6-cyano-7-nitroquinoxaline-2-3-dione and 2-hydroxysaclofen

GABA(B) receptors are believed to play a role in rhythmic activity in the mammalian brain. The aim of our study was to examine the presynaptic and postsynaptic locations of these receptors in the medial septal diagonal band area (MS/DB), an area known to pace the hippocampus theta rhythm. Whole-cell patch recordings were made from parasagittal MS/DB slices obtained from the 16-25 day rat. Neurons were classified into GABAergic and cholinergic subtypes according to previous electrophysiological criteria. Bath application of the GABA(B) receptor agonist baclofen in the presence of tetrodotoxin, and brief tetanic fiber stimulation in the presence of ionotropic receptor antagonists, provided evidence for the presence of postsynaptic GABA(B) receptor transmission to GABAergic but not cholinergic neurons. Bath application of baclofen, at concentrations too low to elicit postsynaptic activity in MS/DB neurons, significantly reduced the amplitudes of stimulus-evoked ionotropic receptor inhibitory postsynaptic potentials (IPSPs) and excitatory postsynaptic potentials (EPSPs) and the paired pulse depression of these evoked potentials. Baclofen also significantly reduced the frequencies but not the amplitudes of miniature inhibitory postsynaptic currents (IPSCs) and excitatory postsynaptic currents (EPSCs), indicating the presence of presynaptic GABA(B) receptors on GABAergic and glutamatergic terminals in the MS/DB. Baclofen, also at a concentration too low to elicit postsynaptic activity, reduced the frequencies and amplitudes of spontaneous IPSCs and EPSCs recorded in the presence of 200-400 nM kainate. Rhythmic compound IPSCs at theta frequencies were recorded under these conditions in some neurons, and these rhythmic compound IPSCs were disrupted by the activation but not by the inhibition of GABA(B) receptors. These results suggest that GABA(B) receptors modulate rather than generate rhythmic activity in the MS/DB, and that this modulatory effect occurs via receptors located on presynaptic terminals.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Animals, Newborn; Baclofen; Bicuculline; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Drug Interactions; Electric Stimulation; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Excitatory Postsynaptic Potentials; GABA Agonists; GABA Antagonists; In Vitro Techniques; Kainic Acid; Membrane Potentials; Neural Inhibition; Neurons; Patch-Clamp Techniques; Phosphinic Acids; Propanolamines; Rats; Rats, Wistar; Receptors, GABA-A; Septum Pellucidum; Sodium Channel Blockers; Tetrodotoxin; Time Factors; Valine

In the mature nervous system excitatory neurotransmission mediated by glutamate is balanced by the inhibitory actions of GABA. However, during early development, GABA acting at the ligand-gated GABAA Cl- channel also exerts excitatory actions. This raises a question as to whether GABA can exert inhibitory activity during early development, possibly by a mechanism that involves activation of the G protein-coupled GABAB receptor. To address this question we used Ca2+ digital imaging to assess the modulatory role of GABAB receptor signaling in relation to the excitatory effects of glutamate during hypothalamic and cortical neuron development. Ca2+ transients mediated by synaptic glutamate release in neurons cultured from embryonic rat were dramatically depressed by the administration of the GABAB receptor agonist baclofen in a dose-dependent manner. The inhibitory effects of GABAB receptor activation persisted for the duration of baclofen administration (>10 min). Preincubation with the Gi protein inhibitor pertussis toxin resulted in a substantial decrease in the inhibitory actions of baclofen, confirming that a Gi-dependent mechanism mediated the effects of the GABAB receptor. Co-administration of the GABAB receptor antagonist 2-hydroxy-saclofen eliminated the inhibitory action of baclofen. Alone, GABAB antagonist application elicited a marked potentiation of Ca2+ transients mediated by glutamatergic neurotransmission, suggesting that tonic synaptic GABA release exerts an inhibitory tone on glutamate receptor-mediated Ca2+ transients via GABAB receptor activation. In the presence of TTX to block action potential-mediated neurotransmitter release, stimulation with exogenously applied glutamate triggered a robust postsynaptic Ca2+ rise that was dramatically depressed (>70% in cortical neurons, >40% in hypothalamic neurons) by baclofen. Together these data suggest both a pre- and postsynaptic component for the modulatory actions of the GABAB receptor. These results indicate a potentially important role for the GABAB receptor as a modulator of the excitatory actions of glutamate in developing neurons.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Baclofen; Bicuculline; Calcium; Cells, Cultured; Cerebral Cortex; Cytarabine; Drug Synergism; Embryo, Mammalian; Excitatory Amino Acid Antagonists; GABA Antagonists; Glutamic Acid; Hypothalamus, Middle; Membrane Potentials; Neurons; Pertussis Toxin; Rats; Rats, Sprague-Dawley; Receptors, GABA-B; Tetrodotoxin; Virulence Factors, Bordetella

We studied the regulation of spontaneous activity in the embryonic (day 10-11) chick spinal cord. After bath application of either an excitatory amino acid (AP-5 or CNQX) and a nicotinic cholinergic (DHbetaE or mecamylamine) antagonist, or glycine and GABA receptor (bicuculline, 2-hydroxysaclofen, and strychnine) antagonists, spontaneous activity was blocked for a period (30-90 min) but then reappeared in the presence of the drugs. The efficacy of the antagonists was assessed by their continued ability to block spinal reflex pathways during the reappearance of spontaneous activity. Spontaneous activity ceased over the 4-5 hour monitoring period when both sets of antagonists were applied together. After application of glycine and GABA receptor antagonists, the frequency of occurrence of spontaneous episodes slowed and became highly variable. By contrast, during glutamatergic and nicotinic cholinergic blockade, the frequency of occurrence of spontaneous episodes initially slowed and then recovered to stabilize near the predrug level of activity. Whole-cell recordings made from ventral spinal neurons revealed that this recovery was accompanied by an increase in the amplitude of spontaneously occurring synaptic events. We also measured changes in the apparent equilibrium potential of the rhythmic, synaptic drive of ventral spinal neurons using voltage or discontinuous current clamp. After excitatory blockade, the apparent equilibrium potential of the rhythmic synaptic drive shifted approximately 10 mV more negative to approximately -30 mV. In the presence of bicuculline, the apparent equilibrium potential of the synaptic drive shifted toward the glutamate equilibrium potential. Considered with other evidence, these findings suggest that spontaneous rhythmic output is a general property of developing spinal networks, and that GABA and glycinergic networks alter their function to compensate for the blockade of excitatory transmission.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; Animals; Baclofen; Bicuculline; Chick Embryo; Cholinergic Antagonists; Dihydro-beta-Erythroidine; Evoked Potentials; Excitatory Amino Acid Antagonists; GABA Antagonists; GABA-A Receptor Antagonists; Glycine; Glycine Agents; Mecamylamine; Motor Neurons; Neuronal Plasticity; Neurotransmitter Agents; Nicotinic Antagonists; Periodicity; Spinal Cord; Strychnine

The concept of dormant interneurons is proving to be hard to define precisely. We argue here that the term is best used as an operational description of interneurons which are not lost from the epileptic brain, but which fail to perform adequately. We present evidence for the existence of functionally dormant interneurons in the tetanus toxin model of chronic epilepsy, and we explore the roles of a partial dormancy (and also of charge-screening) in the acute low magnesium model of epilepsy.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Baclofen; Bicuculline; Computer Simulation; Epilepsy; GABA Antagonists; Hippocampus; In Vitro Techniques; Interneurons; Male; Models, Neurological; Neurons; Pyramidal Cells; Rats; Tetanus Toxin

There is great interest in understanding the mechanisms of expression underlying long-term potentiation (LTP). They are agreed to involve an increase in synaptic efficacy, which is described by three multiplicative parameters: p, the probability of neurotransmitter release; n, the number of active release sites; and q, the postsynaptic unit response to transmitter release. We report three new lines of evidence suggesting that increases in p contribute to LTP expression. (i) When the contributions to LTP by p, n, and q are maximized, and p alone is decreased, another high-frequency stimulation elicits additional LTP. The additional potentiation is only associated with decreases in paired-pulse facilitation (PPF) suggesting an increase in p. (ii) There is an inverse relationship between baseline p [corrected] and the magnitude of LTP elicited, consistent with p [corrected] having more or less room to increase when p is smaller or greater. (iii) It has been shown that there is an inverse relationship between the magnitude of LTP induced and the associated changes in PPF. Now I find that decreasing p before inducing LTP moves the set-point for measuring those changes in PPF from before to after p is decreased, which would only occur if p contributes to LTP. Three lines of evidence, then, suggest that increases in p contribute to LTP expression, which is consistent with a presynaptic contribution to LTP. These experiments do not address potential postsynaptic contributions.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Baclofen; Cadmium Chloride; Calcium Channel Blockers; Evoked Potentials; Hippocampus; In Vitro Techniques; Long-Term Potentiation; Neurotransmitter Agents; omega-Conotoxin GVIA; Peptides; Probability; Rats; Rats, Sprague-Dawley; Receptors, AMPA; Receptors, GABA-B; Receptors, N-Methyl-D-Aspartate; Synapses; Synaptic Transmission; Time Factors

Serotonergic projections are widespread in the developing neocortex, but their functions are obscure. The effects of 5-HT3 receptor agonists on cortical circuit response properties were studied in slices of ferret primary visual cortex using high-speed optical imaging of voltage-sensitive dye signals and whole-cell patch-clamp recording. Activation of the 5-HT3 receptor decreased the amplitude and lateral extent of excitation throughout postnatal development. This effect peaks after eye opening, which indicates a function for serotonergic modulation of circuit responses during the period of refinement of cortical connections. Whole-cell patch-clamp recordings from single neurons revealed that synaptic responses evoked by white matter stimulation were reduced by 5-HT3 receptor agonists, whereas the frequency of spontaneous GABAergic synaptic currents was enhanced dramatically. This indicates that the modulation of spontaneous synaptic activity by fast-acting serotonin receptors is reflected in an inhibition of the circuit response, in line with the notion of background synaptic activity altering the spatiotemporal integration properties of cortical cells by changing their membrane potential and their electrotonic structure. These mechanisms may regulate the response properties of intrinsic circuits in both the adult and developing neocortex.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Animals; Baclofen; Ferrets; gamma-Aminobutyric Acid; Indoles; Ligands; N-Methylaspartate; Neurons; Patch-Clamp Techniques; Pyridinium Compounds; Quinoxalines; Receptor, Serotonin, 5-HT1B; Receptors, GABA-A; Receptors, GABA-B; Receptors, Serotonin; Receptors, Serotonin, 5-HT3; Serotonin; Serotonin Antagonists; Serotonin Receptor Agonists; Synapses; Synaptic Transmission; Tetrodotoxin; Time Factors; Tropisetron; Virulence Factors, Bordetella; Visual Cortex

alpha-Decarboxylation of excitatory amino acids can produce derivatives with depressant actions on the central nervous system. Examples are aspartate-B-alanine and glutamate-GABA. Quisqualate derivatives by alpha-decarboxylation, Quisqualamine (QUAM) and Homoquisqualamine (HOMOQUAM) (with an extra methylene group in the molecular chain), were studied in isolated spinal cord in vitro preparation. Dose-dependent depolarizations and inhibition of spontaneous ventral root potentials (sVRP) were induced by QUAM and HOMOQUAM in unblocked, Mg2+ free, hemisected cord. Ventral root evoked potentials by electrical stimulation of dorsal root (2ms, 30V, pulse/30 sec) (DR-VRP) remained unchanged. In Tetrodotoxin (TTX) medium, HOMOQUAM showed a twofold increment of relative potency respect to QUAM depolarizations. Actions evoked by QUAM and HOMOQUAM were not affected by the addition of N-Methyl-D-Aspartate (NMDA) receptor blockers Mg2+ and DL-AP5; non-NMDA antagonist CNQX and GABA B receptor antagonist 2-hydroxysaclofen. In presence of GABA A receptor blockers Bicuculline MeCl or Picrotoxin, the actions evoked by QUAM and HOMOQUAM were blocked. The results obtained show that GABA A receptor seemed to mediate QUAM and HOMOQUAM activity in spinal cord in vitro preparation. The addition of a methylene group in the molecular chain increased the potency twice although the kinetic of the drug did not appeared to have changed. The development of new compounds with depressant activity in the central nervous system may be useful in assessing the physiological and therapeutic significance of central GABA receptors, especially if the blockade of spontaneous activity is not followed by an alteration of the neuronal integration and synaptic transmission reflected in the DR-VRP.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Animals; Baclofen; Bicuculline; Decarboxylation; Evoked Potentials; GABA Agonists; Magnesium; Muscimol; Picrotoxin; Quisqualic Acid; Rats; Rats, Wistar; Receptors, GABA; Receptors, N-Methyl-D-Aspartate; Spinal Cord; Tetrodotoxin

1. Tight seal, whole-cell recordings from auditory cortex in vivo and in vitro were obtained to investigate modification of N-methyl-D-aspartate (NMDA) receptor-mediated synaptic activity by paired-pulse afferent stimulation. 2. In recordings from urethane-anaesthetized rats (at 37 degrees C), or from cortical slices maintained in vitro (32 degrees C), afferent stimulation elicited a monosynaptic early EPSP and polysynaptic early and late IPSPs. In addition, a late EPSP could be elicited when the stimulus was preceded by an identical priming stimulus (interval approximately 200 ms). The late EPSP was attenuated by the NMDA receptor antagonist DL-2-amino-5-phosphonovalerate (APV, 50 microM). 3. Bath application of the gamma-aminobutyric acid-B (GABAB) receptor antagonist 3-amino-2-(4-chlorophenyl)-2-hydroxy-propylsulphonic acid (2-OH-saclofen; 50 microM) attenuated the late IPSP and clearly revealed a late EPSP. However, 2-OH-saclofen had lesser effects on the second late EPSP elicited during paired-pulse stimulation. Membrane depolarization in 2-OH-saclofen increased the magnitude of the early IPSP, which suppressed the late EPSP once again. Since pharmacological blockade of EPSPs revealed paired-pulse depression of monosynaptically elicited early and late IPSPs, these data indicate that (1) both early and late IPSPs were capable of suppressing the late EPSP, and (2) these effects were reduced during paired-pulse stimulation. 4. Pharmacological isolation of the late EPSP allowed testing of the direct effect of paired-pulse stimulation. Application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 20 microM), picrotoxin (10 microM) and 2-OH-saclofen (50 microM) isolated the late EPSP (onset, 3 ms; peak latency, 28 ms; peak amplitude, 7 mV; duration, 240 ms), which grew in magnitude with membrane depolarization and was largely (> 90%) blocked by APV. Paired-pulse stimulation depressed the isolated late EPSP by 30%. 5. Thus, apparent paired-pulse facilitation of the late EPSP is attributable to release from GABAergic inhibition, and not to direct facilitation. Facilitation of the late EPSP is a functional consequence of IPSP depression. The results indicate the importance of inhibition in regulating synaptic activity mediated by NMDA receptors.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Auditory Cortex; Baclofen; Cerebral Cortex; Cesium; Electric Stimulation; Electrophysiology; Evoked Potentials; GABA Antagonists; gamma-Aminobutyric Acid; In Vitro Techniques; Male; Membrane Potentials; Patch-Clamp Techniques; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Synapses; Thalamus

Anatomic and physiologic studies in the rat have shown projections from the hippocampal formation (HF) and mediodorsal (MD) thalamic nucleus to the medial prefrontal cortex (mPFC). The authors used multi-barrel iontophoresis to: confirm the neurotransmitter used in the projection from HF to mPFC; investigate the role of GABAergic inhibition in the regulation of this projection; and examine the functional convergence of projections from HF and MD onto single mPFC neurons. During HF stimulation, nine cells (6%) showed excitation followed by prolonged inhibition, 39 cells (26%) showed prolonged inhibition alone and 100 cells (68%) showed no clear response. In a further 12 cells that showed no predrug excitation to HF stimulation (representing 16% of the cells in this category), iontophoresis of the GABAA antagonist bicuculline methiodide (BMI) revealed excitatory responses. A total of six mPFC cells (38% of the cells showing excitatory responses to HF stimulation) showed convergent excitation to HF and MD thalamic (or adjacent paratenial nucleus) stimulation. Five out of eight (63%) of the predrug or BMI-revealed excitatory responses of mPFC neurons to HF stimulation were selectively decreased after AMPA antagonist iontophoresis (either CNQX or DNQX). These data confirm that the HF projection to prefrontal cortex is, at least in part, glutamatergic; suggest that the responses of mPFC neurons to activity in this HF pathway are regulated by GABAergic inhibition; and indicate that projections from HF and MD converge onto single mPFC neurons.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; Animals; Baclofen; Bicuculline; Brain Mapping; Electric Stimulation; Electrophysiology; GABA-A Receptor Antagonists; GABA-B Receptor Antagonists; gamma-Aminobutyric Acid; Glutamic Acid; Limbic System; Male; Neurons; Organophosphorus Compounds; Piperazines; Prefrontal Cortex; Quinoxalines; Rats; Rats, Sprague-Dawley; Receptors, AMPA; Receptors, N-Methyl-D-Aspartate; Thalamus

Many studies suggest that the dentate gyrus (DG) is a control point for hippocampal epileptogenesis. However, the importance of GABAergic inhibition in the DG is not quite clear. Intracellular recordings were obtained from granule cells (GC) of the rat DG. In addition to GABAA-mediated spontaneous postsynaptic potentials (PSPs), some GC exhibited spontaneous slow hyperpolarizations (SH). The SH were more commonly observed in a high concentration of external potassium. 2-Hydroxysaclofen, a GABAB antagonist, reduced the SH. Focal stimulation of the perforant path (PP) in the subiculum with a single pulse evoked a depolarization followed by a SH, which were both abolished by the excitatory amino acid (EAA) blockers, 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX) and 2-amino-5-phosphonovaleric acid (APV). When evoked with a train of pulses, the SH was unaffected by the EAA blockers in 40% of the cells, suggesting either the existence of a GABAergic PP, or an unidentified polysynaptic mechanism. In control, the synaptic response to PP stimulation was superficially similar whether the stimulus was applied in the subiculum or stratum moleculare. However, in presence of bicuculline, the subicular PSP was followed by a train of PSPs occurring at a constant frequency of 25 Hz. This 'reverberating' effect of bicuculline was decreased in presence of APV and was abolished in slices in which the excitatory transmission had been interrupted downstream from CA3 neurons, suggesting that reverberation required the integrity of the hippocampo-entorhinal loop. By contrast, bicuculline decreased the amplitude of the stratum moleculare PSP. It is concluded that GC receive tonic inhibition from GABA acting at GABAA and GABAB receptors. The role of GABAB receptors is unclear; by contrast, GABAA-mediated inhibition prevents GC from reverberated excitation. The probability of occurrence of reverberation is higher during activation of the whole temporo-ammonic pathway and is partly dependent on the activation of N-methyl-D-aspartate (NMDA) receptors. Thus, the in vitro brain slice can be used as a model to study reverberation which has been recently demonstrated to underlie epileptiform discharges in the whole brain preparation.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Baclofen; Bicuculline; Electric Stimulation; Epilepsy; Evoked Potentials; gamma-Aminobutyric Acid; Hippocampus; In Vitro Techniques; Male; Membrane Potentials; Pyramidal Tracts; Quinoxalines; Rats; Rats, Sprague-Dawley; Receptors, GABA-A; Receptors, GABA-B; Synapses

In humans temporal lobe epilepsy (TLE) is characterized by recurrent seizures, neuronal hyperexcitability, and selective loss of certain neuronal populations in the hippocampus. Animal models of the condition indicate that a diminution of inhibition mediated by gamma-aminobutyric acid (GABA) accounts for the altered function, and it has been hypothesized that the diminution arises because GABAergic basket interneurons are "dormant" as a result of their being disconnected from excitatory inputs. In hippocampal slices, inhibitory postsynaptic potentials (IPSPs) were elicited in CA1 pyramidal cells by activation of basket cells; responses from an animal model of TLE were compared to those from control tissue. IPSPs evoked indirectly by activation of terminals that then excited basket cells were reduced in the epileptic tissue, whereas IPSPs evoked by direct activation of basket cells, when excitatory neurotransmission was blocked, were not different from controls. These results provide support for the "dormant basket cell" hypothesis and have implications for the pathophysiology and treatment of human TLE.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; Animals; Baclofen; Electric Stimulation; Epilepsy, Temporal Lobe; Evoked Potentials; Hippocampus; In Vitro Techniques; Interneurons; Male; Membrane Potentials; Picrotoxin; Pyramidal Tracts; Quinoxalines; Rats; Rats, Sprague-Dawley; Receptors, GABA-A; Receptors, N-Methyl-D-Aspartate; Status Epilepticus

To examine the mechanisms underlying chronic epileptiform activity, field potentials were first recorded to identify hyperexcitable hippocampal slices from kainic acid-treated rats. Intracellular recordings were then obtained from CA1 pyramidal cells in the hyperexcitable areas. Twenty-two of the 47 cells responded to electrical stimulation of the stratum radiatum with a burst of two or more action potentials and reduced early inhibitory postsynaptic potentials, and were considered hyperexcitable. The remaining 25 cells were not hyperexcitable, displaying a single action potential and biphasic inhibitory postsynaptic potentials after stimulation, like control cells (n = 20). A long duration, voltage-sensitive component was associated with subthreshold excitatory postsynaptic potentials in the majority of hyperexcitable (12/15) and non-hyperexcitable (3/5) cells examined from kainic acid-treated animals, but not from cells (1/10) of control animals. Stimulation of stratum radiatum during pharmacological blockade of ionotropic excitatory amino acid synaptic transmission elicited biphasic monosynaptic inhibitory postsynaptic potentials in all hyperexcitable (n = 9) and non-hyperexcitable (n = 9) cells tested from kainate-treated animals, as well as in control cells (n = 8). The mean amplitude, latency to peak, equilibrium potential, and conductance changes of early and late monosynaptic inhibitory postsynaptic potentials were not different between cells of kainic acid-treated and control animals. In seven hyperexcitable cells tested, the early component of monosynaptic inhibitory postsynaptic potentials was significantly reduced by the GABAA receptor antagonist bicuculline (100-200 microM). The late component was significantly decreased by the GABAB receptor antagonist 2-hydroxysaclofen (1-2 mM; n = 3). Comparable effects were observed on early and late monosynaptic inhibitory postsynaptic potentials in non-hyperexcitable cells (n = 4) from kainic acid-treated animals and control cells (n = 5). These results suggest that GABAergic synapses on hyperexcitable hippocampal pyramidal cells of kainate-treated rats are intact and functional. Therefore, epileptiform activity in the kainate-lesioned hippocampus may not arise from a disconnection of GABAergic synapses made by inhibitory interneurons on pyramidal cells. The hyperexcitability may be due to underactivation of inhibitory interneurons and/or reorganization of excitatory inputs to pyramidal cells since, in

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Baclofen; Evoked Potentials; gamma-Aminobutyric Acid; Hippocampus; In Vitro Techniques; Kainic Acid; Neurons; Pyramidal Tracts; Quinoxalines; Rats; Rats, Sprague-Dawley; Receptors, GABA-A; Synapses

Injecting 0.5-1.0 microgram of cholera toxin into rat hippocampus induces a chronic epileptic focus which generates interictal discharges and brief epileptic seizures intermittently over the following seven to 10 days. Here we examined the electrophysiological properties of hippocampal slices prepared from these rats three to four days after injection, at the height of the epileptic syndrome. These slices generated epileptic discharges in response to electrical stimulation of afferent pathways. In many cases epileptic discharges occurred spontaneously in the CA3 subregion; these usually lasted < 200 ms, but they could last < 0.6 s. Intracellular recordings from pyramidal layer cells revealed depolarization shifts synchronous with the epileptic field potentials. These depolarization shifts had slow onsets compared with those induced by blocking inhibition with bicuculline (depolarizations started a mean of 57 ms before, and reached 5.2 mV by, the onset of the cholera toxin epileptic field potential, compared with 12 ms and 3.6 mV respectively for 70 microM bicuculline methiodide). Extracellular unit recordings showed that the slow predepolarization seen in the cholera toxin focus was associated with an acceleration of the firing of other pyramidal layer neurons. The epileptic activity in this model cannot be attributed to the loss of synaptic inhibition, because inhibitory postsynaptic potentials could be evoked when the synchronous bursts were blocked by increasing [Ca2+]o from 2 to 8 mM. Observations of monosynaptic inhibitory postsynaptic currents isolated by application of 20 microM 6-cyano-7-nitroquinoxaline-2,3-dione, 50 microM DL-2-amino-5-phosphonovaleric acid and 100-200 microM 3-amino-2-(4-chlorophenyl)-2-hydroxy-propylsulphonic acid showed a small effect of the toxin only on the time course of the inhibitory postsynaptic current. On the other hand, there were significant changes in the intrinsic properties of individual neurons. The membrane potentials of cells in the cholera toxin focus did not differ from those in slices from rats injected with vehicle solution, but their input resistances were significantly increased. Unlike the other cellular changes in this model, the increase in input resistance was not seen in slices exposed acutely to 1 micrograms/ml cholera toxin for 30 min, suggesting there may be morphological changes in the chronic focus. Action potential accommodation and the slow afterhyperpolarization were depressed in both acute

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Action Potentials; Animals; Baclofen; Bicuculline; Cholera Toxin; Chronic Disease; Epilepsy; Hippocampus; Injections; Male; Neurons; Quinoxalines; Rats; Rats, Wistar; Receptors, GABA-A; Synaptic Transmission

1. Intracellular recording techniques were used to characterize synaptic inhibitory postsynaptic potentials (IPSPs) recorded from neurons of the basolateral nucleus of the amygdala (BLA). Bipolar electrodes positioned in the stria terminalis (ST) or lateral amygdala (LA) were used to evoke synaptic responses at a frequency of 0.25 Hz. 2. Two synaptic waveforms having IPSP components could be evoked by electrical stimulation of either pathway: a biphasic, excitatory postsynaptic potential (EPSP), fast-IPSP (f-IPSP) waveform, and a multiphasic, EPSP, f-IPSP, and subsequent slow-IPSP (s-IPSP) waveform. Expression of either waveform was dependent on the site of stimulation. ST stimulation evoked a similar number of biphasic (45%) and multiphasic (50%) synaptic responses. In contrast, stimulation of the LA pathway evoked mainly (80%) multiphasic synaptic responses. 3. Both the f- and s-IPSP elicited by ST stimulation could be reduced in amplitude in the presence of the glutamatergic, N-methyl-D-aspartate (NMDA) antagonist, (DL)-2-amino-5-phosphonovaleric acid (APV, 50 microM), and were abolished by the glutamatergic, non-NMDA antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM). In contrast, a CNQX-resistant f-IPSP was evoked with LA stimulation and abolished by subsequent addition of bicuculline methiodide (BMI), a gamma-aminobutyric acid (GABAA) receptor antagonist, suggesting direct inhibition of BLA neurons by GABAergic LA interneurons. The sensitivity of the s-IPSPs and the f-IPSPs to glutamatergic antagonists suggests the presence of feed-forward inhibition onto BLA neurons. 4. The f-IPSP possessed characteristics of potentials mediated by GABAA receptors linked to Cl- channels, namely, a reversal potential of -70 mV, a decrease in membrane resistance (13.5 M omega) recorded at -60 mV, a block by BMI, and potentiation by sodium pentobarbital (NaPB). 5. The s-IPSP was associated with a resistance decrease of 4.5 M omega, a reversal potential of -95 mV, and was reversibly depressed (approximately 66%) by 2-hydroxy-saclofen (100 microM), suggesting activation of GABAB receptors. 6. The large resistance change associated with the f-IPSP, its temporal overlap with evoked EPSPs, and the development of both spontaneous and evoked burst firing in the presence of BMI suggests that the f-IPSP determines the primary state of excitability in BLA neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Amygdala; Animals; Baclofen; Bicuculline; Evoked Potentials; gamma-Aminobutyric Acid; In Vitro Techniques; Membrane Potentials; Neural Inhibition; Pentobarbital; Potassium; Quinoxalines; Rats; Rats, Inbred Strains; Receptors, GABA-A; Receptors, N-Methyl-D-Aspartate; Synapses; Synaptic Transmission

Extracellular recordings of field potentials in area CA1 of the rat hippocampal slice have been used to investigate paired-pulse facilitation. Field potentials were evoked by maximal stimulation of the Schaffer collateral/commissural fibres. The height of the population spike (PS) in stratum pyramidale (str. pyr.) and the area under the field excitatory postsynaptic potential (EPSP) following the PS in the stratum radiatum (str. rad.) were quantified. These values were used to describe the time course of paired-pulse facilitation. Facilitation of the PS was maximal 50 ms after the conditioning pulse and was present over a period of about 500 ms. However, facilitation of the late area (LA) of the field EPSP was maximal afer 125 ms and had an overall duration of 1-2 s. The N-methyl-D-aspartate (NMDA) receptor antagonist, 2-amino-5-phosphonovaleric acid (APV), had no effect on paired-pulse facilitation of either the LA or the PS. The gamma-aminobutyric acid-B (GABAB) agonist baclofen increased facilitation of the PS. This was mainly due to a reduction of the unconditioned response. Facilitation of the LA was reduced by both baclofen and the GABAB antagonist, 2-OH-saclofen. Baclofen increased the LA of the unconditioned response, while this was unaffected by 2-OH-saclofen. The LA of facilitated responses was decreased by 2-OH-saclofen while the effect of baclofen on these responses was more complex. Baclofen reduced the LA of maximally facilitated responses, while the LA of slightly facilitated responses was increased. The results show that different mechanisms are involved in the facilitation of the LA and the PS. Furthermore, activation of GABAB receptors makes a large contribution to paired-pulse facilitation of the field EPSP. It is also suggested that recording of extracellular fields in str. rad. in response to paired-pulse stimulation provides a simple electrophysiological model for testing the effect of agents which act at the GABAB receptor.

Topics: 2-Amino-5-phosphonovalerate; 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Baclofen; Electric Stimulation; Evoked Potentials; Female; Hippocampus; In Vitro Techniques; Male; Pyramidal Tracts; Quinoxalines; Rats; Receptors, GABA-A