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W J Wadman

Publications and source records attributed to W J Wadman.

At least 37 records · Page 2Linked to original sources
PubMed1998

Sodium currents in isolated rat CA1 neurons after kindling epileptogenesis.

Cellular excitability of CA1 neurons from a kindled focus in the rat hippocampus is persistently increased. The changes in the underlying voltage-dependent sodium current were characterized under whole-cell voltage-clamp conditions. We compared sodium currents in acutely isolated CA1 neurons from kindled rats with those in matched controls, one day and five weeks after cessation of kindling stimulations. The sodium current in CA1 neurons was tetrodotoxin sensitive and inactivated completely with two time-constants. In 97 cells from control rats, the current evoked at -20 mV consisted of a fast-inactivating component of 3.8 +/- 0.2 nA which decayed with a time-constant of 1.0 +/- 0.1 ms, and a slow-inactivating component of 1.2 +/- 0.1 nA with a time-constant of 3.6 +/- 0.1 ms. The potential of half-maximal inactivation was -72.2 +/- 1.0 mV for the fast-inactivating component and -63.2 +/- 1.0 mV for the slow-inactivating component. The time-constant of recovery at -80 mV was 14.1 +/- 0.4 ms for the fast-inactivating component and 9.3 +/- 0.4 ms for the slow-inactivating component. One day after kindling, the voltage dependence of inactivation of the slow-inactivating and the fast-inactivating component was shifted in the depolarizing direction (3.2 +/- 1.3 and 3.0 +/- 1.3 mV, respectively). The voltage dependence of recovery from inactivation was shifted in the same direction. Five weeks after kindling, the shift in voltage dependence of inactivation was (3.3 +/- 1.2 and 2.9 +/- 1.2 mV, respectively) and was accompanied by a 20% increase in sodium current amplitude. The voltage-dependent activation was not different after kindling. The changes in sodium current inactivation will increase the number of channels available for activation and may enhance the maximum firing rate. This implies that the changes in sodium current inactivation will contribute to the enhanced excitability of pyramidal neurons observed after kindling.

Animals↗
PubMed1998

Effect of valproic acid on sodium currents in cortical neurons from patients with pharmaco-resistant temporal lobe epilepsy.

In a selected group of temporal lobe epilepsy patients with seizures refractory to pharmacological treatment, pharmacological seizure control can be attained by surgical resection of the epileptic zone. We investigated to what extent pharmaco-resistance is reflected in a reduced response at the cellular level, in neurons acutely isolated from the temporal cortex resected in 20 patients. We studied the effect of valproic acid (VPA) on the transient sodium current, measured under whole-cell voltage-clamp conditions. We compared neurons from patients with temporal lobe sclerosis (S) with neurons from patients without hippocampal sclerosis (nS) and compared hippocampal CA1 neurons (CA) with neocortical neurons (NC). We could not detect differences in the voltage dependence and kinetics of sodium current activation and inactivation in any of the group comparisons. VPA shifted the voltage dependence of steady-state inactivation (expressed as V(h,i) in a Boltzmann fit) to more hyperpolarized levels. The shift induced by 2 mM VPA was -5.1 +/- 0.7 mV in CA-S (n = 13), -5.1 +/- 0.7 mV in CA-nS (n = 25), -4.3 +/- 0.5 mV in NC-S (n = 17) and -4.9 +/- 0.5 mV in NC-nS (n = 16) The relation between concentration and voltage shift had an EC50 of 1.4 +/- 0.2 mM VPA (n = 16) and a maximal shift of 9.6 +/- 0.9 mV. We conclude that pharmaco-resistance in these patients is not associated with a changed modulation of the sodium current by VPA. Results are discussed in the light of a reduced sodium current modulation by carbamazepine in CA1 neurons of patients with hippocampal sclerosis and of similar observations in the kindling model of epileptogenesis.

Anticonvulsants↗
PubMed1997

Development of HVA and LVA calcium currents in pyramidal CA1 neurons in the hippocampus of the rat.

High voltage activated (HVA) and low voltage activated (LVA) calcium currents were recorded in acutely dissociated CA1 hippocampal pyramidal neurons of the rat during the first three postnatal weeks as well as in adults. Measured in whole cell voltage clamp the amplitude of the HVA calcium current increased steadily and reached adult values after 20 postnatal days (P20). Using the perforated patch configuration with amphotericin B the amplitude of the HVA component was more than five times smaller, but the time course of development was the same. The LVA component also increased with age but reached adult values already around P13. The amplitude and developmental pattern of this component were not different when measured with the perforated patch technique. The results indicate a different role for intracellular modulators on these calcium currents, but exclude them as important factors in the developmental pattern. The fast development of the LVA component could lead to calcium dependent action potentials (and calcium spikes) in immature cells. The complex developmental pattern of the relative amplitude of the two currents will either lead to specific variations in the intracellular calcium homeostasis or will have to be accompanied by an adequate developmental pattern of buffering and extrusion mechanisms.

Amphotericin B↗
PubMed1997

High intracellular calcium levels during and after electrical discharges in molluscan peptidergic neurons.

Intracellular calcium levels ([Ca2+]i) during and following electrical activity of the neuroendocrine caudodorsal cells of the pond snail (Lymnaea stagnalis) were measured in situ and in dissociated cells by combining electrical recordings and Fura-2 measurements. Caudodorsal cells are typical neuroendocrine cells that control egg laying via the release of a set of peptides during a stereotyped discharge of action potentials. Single action potentials or short trains of spikes in dissociated caudodorsal cells induced only small but consistent increases in [Ca2+]i. With longer or repeated spike trains, larger [Ca2+]i transients were measured, indicating accumulation of calcium. The calcium channel blocker Ni2+ suppressed the calcium elevation, suggesting that calcium influx occurred through voltage-activated calcium channels. Calcium levels in caudodorsal cells in situ were measured before, during and after the stereotyped firing pattern, a approximately 35-min discharge of regular spiking. Basal calcium levels in caudodorsal cells in situ were about 125 nM. During the initial phase of the discharge, the intracellular calcium level increased to approximately 250 nM. Maximal calcium levels (300-600 nM) were only reached at the final phase of the discharge or several minutes after the cessation of firing. Calcium levels remained elevated for up to 1 h after the end of the discharge. During this time, caudodorsal cells were characterized by very low excitability. We suggest that the prolonged, elevated level of calcium following the discharge need not be directly dependent on action potentials. The long-lasting [Ca2+]i elevation may cause the release of neuropeptides to outlast the duration of electrical activity, thus uncoupling release from spiking. In addition, it may cause reduced excitability of neuroendocrine cells following a period of spiking, thereby inducing a refractory period.

Action Potentials↗
PubMed1997

High intracellular calcium levels during and after electrical discharges in molluscan peptidergic neurons.

Intracellular calcium levels ([Ca2+]i) during and following electrical activity of the neuroendocrine caudodorsal cells of the pond snail (Lymnaea stagnalis) were measured in situ and is dissociated cells by combining electrical recordings and Fura-2 measurements. Caudodorsal cells are typical neuroendocrine cells that control egg laying via the release of a set of peptides during a stereotyped discharge of action potentials. Single action potentials or short trains of spikes in dissociated caudodorsal cells induced only small but consistent increases in [Ca2+]i. With longer or repeated spike trains, larger [Ca2+]i transients were measured, indicating accumulation of calcium. The calcium channel blocker Ni2+ suppressed the calcium elevation, suggesting that calcium influx occurred through voltage-activated calcium channels. Calcium levels in caudodorsal cells in situ were measured before, during and after the stereotyped firing pattern, a approximately 35-min discharge of regular spiking. Basal calcium levels in caudodorsal cells in situ were about 125 nM. During the initial phase of the discharge, the intracellular calcium level increased to approximately 250 nM. Maximal calcium levels (300-600 nM) were only reached at the final phase of the discharge or several minutes after the cessation of firing. Calcium levels remained elevated for up to 1 h after the end of the discharge. During this time, caudodorsal cells were characterized by very low excitability. We suggest that the prolonged, elevated level of calcium following the discharge need not be directly dependent on action potentials. The long-lasting [Ca2+]i elevation may cause the release of neuropeptides to outlast the duration of electrical activity, thus uncoupling release from spiking. In addition, it may cause reduced excitability of neuroendocrine cells following a period of spiking, thereby inducing a refractory period.

Action Potentials↗
PubMed1996

Polyunsaturated fatty acids modulate sodium and calcium currents in CA1 neurons.

Recent evidence indicates that long-chain polyunsaturated fatty acids (PUFAs) can prevent cardiac arrhythmias by a reduction of cardiomyocyte excitability. This was shown to be due to a modulation of the voltage-dependent inactivation of both sodium (INa) and calcium (ICa) currents. To establish whether PUFAs also regulate neuronal excitability, the effects of PUFAs on INa and ICa were assessed in CA1 neurons freshly isolated from the rat hippocampus. Extracellular application of PUFAs produced a concentration-dependent shift of the voltage dependence of inactivation of both INa and ICa to more hyperpolarized potentials. Consequently, they accelerated the inactivation and retarded the recovery from inactivation. The EC50 for the shift of the INa steady-state inactivation curve was 2.1 +/- 0.4 microM for docosahexaenoic acid (DHA) and 4 +/- 0.4 microM for eicosapentaenoic acid (EPA). The EC50 for the shift on the ICa inactivation curve was 2.1 +/- 0.4 for DHA and > 15 microM for EPA. Additionally, DHA and EPA suppressed both INa and ICa amplitude at concentrations > 10 microM. PUFAs did not affect the voltage dependence of activation. The monounsaturated oleic acid and the saturated palmitic acid were virtually ineffective. The combined effects of the PUFAs on INa and ICa may reduce neuronal excitability and may exert anticonvulsive effects in vivo.

Animals↗
PubMed1996

Calcium currents in pyramidal CA1 neurons in vitro after kindling epileptogenesis in the hippocampus of the rat.

Calcium is an important second messenger which plays a role in the regulation of neuronal excitability and in many forms of synaptic plasticity. In kindling epileptogenesis, a model of focal epilepsy, calcium plays an important role. The in situ patch-clamp technique was used to record calcium currents in slices obtained from kindled rats and controls. We found that low-voltage-activated calcium currents, probably of dendritic origin, were larger after kindling (80%). The transient high-voltage-activated calcium currents were also enhanced after kindling (50% higher). The increase of the current is accompanied by a decrease in the time constant of inactivation. The change was still present six weeks after the kindling stimulations were stopped. These data demonstrate that low-voltage-activated calcium currents are involved in epileptogenesis. Their enhancement in the dendrites will boost synaptic depolarization and result in enhanced calcium influx, which is critically dependent on the specific activation pattern.

Action Potentials↗
PubMed1995

Effects of strongly anisosmotic and NaCl deficient solutions on muscimol- and glutamate evoked whole-cell currents in freshly dissociated hippocampal neurons.

Sudden exposure of dissociated hippocampal neurons to strongly hypo- or hyperosmotic solutions suppresses voltage gated Na+, K+ and Ca2+ currents. We investigated whether ligand gated ion currents were similarly shut down by exposure to anisosmotic solutions. The effect of hypo-osmotic, NaCl deficient (mannitol-substituted), or hyper-osmotic test solutions delivered from a flow pipette was tested on voltage gated Ca2+ currents and on currents and conductance changes evoked by brief administration of either the GABAA-agonist muscimol or glutamate. Hyper-osmotic solution caused cells to shrink, but cell membrane capacitance did not change. Muscimol-induced conductance increases were depressed by hypo-osmotic and by NaCl deficient solutions and often by hyper-osmotic solution. Voltage gated Ca2+ currents were depressed by anisosmotic, but not by NaCl deficient isosmotic solution. NMDA- and non-NMDA evoked conductance increases were depressed by hyperosmotic solution; hypo-osmotic and NaCl deficient solutions were not tested on glutamate induced currents. Ligand gated currents are suppressed by anisosmotic solutions more slowly than are voltage gated channels. The changes caused by anisosmotic and NaCl deficient solutions were much greater then expected from calculated electrochemical effects and are probably the result of change in receptor controlled channels.

Animals↗
PubMed1995

Potassium currents in isolated CA1 neurons of the rat after kindling epileptogenesis.

Daily tetanic stimulation of the Schaffer collaterals generates an epileptogenic focus in area CA1 of the rat hippocampus, ultimately leading to generalized tonic-clonic convulsions (kindling). Potassium currents were measured under voltage-clamp conditions in pyramidal neurons, acutely dissociated from the focus of fully kindled rats, one day and six weeks after the last generalized seizure. Their amplitude, kinetics, voltage dependence and calcium dependence were compared with controls. With Ca2+ influx blocked by 0.5 mM Ni2+, the sustained current (delayed rectifier) and the transient current (A-current) were not different after kindling. Calcium influx evoked an additional fast transient current component. This transient calcium-dependent current component was increased by 154%, but only immediately after the seizure. A second, slow calcium-dependent potassium current component was dependent on the intracellular calcium level, set by the pipette as well as on calcium influx. The peak amplitude of this slow calcium-dependent current was under optimal calcium conditions not different after kindling, but we found indications that either calcium homeostasis or the calcium sensitivity of the potassium channels was affected by the kindling process. In contrast to the previously described enhancement of calcium current, kindling epileptogenesis did not change the total potassium current amplitude. The minor changes that were observed can be related either to changes in calcium current or to changes in intracellular calcium homeostasis.

Animals↗
PubMed1995

Development of voltage-activated potassium currents in cultured cerebellar granule neurons under different growth conditions.

1. The functional expression of two potassium currents in cultured cerebellar granule cells was investigated with the whole cell patch-clamp technique in relation to development and growth condition. Cells were grown in medium containing different concentrations of potassium: 25 mM (K25) and 40 mM (K40), together referred to as "high K+"; 10 mM (K10) or "low K+"; and K10 with 100 microM N-methyl-D-aspartate (KNMDA). All conditions are known to influence maturation and survival of granule cells in culture. 2. At 2 days in vitro (DIV) the membrane capacitance, taken as index of membrane surface area, was the same for cells grown in each growth condition. At 7-9 DIV it had increased in each condition, but to a substantially larger extent in cells grown in KNMDA, K25, and K40 than in cells grown in K10. During development the input resistance only decreased in cells grown in KNMDA and high K+. 3. A delayed potassium current (IK) and a fast transient potassium current (IA) could both be recorded at 2 DIV in each growth condition, although a few neurons only expressed the IK. The IK was partially suppressed by tetraethylammonium (5 mM), whereas IA was predominantly sensitive to 4-aminopyridine (5 mM). 4. Normalized for cell capacitance, the specific IA conductance hardly changed during development in cells grown in high K+ and KNMDA. Cells in K10, however, displayed an IA with totally different properties in 23 of 24 cells; the specific IA conductance in these cells was considerably smaller at 7-9 DIV, suggesting a deletion of these channels during development.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Philanthotoxin inhibits Ca2+ currents in rat hippocampal CA1 neurons.

The wasp venom philanthotoxin-4.3.3 (PhTX-4.3.3) is an antagonist of glutamate transmission in the insect as well as in the mammalian brain. It was recently shown that PhTX-4.3.3 inhibits the N-methyl-D-aspartate (NMDA) transmission in rat hippocampus. In this study we show that dideaza-philanthotoxin-12 (dideaza-PhTX-12), an analogue of PhTX-4.3.3, is a potent antagonist of voltage-dependent Ca2+ currents in rat hippocampal CA1 neurons. At a concentration of 10 microM it reduces the Ca2+ current to 40%. Two voltage-dependent potassium currents, the A current and the delayed rectifier, were hardly affected by dideaza-PhTX-12, indicating selectivity of the drug for Ca2+ currents. As a consequence the philanthotoxins will inhibit the calcium influx via voltage dependent as well as NMDA mediaded calcium channels and thus reduce excitability in the hippocampus.

Animals↗
PubMed1994

Corticosteroid receptor-dependent modulation of calcium currents in rat hippocampal CA1 neurons.

Pyramidal CA1 neurons in the rat hippocampus contain mineralocorticoid (MRs) and glucocorticoid receptors (GRs) for corticosterone, which, in activated form, act as transcription factors of the genome. The relative MR and GR occupation changes throughout the day, with predominant MR occupation under rest in the morning and additional GR occupation in the evening and after stress. We examined the effect of MR and GR activation on Ca currents in hippocampal slices from adrenalectomized (ADX) rats under whole-cell voltage-clamp conditions. In slices from ADX rats, where MRs and GRs are unoccupied, Ca currents (particularly in the low-voltage range) were larger than in neurons from the sham-operated controls; these effects became apparent with a delay of > or = 3 days after ADX. Selective occupation of MRs in tissue from ADX rats greatly (by 70%) and persistently (up to 3 h) reduced transient but also sustained Ca conductances. Voltage dependency and kinetic properties of the currents were not affected. Occupation of GRs as well as MRs by corticosterone (30 nM) resulted in relatively large Ca currents, comparable to those recorded in tissue from mildly stressed sham-operated control animals. Interestingly, exclusive occupation of GRs with 30 nM RU 28362 was not sufficient to induce large Ca currents. The data suggest that the changes in MR and GR occupation throughout the day, related to circadian and stress-induced corticosterone release, are linked to marked alterations in Ca currents, with small Ca currents in the morning and large currents in the evening or after stress.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Cortex Hormones↗
PubMed1994

Kindling-induced long-lasting enhancement of calcium current in hippocampal CA1 area of the rat: relation to calcium-dependent inactivation.

Daily tetanization of the Schaffer collaterals (kindling) in the rat hippocampus induces a persistent epileptogenic focus in area CA1. Neurons were enzymatically isolated from the focal region one day or six weeks after seven class V generalized seizures had been evoked. Calcium currents were measured under voltage-clamp conditions in the whole-cell patch configuration. One day after kindling, as well as six weeks later, the amplitudes of a slow-inactivating (tau = 90 ms) and a non-inactivating calcium current component were, in comparison to controls, enhanced by 30 and 40%, respectively. This enhancement was therefore related to the kindled state of enhanced excitability. The enhancement of the calcium current was independent of the steady-state intracellular calcium concentration. Fast calcium-dependent inactivation was provoked with double-pulse protocols that conditioned the neuron with a defined calcium-influx in the first pulse. Despite the larger calcium current during the conditioning pulse, the relative calcium-dependent inactivation of the sustained current component was reduced in neurons from the kindled focus. Repetitive depolarizations, once every second, evoked a cumulative calcium-dependent inactivation. Nothwithstanding the larger calcium current, kindling also persistently reduced this slow inactivation of both transient and sustained high threshold calcium current. The reduction in calcium-dependent inactivation cannot be responsible for the increased current, but can certainly enhance the calcium influx during prolonged activation or seizures. The changes can be explained by assuming that additional calcium channels are recruited at a location that prevents calcium-dependent inactivation.

Animals↗
PubMed1994

Dynamics of local neuronal networks: control parameters and state bifurcations in epileptogenesis.

The aim of this overview is to present evidence that local neuronal networks (LNNs) are functionally organized in such a way that they behave as dynamic non-linear systems that can exhibit multiple types of attractor and can present bifurcations between different attractors, depending on control parameters. To begin with, some of the theoretical concepts of non-linear dynamics and chaos are briefly presented. As a case study, we described the CA1 area of the hippocampus and the changes that the corresponding LNNs undergo during kindling epileptogenesis. During epileptic seizures, evidence exists for the presence of low-dimensional chaos, since the correlation dimension estimated from the corresponding EEG signals decreases dramatically from a large value, characteristic of the resting state, to a low value typical of deterministic chaos. We propose that, among other things, an important control parameter of the dynamics of this brain area is the balance between excitatory (E) and inhibitory (I) processes. We assume that this balance can be experimentally estimated by using a paired-pulse paradigm. Accordingly, we demonstrate that the paired-pulse response changes during kindling epileptogenesis in the sense that the E/I ratio increases in the course of the establishment of a kindled epileptogenic focus. This change in E/I leads to a shift in the operating point of the LNN moving it close to a bifurcation where a rapid state change takes place. In this way, the LNN dynamics can change more readily to the basin of attraction of a chaotic attractor than under normal conditions. This is in essence what makes the behavior of the LNN more sensitive to tetanus, and predicts the facilitated occurrence of epileptic seizures during kindling.

Animals↗
PubMed1993

Channel shutdown: a response of hippocampal neurons to adverse environments.

Stretch-activated ion channels have been discovered in the membrane of many types of cells, but their presence in neurons is uncertain. We used freshly dissociated rat hippocampal neurons to study the effect of hypotonic swelling but, surprisingly, the isolated neurons did not swell. Voltage-dependent whole-cell membrane currents mediated by K+, Na+ and Ca2+ were rapidly and reversibly suppressed during sudden exposure to strongly hypo-osmotic, hyper-osmotic or glucose deficient solutions. The amplitudes of the sustained components of K+ and Ca2+ currents were more depressed than transient currents, but the rate of decay of transient K+ current greatly accelerated. The voltage dependence of activation and of steady state inactivation of residual K+ and Ca2+ currents were not shifted. The current holding membrane potential at -70 mV and therefore the conductance at that voltage were unchanged or somewhat decreased. Capacitive (charging) membrane current was not affected. Changes in tail current suggested moderate loss of cytosolic K+ in some but not in all cells. We conclude that channel shutdown is a uniform response of neuron somata and proximal dendrites to various adverse environments. Hypothetically we propose that swelling was prevented in anisosmotic conditions because membrane water permeability decreased.

Animals↗
PubMed1993

Low-threshold calcium current in dendrites of the adult rat hippocampus.

The patch-clamp technique was employed in hippocampal slices to examine the characteristics of a low-threshold Ca current in adult CA1 pyramidal neurons. We found that adult CA1 pyramidal neurons possess a distinct transient low-threshold Ca current, located predominantly in the distal dendrites. Surgical cuts that separated the dendrites from the soma and left a dendritic length of 150 microns, completely abolished the low-threshold Ca current while a transient K current persisted even in cells with dendrites as short as 50 microns. The transient low-threshold Ca current in dendrites may represent a voltage-dependent Ca entry pathway which contributes to the regulation of cellular excitability, plasticity and pathology.

Animals↗
PubMed1993

Bicuculline increases the intracellular calcium response of CA1 hippocampal neurons to synaptic stimulation.

Intracellular calcium levels were measured by ratio imaging of Fura-2-injected CA1 pyramidal neurons during repetitive electrical stimulation of the Schaffer collaterals in rat hippocampal slices. Baseline intracellular calcium levels of 102 nM increased to 190 nM during a 3-s stimulus train of 5 Hz. Bicuculline (20 microM) significantly enhanced this stimulus-dependent rise in intracellular calcium, while the baseline calcium levels remained unchanged. Concomitantly performed extra- and intracellular electrophysiological recordings indicate that the increased calcium response in the presence of bicuculline is linked to a prolongation of the excitatory postsynaptic potential and the induction of multiple action potentials. The bicuculline-induced increased calcium response could have long-term implications for cell function and eventually lead to cell degeneration.

Action Potentials↗