by bath perfusion with gadolinium (Gd(3+), 100 muM, n = 8), suggesting that ANG II activates membrane channels that are nonselectively permeable to cations. These findings indicate that ANG II excites PVN neurons that innervate the ipsilateral RVLM by a mechanism that depends on activation of AT1 receptors and gating of one or more classes of ion channels that result in a mixed cation current.

How shunting inhibition affects the discharge of lumbar motoneurones: a dynamic clamp study in anaesthetized cats.

In the present work, dynamic clamp was used to inject a current that mimicked tonic synaptic activity in the soma of cat lumbar motoneurones with a microelectrode. The reversal potential of this current could be set at the resting potential so as to prevent membrane depolarization or hyperpolarization. The only effect of the dynamic clamp was then to elicit a constant and calibrated increase of the motoneurone input conductance. The effect of the resulting shunt was investigated on repetitive discharges elicited by current pulses. Shunting inhibition reduced very substantially the firing frequency in the primary range without changing the slope of the current-frequency curves. The shift of the I-f curve was proportional to the conductance increase imposed by the dynamic clamp and depended on an intrinsic property of the motoneurone that we called the shunt potential. The shunt potential ranged between 11 and 37 mV above the resting potential, indicating that the sensitivity of motoneurones to shunting inhibition was quite variable. The shunt potential was always near or above the action potential voltage threshold. A theoretical model allowed us to interpret these experimental results. The shunt potential was shown to be a weighted time average of membrane voltage. The weighting factor is the phase response function of the neurone that peaks at the end of the interspike interval. The shunt potential indicates whether mixed synaptic inputs have an excitatory or inhibitory effect on the ongoing discharge of the motoneurone.

Inhibition of glucose-induced electrical activity in rat pancreatic β-cells by DCPIB, a selective inhibitor of volume-sensitive anion currents

We have investigated the effects of the ethacrynic acid derivative 4-(2-butyl-6,7-dichloro-2-cyclopentyl-indan-1-on-5-yl) oxobutyric acid (DCPIB), an inhibitor of the volume-sensitive anion channel (VSAC), on electrical activity and insulin secretion in rat pancreatic β-cells. DCPIB inhibited whole-cell VSAC currents in β-cells with IC 50 values of 2.2 and 1.7 μM for inhibition of outward and inward currents, respectively. DCPIB also inhibited the VSAC at the single channel level in cells activated by glucose. In intact cells, DCPIB caused a net increase in β-cell input conductance and evoked an outward current that was sensitive to inhibition by tolbutamide, suggesting K ATP channel activation. However, no K ATP channel activation was evident under conventional whole-cell conditions, suggesting that the drug might activate the channel in intact cells via an indirect mechanism, possibly involving nutrient metabolism. DCPIB suppressed glucose-induced electrical activity in β-cells, hyperpolarised the cell membrane potential at a substimulatory glucose concentration and prevented depolarisation when the glucose concentration was raised to stimulatory levels. The suppression of electrical activity by DCPIB was associated with a marked inhibition of glucose-stimulated insulin release from intact islets. It is concluded that DCPIB inhibits electrical and secretory activity in the β-cell as a combined result of a reciprocal inhibition of VSAC and activation of K ATP channel activities, thus producing a marked hyperpolarisation of the β-cell membrane potential.

Neuronal integration of synaptic input in the fluctuation-driven regime.

During sensory stimulation, visual cortical neurons undergo massive synaptic bombardment. This increases their input conductance, and action potentials mainly result from membrane potential fluctuations. To understand the response properties of neurons operating in this regime, we studied a model neuron with synaptic inputs represented by transient membrane conductance changes. We show that with a simultaneous increase of excitation and inhibition, the firing rate first increases, reaches a maximum, and then decreases at higher input rates. Comodulation of excitation and inhibition, therefore, does not provide a straightforward way of controlling the neuronal firing rate, in contrast to coding mechanisms postulated previously. The synaptically induced conductance increase plays a key role in this effect: it decreases firing rate by shunting membrane potential fluctuations, and increases it by reducing the membrane time constant, allowing for faster membrane potential transients. These findings do not depend on details of the model and, hence, are relevant to cells of other cortical areas as well.

Long-chain CoA esters activate human pancreatic beta-cell KATP channels: potential role in Type 2 diabetes.

The ATP-regulated potassium (KATP) channel in the pancreatic beta cell couples the metabolic state to electrical activity. The primary regulator of the KATP channel is generally accepted to be changes in ATP/ADP ratio, where ATP inhibits and ADP activates channel activity. Recently, we showed that long-chain CoA (LC-CoA) esters form a new class of potent KATP channel activators in rodents, as studied in inside-out patches. In this study we have investigated the effects of LC-CoA esters in human pancreatic beta cells using the inside-out and whole-cell configurations of the patch clamp technique. Human KATP channels were potently activated by acyl-CoA esters with a chain length exceeding 12 carbons. Activation by LC-CoA esters did not require the presence of Mg2+ or adenine nucleotides. A detailed characterization of the concentration-dependent relationship showed an EC50 of 0.7+/-0.1 micromol/l. Furthermore, in the presence of an ATP/ADP ratio of 10 (1.1 mmol/l total adenine nucleotides), whole-cell KATP channel currents increased approximately six-fold following addition of 1 micro mol/l LC-CoA ester. The presence of 1 micro mol/l LC-CoA in the recording pipette solution increased beta-cell input conductance, from 0.5+/-0.2 nS to 2.5+/-1.3 nS. Taken together, these results show that LC-CoA esters are potent activators of the KATP channel in human pancreatic beta cells. The fact that LC-CoA esters also stimulate KATP channel activity recorded in the whole-cell configuration, points to the ability of these compounds to have an important modulatory role of human beta-cell electrical activity under both physiological and pathophysiological conditions.

Quantification of the factors that influence discharge correlation in model motor neurons.

The purpose of this study was to quantify the influence of intrinsic properties, active dendritic conductances, and background excitation and inhibition on measures of discharge correlation in the time and frequency domains with known levels and patterns of common synaptic input. The study involved a computer simulation of a population of neurons with a range of input resistances (0.54-3.7 MOmega) and surface areas (407,000-712,000 microm(2)). The neurons were simulated with no, moderate, or high levels of active dendritic conductances and were activated with either excitatory input only or excitatory and inhibitory inputs. The patterns of common input, either branched common input or common modulation, were tested with 0, 30, 60, and 90% common input. The results confirm previous findings of an exponential relation between the level of common input and indexes of synchronization; only when the common input comprised >/=60% of the total excitatory input was there a significant effect on discharge correlation. Synchronization was greatest in models that had passive dendrites. Active dendritic conductances caused the discharge rate of the neuron to saturate and decreased motor-unit synchronization. However, the addition of 10% background inhibitory input increased synchronization in these models. In contrast, common rhythmic modulation of inputs at 24 Hz usually decreased synchronization. Significant coherence at the modulated frequency occurred in the commonly modulated neurons when >/=60% of the inputs were modulated. Furthermore, active dendritic conductances decreased coherence. Branched common input caused high levels of coherence across a broad spectrum and when combined with active dendritic conductances caused significant frequency peaks in the 30- to 50-Hz band. In conclusion, the level of inhibitory input and active dendritic conductances interact with the amount of common input to determine time- and frequency-domain discharge correlation.

Pentobarbital modulates intrinsic and gaba-receptor conductances in thalamocortical inhibition

We investigated interactions of an anesthetic barbiturate, pentobarbital, with non-ligand gated channels and identified inhibitory synaptic transmission in thalamic neurons. Using whole cell voltage-clamp, current-clamp and single channel recording techniques in rat ventrobasal neurons of slices and dispersed preparations, we determined the mechanisms of pentobarbital actions on ionic currents and inhibitory postsynaptic currents (IPSCs), mediated by aminobutyric acid (GABA). We investigated pentobarbital effects on intrinsic currents using hyperpolarizing voltage commands from rest and tetrodotoxin blockade of action potentials. At concentrations near 8 μM, pentobarbital increased input conductance and induced net outward current, I PB , at potentials near action potential threshold. The reversal potential of I PB was −75 mV, implicating K + and other ions. Cs + (3 mM) which inhibits both K + currents and inward rectifier (I h ), completely blocked I PB , whereas the selective I h blocker, ZD-7288 (25 μM), or Ba 2+ (2 mM) which suppresses only K + currents, reduced I PB . Pentobarbital inhibited the I h , consistent with a ZD-7288-induced shift in reversal potential for I PB toward K + equilibrium potential. Pentobarbital increased the inward K + rectifier, I Kir , and leak current, I leak . We determined the susceptibility of IPSCs, evoked by reticular stimulation, to antagonism by bicuculline, picrotoxinin and 2-hydroxysaclofen and identified their receptor subclass components. At EC 50 =53 μM, pentobarbital increased the duration of IPSCs. The prolonged IPSC duration during pentobarbital was attributable to enhanced open probability of GABA A channels, because combined with GABA, pentobarbital application increased mean channel open time without affecting channel conductance. At concentrations up to 100 μM, pentobarbital did not directly activate GABA A receptors. The concentration-response relationships for pentobarbital effects on the intrinsic currents and IPSCs overlapped, implying multiple sites of action and possible redundancy in anesthetic mechanisms. This is the first study to show that an i.v. anesthetic modulates the intrinsic currents, I h , I Kir , and I leak , as well as IPSC time course in the same neurons. These effects likely underlie inhibition in thalamocortical neurons during pentobarbital anesthesia.

Pentobarbital modulates intrinsic and GABA-receptor conductances in thalamocortical inhibition

We investigated interactions of an anesthetic barbiturate, pentobarbital, with non-ligand gated channels and identified inhibitory synaptic transmission in thalamic neurons. Using whole cell voltage-clamp, current-clamp and single channel recording techniques in rat ventrobasal neurons of slices and dispersed preparations, we determined the mechanisms of pentobarbital actions on ionic currents and inhibitory postsynaptic currents (IPSCs), mediated by aminobutyric acid (GABA). We investigated pentobarbital effects on intrinsic currents using hyperpolarizing voltage commands from rest and tetrodotoxin blockade of action potentials. At concentrations near 8 μM, pentobarbital increased input conductance and induced net outward current, I PB, at potentials near action potential threshold. The reversal potential of I PB was −75 mV, implicating K + and other ions. Cs + (3 mM) which inhibits both K + currents and inward rectifier (I h), completely blocked I PB, whereas the selective I h blocker, ZD-7288 (25 μM), or Ba 2+ (2 mM) which suppresses only K + currents, reduced I PB. Pentobarbital inhibited the I h, consistent with a ZD-7288-induced shift in reversal potential for I PB toward K + equilibrium potential. Pentobarbital increased the inward K + rectifier, I Kir, and leak current, I leak. We determined the susceptibility of IPSCs, evoked by reticular stimulation, to antagonism by bicuculline, picrotoxinin and 2-hydroxysaclofen and identified their receptor subclass components. At EC 50=53 μM, pentobarbital increased the duration of IPSCs. The prolonged IPSC duration during pentobarbital was attributable to enhanced open probability of GABA A channels, because combined with GABA, pentobarbital application increased mean channel open time without affecting channel conductance. At concentrations up to 100 μM, pentobarbital did not directly activate GABA A receptors. The concentration-response relationships for pentobarbital effects on the intrinsic currents and IPSCs overlapped, implying multiple sites of action and possible redundancy in anesthetic mechanisms. This is the first study to show that an i.v. anesthetic modulates the intrinsic currents, I h, I Kir, and I leak, as well as IPSC time course in the same neurons. These effects likely underlie inhibition in thalamocortical neurons during pentobarbital anesthesia.