Minimal alterations in T-type calcium channel gating markedly modify physiological firing dynamics.

T-type calcium channel isoforms expressed in heterologous systems demonstrate marked differences in the biophysical properties of the resulting calcium currents. Such heterogeneity in gating behaviour not only reflects structural differences but is also observed following the regulation of channel activity by a number of ligands. However, the physiological impact of these differences in gating parameters of the T channels has never been evaluated in situ where the unique interplay between T-type calcium and other intrinsic currents is conserved, and T channel activation can be triggered by synaptic stimulation. Here, using the dynamic clamp technique, artificial T conductances were re-incorporated in thalamic neurons devoid of endogenous T currents to dissect the physiological role of the T current gating diversity on neuronal excitability. We demonstrate that the specific kinetics of the T currents in thalamocortical and nucleus reticularis thalami neurons determine the characteristic firing patterns of these neurons. We show that subtle modifications in T channel gating that are at the limit of the resolution achieved in classical biophysical studies in heterologous expression systems have profound consequences for synaptically evoked firing dynamics in native neurons. Moreover, we demonstrate that the biophysical properties of the T current in the voltage region corresponding to the foot of the activation and inactivation curves drastically condition physiologically evoked burst firing with a high degree of synaptic input specificity.

The properties of reticular thalamic neuron GABA(A) IPSCs of absence epilepsy rats lead to enhanced network excitability.

Both human investigations and studies in animal models have suggested that abnormalities in GABA(A) receptor function have a potential role in the pathophysiology of absence seizures. Recently we showed that, prior to seizure onset, GABA(A) IPSCs in thalamic reticular (NRT) neurons of genetic absence epilepsy rats from Strasbourg (GAERS) had a 25% larger amplitude, a 40% faster decay and a 45% smaller paired-pulse depression than those of nonepileptic control (NEC) rats. By means of a novel mathematical description, the properties of both GAERS and NEC GABAergic synapses can be mimicked. These model synapses were then used in an NRT network model in order to investigate their potential impact on the neuronal firing patterns. Compared to NEC, GAERS NRT neurons show an overall increase in excitability and a higher frequency and regularity of firing in response to periodic input signals. Moreover, in response to randomly distributed stimuli, the GAERS but not the NEC model produces resonance between 7 and 9 Hz, the frequency range of spike-wave discharges in GAERS. The implications of these results for the epileptogenesis of absence seizures are discussed.

Modulation of neuronal T-type calcium channels.

As T-type calcium channels open near resting membrane potential and markedly influence neuronal excitability their activity needs to be tightly regulated. Few neuronal T-current regulations have been described so far, but interestingly some of them involve unusual mechanisms like G protein-independent but receptor-coupled modulation, while the use of recombinant channels has established both a direct action of Gbetagamma subunits, anandamide, arachidonic acid and a phosphorylation process by CaMKII. Nearly all reported types of modulation involve Cav3.2 channels while no regulation of Cav3.1 has been reported, a difference that may originate from diversities in the intracellular loop connecting the II and III domains of the two isotypes. The search for T-current regulators requires taking into account their peculiar activation properties, since a close link may exist between the channel conformation and its modulation. Indeed, in thalamocortical neurons a phosphorylation-mediated regulation of the amplitude of the T-current has been shown to be highly dependent upon the state of the channel and only to become apparent when the channels are in the voltage range close to neuronal resting membrane potential.

Cortical-area specific block of genetically determined absence seizures by ethosuximide.

Absence epilepsy is characterised by a paroxysmal loss of consciousness, of abrupt onset and termination, and is associated with a bilateral synchronous spike and wave discharge (SWD) on the electroencephalogram. Absence seizures involve an interplay between thalamic and cortical structures, although most research has so far focussed on sensory thalamic nuclei and the reticular thalamic nucleus (RTN). Thus, microinfusion of ethosuximide (ETX), a first choice anti-absence drug, into either the ventrobasal thalamus or RTN of the genetic absence epilepsy rat from Strasbourg (GAERS), a validated rat model of absence epilepsy, does not produce immediate cessation of seizure activity, as is seen following systemic administration. As recent evidence indicates a seizure initiation site within the peri-oral region of the primary somatosensory cortex (S1po), we have now applied ETX into S1po as well as the somatosensory cortex forelimb region (S1FL) and the motor cortex (M1) of freely moving GAERS. Microinfusion of 10 or 20 nmol/side of ETX into S1po produced an immediate cessation of seizure activity. A less marked response was produced when even a higher dose (200 nmol/side) was infused into S1FL. No reduction of SWD was seen when ETX was infused into M1. Microinfusion of CGP 36742 (5 nmol/side), a GABA(B) antagonist, produced immediate cessation of seizure activity in both S1po and M1 and a delayed effect in S1FL. These data suggest that the ability of ETX to abolish genetically determined absence seizures is cortical-area specific and support the involvement of S1po in the initiation of SWDs.

Somatostatin inhibits GABAergic transmission in the sensory thalamus via presynaptic receptors.

The action of somatostatin on GABA-mediated transmission was investigated in cat and rat thalamocortical neurons of the dorsal lateral geniculate nucleus and ventrobasal thalamus in vitro. In the cat thalamus, somatostatin (10 microM) had no effect on the passive membrane properties of thalamocortical neurons and on the postsynaptic response elicited in these cells by bath or iontophoretic application of (+/-)baclofen (5-10 microM) or GABA, respectively. However, somatostatin (1-10 microM) decreased by a similar amount (45-55%) the amplitude of electrically evoked GABA(A) and GABA(B) inhibitory postsynaptic potentials in 71 and 50% of neurons in the lateral geniculate and ventrobasal nucleus, respectively. In addition, the neuropeptide abolished spontaneous bursts of GABA(A) inhibitory postsynaptic potentials in 85% of kitten lateral geniculate neurons, and decreased (40%) the amplitude of single spontaneous GABA(A) inhibitory postsynaptic potentials in 87% of neurons in the cat lateral geniculate nucleus. Similar results were obtained in the rat thalamus. Somatostatin (10 microM) had no effect on the passive membrane properties of thalamocortical neurons in this species, or on the outward current elicited by puff-application of (+/-)baclofen (5-10 microM). However, in 57 and 22% of neurons in the rat lateral geniculate and ventrobasal nuclei, respectively, somatostatin (1 microM) reduced the frequency, but not the amplitude, of miniature GABA(A) inhibitory postsynaptic currents by 31 and 37%, respectively. In addition, the neuropeptide (1 microM) decreased the amplitude of evoked GABA(A) inhibitory postsynaptic currents in 20 and 55% of rat ventrobasal neurons recorded in normal conditions and during enhanced excitability, respectively: this effect was stronger on bursts of inhibitory postsynaptic currents(100% decrease) than on single inhibitory postsynaptic currents (41% decrease). These results demonstrate that in the sensory thalamus somatostatin inhibits GABA(A)- and GABA(B)-mediated transmission via a presynaptic mechanism, and its action is more prominent on bursts of GABAergic synaptic currents/potentials.

On the putative contribution of GABA(B) receptors to the electrical events occurring during spontaneous spike and wave discharges.

Cortical and thalamic neurones play a major role in the generation/expression of spike and wave discharges (SWDs), the main electroencephalographic (EEG) feature of absence seizures. The detailed mechanisms leading to this paroxysmal EEG activity, however, are still poorly understood. We have now made in vivo intracellular recordings from layer V cortical neurones of the facial motor cortex and from thalamocortical (TC) neurones of the ventroposteromedial and ventroposterolateral nuclei in a well established model of this disease: the Genetic Absence Epilepsy Rats from Strasbourg (GAERS). The main feature of the intracellularly recorded activity of TC neurones during spontaneous SWDs was the presence of rhythmic sequences of synaptic potentials consisting of an EPSP closely followed by 2-6 IPSPs. These rhythmic sequences were superimposed on a small tonic hyperpolarization that lasted for the whole duration of the SWD and was still present at potentials close to -85 mV. The rhythmic IPSPs, on the other hand, had a reversal potential of -68 mV, and always appeared as depolarizing events when recording with KCl-filled electrodes at -55 mV. Low frequency electrical stimulation of the corresponding cortical area evoked in TC neurones a short and a long lasting IPSP, whose waveforms were reminiscent of a GABA(A) and a GABA(B) IPSP, respectively. The main feature of the intracellular activity recorded in cortical neurones during spontaneous SWDs was the presence of rhythmic depolarizations. Their frequency was similar to the one of SWDs in the EEG, and was not affected by DC injection. The amplitude of the rhythmic depolarizations, however, increased following steady hyperpolarization of the neurone by DC injection. An increase in the apparent input resistance of cortical neurones was observed during SWDs compared to the inter-SWDs periods. Low frequency electrical stimulation of the contralateral striatum evoked in cortical neurones a short and a long lasting IPSP, whose waveforms were reminiscent of a GABA(A) and a GABA(B) IPSP, respectively. Our data indicate that there are no rhythmic GABA(B) IPSPs and low threshold Ca2+ potentials in GAERS TC neurones during SWDs, but rhythmic sequences of EPSP/IPSPs superimposed on a tonic hyperpolarization that might represent a long lasting GABA(B) IPSP. Further experiments are required to clarify the nature of the voltage waveform and the increase in input resistance observed in cortical neurones during spontaneous SWDs in GAERS.

On the action of the anti-absence drug ethosuximide in the rat and cat thalamus.

The action of ethosuximide (ETX) on Na+, K+, and Ca2+ currents and on tonic and burst-firing patterns was investigated in rat and cat thalamic neurons in vitro by using patch and sharp microelectrode recordings. In thalamocortical (TC) neurons of the rat dorsal lateral geniculate nucleus (LGN), ETX (0.75-1 mM) decreased the noninactivating Na+ current, INaP, by 60% but had no effect on the transient Na+ current. In TC neurons of the rat and cat LGN, the whole-cell transient outward current was not affected by ETX (up to 1 mM), but the sustained outward current was decreased by 39% at 20 mV in the presence of ETX (0.25-0.5 mM): this reduction was not observed in a low Ca2+ (0.5 mM) and high Mg2+ (8 mM) medium or in the presence of Ni2+ (1 mM) and Cd2+ (100 microM). In addition, ETX (up to 1 mM) had no effect on the low-threshold Ca2+ current, IT, of TC neurons of the rat ventrobasal (VB) thalamus and LGN and in neurons of the rat nucleus reticularis thalami nor on the high-threshold Ca2+ current in TC neurons of the rat LGN. Sharp microelectrode recordings in TC neurons of the rat and cat LGN and VB showed that ETX did not change the resting membrane potential but increased the apparent input resistance at potentials greater than -60 mV, resulting in an increase in tonic firing. In contrast, ETX decreased the number of action potentials in the burst evoked by a low-threshold Ca2+ potential. The frequency of the remaining action potentials in a burst also was decreased, whereas the latency of the first action potential was increased. Similar effects were observed on the burst firing evoked during intrinsic delta oscillations. These results indicate an action of ETX on INaP and on the Ca2+-activated K+ current, which explains the decrease in burst firing and the increase in tonic firing, and, together with the lack of action on low- and high-threshold Ca2+ currents, the results cast doubts on the hypothesis that a reduction of IT in thalamic neurons underlies the therapeutic action of this anti-absence medicine.

Intracellular recordings in thalamic neurones during spontaneous spike and wave discharges in rats with absence epilepsy.

1. In vivo extracellular and intracellular recordings were performed from thalamocortical (TC) neurones in a genetic model of absence epilepsy (genetic absence epilepsy rats from Strasbourg) during spontaneous spike and wave discharges (SWDs). 2. Extracellularly recorded single units (n = 14) fired either a single action potential or a high frequency burst of up to three action potentials, concomitantly with the spike component of the spike-wave complex. 3. Three main events characterized the intracellular activity of twenty-six out of twenty-eight TC neurones during SWDs: a small amplitude tonic hyperpolarization that was present throughout the SWD, rhythmic sequences of EPSP/IPSPs occurring concomitantly with the spike-wave complexes, and a small tonic depolarization at the end of the SWD. The rhythmic IPSPs, but not the tonic hyperpolarization, were mediated by activation of GABAA receptors since they reversed in polarity at -68 mV and appeared as depolarizing events when recording with KCl-filled electrodes. 4. The intracellular activity of the remaining two TC neurones consisted of rhythmic low threshold Ca2+ potentials, with a few EPSP/IPSP sequences present at the start of the SWD. 5. These results obtained in a well-established genetic model of absence epilepsy do not support the hypothesis that the intracellular activity of TC neurones during SWDs involves rhythmic sequences of GABAB IPSPs and low threshold Ca2+ potentials.

GABAA receptor-mediated IPSCs in rat thalamic sensory nuclei: patterns of discharge and tonic modulation by GABAB autoreceptors.

1. The patterns of discharge of spontaneous GABAA-mediated inhibitory postsynaptic currents (sIPSCs), originating from the nucleus reticularis thalami (NRT), and their modulation by GABAB autoreceptors, were studied in rat thalamocortical (TC) neurones using whole-cell voltage-clamp recordings in brain slices. 2. sIPSCs were recorded in all ventro-basal (VB) and dorsal lateral geniculate (LGN) neurones. In VB neurones, in the presence of tetraethylammonium (TEA, 5 mM), these sIPSCs can occur in bursts at frequencies of either 0.1 or 1-2 Hz. In the presence of tetrodotoxin (TTX), these bursting activities are replaced by the continuous discharge of miniature IPSCs (mIPSCs), recorded in the absence of TEA, at a frequency of 4 Hz. The kinetic properties of mIPSCs were similar in VB and LGN TC neurones. 3. In VB TC neurones the GABAB receptor agonist (+/-)-baclofen, at a concentration of 0.05 microM, decreased the mIPSC frequency by 22% without affecting their amplitude distribution. Increasing the (+/-)-baclofen concentration to 1 and 10 microM caused similar reductions (41 and 47%, respectively) in the mIPSCs frequency: these values were significantly different from the one observed with 0.05 microM (+/-)-baclofen. In LGN TC neurones, where mIPSCs originate from both NRT and local interneurone terminals, 1 microM (+/-)-baclofen produced a 66% reduction in the mIPSC frequency. 4. The GABAB receptor antagonist CGP55845A (50 nM) not only blocked the baclofen-mediated decrease in mIPSC frequency, but also produced a 52% increase in the mIPSC frequency compared with control in three out of seven neurones. Application of CGP55845A (50-500 nM) alone produced a 77% increase in the mIPSC frequency in three out of nine VB neurones, and in the LGN, CGP55845A (100 nM) produced a 53% increase in four out of nine neurones. CGP55845A (100 nM) also reversibly increased the amplitude of evoked GABAA IPSCs by 74 and 57% in three out of three VB and three out of five LGN neurones, respectively. 5. Application of GABA (1.5-5 microM) decreased the mIPSC frequency in VB TC neurones by a similar extent (48%) as 1-10 microM (+/-)-baclofen. 6. In the presence of 100 microM Cd2+, (+/-)-baclofen still decreased the mIPSC frequency by about 40%, indicating that the effect of presynaptic GABAB receptor activation on spontaneous GABA release did not occur through a reduction of voltage-dependent Ca2+ currents. 7. Cd2+ (100 microM) decreased the amplitude of both mIPSCs and isoguvacine-induced current by 30 and 19%, respectively, indicating an effect of this divalent cation on postsynaptic GABAA receptors. 8. We conclude that GABAB autoreceptors are present on the GABAergic terminals within the thalamic sensory nuclei and that these receptors can be tonically activated by the ambient GABA.

Modulation by different GABAB receptor types of voltage-activated calcium currents in rat thalamocortical neurones.

1. The effects of the GABAB receptor agonist baclofen on the voltage-dependent Ca2+ currents were studied in rat thalamocortical neurones with the use of whole cell voltage-clamp recordings in brain slices. 2. The contribution of N-, L- and P-types of Ca2+ channels to the total high voltage-activated Ca2+ (HVA Ca2+) current was assessed by the use of omega-conotoxin, nifedipine and omega-agatoxin IVA, respectively. No P-type current could be detected. Thus, the HVA Ca2+ current contained an N- and an L-type current (23 and 15% of the total current, respectively) and a residual current, which will be referred to as the 'R' component. 3. Baclofen (1-50 microM) had no effect on the low voltage-activated (LVA) Ca2+ current (IT). 4. At low concentrations (0.5-10 microM), baclofen decreased the HVA Ca2+ currents by about 10-20% without a marked modification on the kinetics, whereas 50 microM baclofen decreased the HVA Ca2+ currents by about 40% with a pronounced slowing down of the kinetics. 5. The 10-20% decrease of the total HVA Ca2+ currents produced by the low concentrations of baclofen occurred as the result of a 30% block of the 'R' component. The additional decrease observed with the dose of 50 microM was due to a full block of the N-type current. The L-type was unaffected by baclofen. 6. The effect of baclofen on the total HVA Ca2+ current was partially blocked by GABAB receptor antagonists indicating that it occurred through stimulation of GABAB receptors. 7. The effect of baclofen on the N-type current was abolished by CGP 35348 (100 microM) and CGP 55845A (100 nM). The effect on the 'R' component was also antagonized by CGP 55845A (100 nM) although with a lower potency, but was not blocked by CGP 35348 (100 microM). 8. We conclude that the effects of baclofen on the various components of the HVA Ca2+ currents occur through different types of GABAB receptors. One receptor has a high affinity for baclofen (i.e. saturated by concentrations as low as 0.5 microM), is insensitive to CGP 35348, is coupled to the 'R' component and is responsible for a maximum 20% decrease in the total HVA Ca2+ current. The other receptor has a lower affinity for baclofen (i.e. affected by a concentration of 50 microM), is sensitive to CGP 35348, is coupled to the N-type Ca2+ current and is responsible for the additional 20-30% decrease in the HVA Ca2+ current observed with 50 microM baclofen.

Sensory input and burst firing output of rat and cat thalamocortical cells: the role of NMDA and non-NMDA receptors.

1. Intracellular and patch-clamp recordings were obtained from thalamocortical (TC) cells in the rat and cat dorsal lateral geniculate nucleus (dLGN) in vitro to study the role of N-methyl-D-aspartate (NMDA) and non-NMDA receptors in the synaptic potential and burst firing evoked by electrical stimulation of the optic tract. 2. At membrane potentials more positive than -65 mV, the sensory synaptic potential consisted of a fast EPSP that was followed by a smaller, slower component. At membrane potentials more negative than -65 mV, this slower component became more prominent owing to the presence of a low-threshold (LT) Ca2+ potential, which in turn evoked a high-frequency (> 150 Hz) burst of action potentials. The lower, but not the upper limit of the range of membrane potential over which burst firing occurred was dependent on the amplitude of the fast EPSP. 3. The non-NMDA receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 5-10 microM) and 1-(4-amino-phenyl)-4-methyl-7,8-methylene-dioxy-5H-2,3- benzodiazepine (GYKI 52466, 100 microM) greatly depressed the fast EPSP, abolished the burst firing generated by the LT Ca2+ potential, and left a relatively small, slow EPSP, which was sensitive to the NMDA antagonist DL-2-amino-5-phosphonovaleric acid (DL-AP5, 50-100 microM). 4. In the absence of CNQX or GYKI 52466, DL-AP5 depressed the slow but not the fast EPSP. DL-AP5 also increased the latency of the first action potential evoked by the LT Ca2+ potential or even abolished the LT Ca2+ potential and associated burst firing. The latter effect was only present when this type of firing occurred within a small membrane potential range. 5. DL-AP5 had no effect on the properties of the LT Ca2+ current IT, indicating that its effect on the burst firing was not mediated by a direct action on IT. 6. The response of TC cells to high-frequency (100 Hz) stimulation consisted of an initial burst firing response, followed by a sustained depolarization that could reach firing threshold. This sustained depolarization was markedly depressed by DL-AP5 but not by CNQX. 7. These results demonstrate that with low-frequency stimulation of the sensory afferents, the generation of TC cell output in the rat and cat dLGN is mainly controlled by non-NMDA receptors, while the contribution of NMDA receptors is limited to the burst firing generated by the LT Ca2+ potential, and depends on the membrane potential range over which this type of firing occurs.(ABSTRACT TRUNCATED AT 400 WORDS)

Thalamic low threshold calcium current in a genetic model of absence epilepsy.

The low threshold calcium current (IT) in thalamo-cortical neurones contributes to the generation of spike and wave discharges (SWDs) characteristic of generalized, non-convulsive absence epilepsy. The biophysical properties of this current were analysed in dorsal lateral geniculate neurones from the Genetic Absence Epilepsy Rats from Strasbourg (GAERS+). No difference was found in the voltage dependence and kinetics of IT between GAERS+ and rats of the non epileptic control strain (GAERS-). Thus, a dysfunction of IT does not appear to underlie the occurrence of SWDs in absence epilepsy.

Synaptic Currents in Thalamo-cortical Neurons of the Rat Lateral Geniculate Nucleus.

Thalamo-cortical neurons were identified in slices of the rat dorsal lateral geniculate nucleus and whole-cell currents were recorded using the patch-clamp technique. Postsynaptic currents occurring spontaneously, or elicited by extracellular stimulation in the vicinity of the recorded neuron, were analysed. Spontaneous postsynaptic currents were observed in every recorded neuron. At a holding potential of - 60 mV, and with a high internal Cl-, the currents were inward and had amplitudes ranging from < 10 to 425 pA. All the spontaneous currents were blocked by 10 microM bicuculline, indicating that they were due to the activation of postsynaptic gamma-aminobutyric acid (GABAA) receptors. The 10-90% rise time of these spontaneous GABAergic currents was 0.86 +/- 0.19 ms. Their time course of decay could be fitted to an exponential function with one time constant of 18.19 +/- 3.02 ms (mean +/- SD), or two time constants of 4.47 +/- 0.77 and 33.27 +/- 3.74 ms. This activity was frequently organized in bursts. Stimulus-evoked postsynaptic currents were recorded and shown to be due to the activation of glutamatergic receptors. Under similar experimental conditions a bicuculline-sensitive component was also recorded. These stimulus-evoked GABAergic currents had a 10 - 90% rise time of 1.93 +/- 0.54 ms. Their time course of decay could also be fitted to an exponential function with one time constant of 24.42 ms or two time constants of 10.26 +/- 2.46 and 49.30 +/- 10.98 ms. The difference in the time course between spontaneous and evoked GABAergic currents suggests that these responses may arise from synapses having different locations.

Low-frequency oscillatory activities intrinsic to rat and cat thalamocortical cells.

1. Low-frequency membrane potential oscillations recorded intracellularly from thalamocortical (TC) cells of the rat and cat dorsal lateral geniculate nucleus (dLGN) and of the rat ventrobasal nucleus (VB) maintained in vitro were investigated. On the basis of their electrophysiological and pharmacological properties, four types of activity were distinguished and named: the pacemaker oscillations, the spindle-like oscillations, the 'very slow' oscillations and the 'N-methyl-D-aspartate' (NMDA) oscillations. 2. The pacemaker oscillations (95 out of 173 cells) consisted of rhythmic, large-amplitude (10-30 mV) depolarizations which occurred at a frequency of 1.8 +/- 0.3 Hz (range, 0.5-2.9 Hz) and could often give rise to single or a burst of action potentials. Pacemaker oscillations were observed when the membrane potential was moved negative to -55 and positive to -80 mV, but in a given cell the upper and lower limits of this voltage range were separated by only 13.1 +/- 0.5 mV. Above -45 mV tonic firing consisting of single action potentials was seen in the cells showing this or the other types of low-frequency oscillations. 3. The spindle-like oscillations were observed in thirty-nine (out of 173) TC cells and consisted of rhythmic (2.1 +/- 0.3 Hz), large-amplitude depolarizations (and often associated burst firing) similar to the pacemaker oscillations but occurring in discrete periods every 5-25 s and lasting for 1.5-28 s. The spindle-like oscillations were observed when the membrane potential was moved negative to -55 and positive to -80 mV and in two cells they were transformed into continuous pacemaker oscillations by depolarization of the membrane potential to -60 mV. 4. Pacemaker and spindle-like oscillations were unaffected by tetrodotoxin (TTX) or by selective blockade of NMDA, non-NMDA, GABAA, GABAB, nicotinic, muscarinic, alpha- and beta-noradrenergic receptors. 5. The 'very slow' oscillations consisted of a TTX-insensitive, slow hyperpolarization-depolarization sequence (5-15 mV in amplitude) which lasted up to 90 s and was observed in nine dLGN cells and in two VB cells. The pacemaker and the spindle-like oscillations were recorded in one cell each which also showed the 'very slow' oscillations. 6. The 'NMDA' oscillations were observed only in a 'Mg(2+)-free' medium (0 mM-Mg2+, 2-4 mM-Ca2+; 64 out of 72 cells) and consisted of large-amplitude (10-25 mV) depolarizations that did not occur at regular intervals and were intermixed with smaller depolarizations present on the baseline and on the falling phase of the larger ones.(ABSTRACT TRUNCATED AT 400 WORDS)

Two inward currents and the transformation of low-frequency oscillations of rat and cat thalamocortical cells.

1. The contribution of a slow, mixed Na(+)-K+, inward rectifying current (Ih) and the T-type Ca2+ current (IT) (that underlies low-threshold Ca2+ potentials) to the low-frequency oscillations observed in rat and cat thalamocortical (TC) cells in vitro was studied using current clamp and single-electrode voltage clamp recordings. 2. From a holding potential of -50 mV, voltage steps negative to -60 mV showed the presence of a slow, non-inactivating inward current, Ih. This current was unaffected by Ba2+ (1-4 mM), tetrodotoxin (0.5-1 microM) and TEA (20 mM, n = 6), reversibly blocked by Cs+ (1-3 mM), and its reversal potential (-33.0 +/- 1.2 mV) followed changes in the extracellular Na+ and K+, but not Cl-, concentration. 3. Application of Cs+ (1-3 mM) abolished the pacemaker oscillations (n = 9), while in six cells that did not show any oscillatory activity Cs+ first evoked the spindle-like oscillations that, in the continuous presence of these ions, were then transformed into the pacemaker oscillations before all activities were finally blocked: all these effects were accompanied by a hyperpolarization and a progressive decrease and final blockade of Ih. Cs+ had no effect on the 'N-methyl-D-aspartate' (NMDA) oscillations (n = 5) and Ba2+ (2 mM, n = 8) did not block the pacemaker, the spindle-like and the 'NMDA' oscillations. 4. In ten cells that showed the pacemaker oscillations selective activation of beta-adrenoceptors by 10-50 microM-noradrenaline (in the presence of alpha-noradrenergic antagonists) or by 20 microM-isoprenaline first transformed the pacemaker oscillations into the spindle-like oscillations that, in the continuous activation of beta-receptors, were finally abolished: all these effects were accompanied by a depolarization and a progressive increase of Ih. 5. In TC cells that showed the pacemaker oscillations application of 1-octanol (50-100 microM), an antagonist of T-type Ca2+ currents, reversibly blocked this activity but concomitantly decreased (50%) the cell input resistance (n = 5). Application of Ni2+ (0.2-0.5 mM, n = 13), another antagonist of IT reversibly blocked the pacemaker, the spindle-like and the 'NMDA' oscillations. 7. In cells showing the pacemaker oscillations it was found that the current developing from the most hyperpolarized potential of an oscillation cycle was an inward relaxation whose time course differed from that of Ih evoked at the same potential.(ABSTRACT TRUNCATED AT 400 WORDS)

Calcium entry increases the sensitivity of cerebellar Purkinje cells to applied GABA and decreases inhibitory synaptic currents.

The sensitivity to GABA of Purkinje cells in thin cerebellar slices was examined by recording either spontaneous inhibitory synaptic currents or ionic currents elicited by local GABA applications. The effects of Ca2+ entry induced by depolarizing voltage pulses were opposite for the two types of currents. Currents due to exogenous GABA applications were increased by a train of voltage pulses. This potentiation was transient with an average half recovery period of 3.7 min. Spontaneous synaptic currents were reduced by depolarizing voltage pulses, with a half recovery time of about 20 s. The inhibition was largely explained by a decrease of the frequency of synaptic events, suggesting that the primary location of the effect was presynaptic. Thus, a Ca2+ rise increases the sensitivity of Purkinje cells to GABA and induces a retrograde inhibition of presynaptic terminals. The latter effect may be due to a diffusible Ca2(+)-dependent messenger.

A role for GABAB receptors in excitation and inhibition of thalamocortical cells.

Gamma-aminobutyric acid (GABA) in the thalamus has mainly been associated with the inhibitory modulation of the sensory and cortical flow of information via a 'classical', chloride-dependent, GABAA receptor-mediated action. However, the discovery of a late, long-lasting potassium-dependent inhibitory postsynaptic potential (IPSP) mediated by GABAB receptors present on thalamocortical cells, has allowed new insights into our understanding of the physiological role of this neurotransmitter. In particular, work on the dorsal lateral geniculate nucleus indicates that together with a relatively weak inhibition, GABAB receptor-mediated IPSPs 'prepare' thalamocortical cells for burst firing by activating low-threshold calcium potentials. Thus, GABA in the thalamus can no longer be viewed only as a 'classical' inhibitory transmitter but also as a neuromodulator with a 'priming' role for burst firing excitation. This dual role of GABAB receptors in inhibition and excitation of thalamocortical cells might allow different interpretations of earlier findings in animals and humans, both in healthy and pathological conditions. It will also help to identify new functions for postsynaptic GABAB receptors in other parts of the central nervous system.

Pacemaker-like and other types of spontaneous membrane potential oscillations of thalamocortical cells.

During EEG-synchronized sleep, thalamic activity is characterized by rhythmic oscillations that till recently have been suggested to require the contribution of intra- and extra-thalamic inputs. The present experiments show that thalamocortical (TC) cells, mechanically and pharmacologically isolated from their intra-thalamic, cortical and brainstem inputs, are capable of different types of spontaneous membrane potential oscillations some of which resemble those observed in TC cells of the living animal during EEG-synchronization.

Optic tract stimulation evokes GABAA but not GABAB IPSPs in the rat ventral lateral geniculate nucleus.

The inhibitory postsynaptic potentials (IPSPs) evoked in neurons of the rat ventral geniculate nucleus (vLGN) by electrical stimulation of the optic tract and the action of GABA and baclofen on the same cells were studied using intracellular recording technique in an in vitro slice preparation. A short latency short duration IPSP always followed the monosynaptic excitatory postsynaptic potential (EPSP). This IPSP reversed in polarity at about -65 mV and was reversibly blocked by bicuculline (50 microM) thus indicating that it represents a GABAA receptor-mediated IPSP. No long-lasting IPSP was evoked in vLGN cells by stimulation of the optic tract, while in the same slice, long-lasting GABAB IPSPs were routinely recorded in the dorsal lateral geniculate nucleus. GABA applied by ionophoresis evoked a hyperpolarization that had a reversal potential close to -70 mV and was antagonized by bicuculline. Baclofen hyperpolarized vLGN neurons and its action was reversibly blocked by the selective GABAB antagonist phaclofen (1 mM). In the presence of bicuculline GABA also produced a hyperpolarization that had properties similar to that evoked by baclofen. These results indicate that, although functional GABAA and GABAB receptors are present on vLGN neurons, stimulation of the optic tract evokes only GABAA but not GABAB mediated IPSPs. The lack of long-lasting GABAB IPSPs could explain the absence of long-lasting inhibition observed in vLGN neurons in vivo following stimulation of the optic tract.

On the properties and origin of the GABAB inhibitory postsynaptic potential recorded in morphologically identified projection cells of the cat dorsal lateral geniculate nucleus.

Intracellular recordings were performed from projection cells of the cat dorsal lateral geniculate nucleus in vitro to investigate the properties and origin of optic tract evoked inhibitory postsynaptic potentials mediated by GABAB receptors and their relationship to the physiologically different cell classes present in this nucleus. In all three main laminae of the dorsal lateral geniculate nucleus, stimulation of the optic tract evoked an excitatory postsynaptic potential followed by two inhibitory postsynaptic potentials. The first is a GABAA receptor mediated inhibitory postsynaptic potential since it was blocked by bicuculline, reversed in polarity following intracellular Cl- injection and had a reversal potential similar to the bicuculline sensitive hyperpolarizing effect of GABA. The second is a GABAB receptor mediated inhibitory postsynaptic potential. Its amplitude was not linearly related to membrane potential (maximal amplitude at -60 mV), it decreased when using frequencies of stimulation higher than 0.05 Hz and it was reversibly increased by addition of bicuculline to the perfusion medium. The reversal potential of GABAB inhibitory postsynaptic potentials was dependent on the extracellular K+ concentration but did not change in the presence of bicuculline or when recording with Cl- filled microelectrodes. While GABAA inhibitory postsynaptic potentials always abolished repetitive firing of projection cells, GABAB inhibitory postsynaptic potentials were able to block weak firing but unable to decrease strong activation of projection cells evoked by direct current injection. Optic tract evoked GABAB (as well as GABAA) inhibitory postsynaptic potentials could be recorded in slices which did not include the perigeniculate nucleus, thus indicating that they are generated by the interneurons of the dorsal lateral geniculate nucleus. Using intracellular injection of horseradish peroxidase, we have found that the GABAB inhibitory postsynaptic potentials are present in projection cells showing many different types of neuronal morphologies. In conclusion, GABA released from interneurons in the dorsal lateral geniculate nucleus is capable of evoking an early, short-lasting GABAA and a late, long-lasting GABAB inhibitory postsynaptic potential in projection cells with diverse morphology, indicating that the late inhibition in the dorsal lateral geniculate nucleus can no longer be associated exclusively with the recurrent inhibitory pathway through the perigeniculate nucleus.