Extracts from rhizomes of Cyperus articulatus (Cyperaceae) displace [3H]CGP39653 and [3H]glycine binding from cortical membranes and selectively inhibit NMDA receptor-mediated neurotransmission.

The marshland plant Cyperus articulatus (Cyperaceae) is commonly used in traditional medicine in Africa and Latin America to treat a wide variety of human diseases ranging from headache to epilepsy. We tested the hypothesis that the purported anti-epileptic effect of this plant might be due to a functional inhibition of excitatory amino acid receptors. One or several component(s) contained in the extracts inhibited the binding of [3H]CGP39653 to the NMDA recognition site and of [3H]glycine to the strychnine-insensitive glycine site of the NMDA receptor complex from rat neocortex. Water extracts from rhizomes of Cyperus articulatus dose-dependently reduced spontaneous epileptiform discharges and NMDA-induced depolarizations in the rat cortical wedge preparation at concentrations at which AMPA-induced depolarizations were not affected. We conclude that the purported beneficial effects of Cyperus articulatus might at least partially be due to inhibition of NMDA-mediated neurotransmission.

Spontaneous and stimulation-induced synchronized burst afterdischarges in the isolated CA1 of kainate-treated rats.

1. Kainate treatment preferentially kills dentate hilar neurons and CA3 pyramidal cells and ultimately leads to a chronic epileptic state. Bicuculline-induced epileptiform bursts were studied to test the hypothesis that multiple kainate injections and consequent status epilepticus would lead-after weeks to months of recovery-to prolonged synchronous afterdischarges in the isolated CA1 area of rat hippocampal slices, as would be expected if new recurrent excitatory circuits had formed. 2. Synaptic responses evoked in CA1 pyramidal cells of rats injected subcutaneously with kainate (10 hourly injections, 5 mg/kg each) 24-316 days before the slice experiment were compared with responses in slices from untreated and saline-injected controls. The maximal response to stratum radiatum stimulation in normal solution consisted of two to eight population spikes. 3. When gamma-aminobutyric acid-A receptor-mediated inhibition was reduced with bicuculline, synchronized burst afterdischarges after the initial stimulation-evoked burst, similar to the type of activity described in area CA3 under conditions where inhibition is impaired, occurred in 23% of slices. 4. The prolonged synchronized burst afterdischarges in the isolated CA1 area of kainate-treated rats were associated with large excitatory postsynaptic potentials (EPSPs). These prolonged bursts were not graded with the stimulus intensity; rather, they were triggered in an all-or-none manner, even though there was some variability across bursts. The bursts of population spikes also were correlated with subthreshold EPSPs. 5. Slices that had synchronized burst afterdischarges had significantly more damage in area CA3 than slices without afterdischarges. 6. The data indicate that kainate-induced damage in CA3 can lead to prolonged synchronous afterdischarges, even after CA1 is surgically isolated from the CA3 area. Because the repetitive bursts during the prolonged and synchronous afterdischarges were associated with large EPSPs, these data suggest that kainate-induced damage to CA3 and subsequent degeneration of synaptic terminals in the CA1 area causes the formation of new recurrent excitatory circuits that could be involved in the development of chronic epilepsy.

Biphenyl-derivatives of 2-amino-7-phosphonoheptanoic acid, a novel class of potent competitive N-methyl-D-aspartate receptor antagonist--I. Pharmacological characterization in vitro.

Omega-Phosphono-substituted alpha-amino acids have long been known to be antagonists at the N-methyl-D-aspartate (NMDA) receptor. D-2-Amino-5-phosphonopentanoic (D-AP5) and D-2-amino-7-phosphonoheptanoic (D-AP7) acids are the "prototype" compounds of this kind. Insertion of a biphenyl-moiety in the middle of the AP7 chain results in increased affinity and reverses the enantioselectivity from a D- to an L-form preference (Müller et al., (1992) Helv. Chim. Acta 75: 855-864). We describe here a series of substituted biphenyl-AP7-derivatives, the most potent of which have affinities (in a [3H]CGP-39653 binding assay using native and recombinant receptors) and potencies (antagonism of NMDA-induced depolarizations in a cortical wedge preparation; inhibition of glutamate-stimulated [3H]MK-801 binding under non-equilibrium conditions) in the low nanomolar range. Structure-activity relationships show that hydroxy-substitution at the C5-atom in the AP7-chain as well as substitution in the second phenyl ring with space filling (such as chloro-)groups in the para- and especially the ortho-position (extending the torsion angle of the two rings) increase affinity and potency of these compounds. They have no relevant affinities for the strychnine-insensitive glycine co-agonist site or the MK-801/PCP channel blocking site on the NMDA receptor complex. AMPA- and kainate-induced responses were not affected by biphenyl-analogues. These compounds also do not interact with a number of other neurotransmitter receptor sites, and they do not inhibit the uptake of [3H] glutamate in rat brain synaptosomes. However, they display affinities in the (sub)micromolar range for a non-NMDA, non-AMPA, non-kainate binding site for [3H]glutamate, measured in the presence of calcium chloride, the functional correlate of which has not yet been elucidated.

Thalamocortical and corticocortical excitatory postsynaptic potentials mediated by excitatory amino acid receptors in the cat motor cortex in vivo.

Intracellular recordings were made from neurons in the motor cortex of an anaesthetized cat, together with iontophoretic application of excitatory amino acid receptor agonists and antagonists, in order to evaluate the role of such receptors in excitatory postsynaptic potentials evoked from stimulation of afferent and recurrent pathways in vivo. Excitatory postsynaptic potentials which were evoked by stimulation of the ventrolateral thalamus were found to be largely insensitive to antagonism by N-methyl-D-aspartate receptor antagonists, although they were susceptible to blockade by the non-N-methyl-D-aspartate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione. Increasing the ventrolateral thalamus stimulation frequency from 0.5 or 1 to 5 Hz caused an increase of evoked excitatory postsynaptic potential amplitudes and number of action potentials. These augmented excitatory postsynaptic potentials remained insensitive to application of N-methyl-D-aspartate antagonists. In contrast, recurrent excitatory postsynaptic potentials evoked by stimulation of the pyramidal tract were found to be sensitive to N-methyl-D-aspartate receptor antagonists and/or non-N-methyl-D-aspartate receptor antagonists in some neurons. These results demonstrate the involvement of both N-methyl-D-aspartate- and non-N-methyl-D-aspartate receptors in synaptic responses of cat motor cortex neurons in vivo, and that the synaptic pharmacology of the thalamic input may differ from that of the local recurrent pathways.

N-methyl-D-aspartate induces regular firing patterns in the cat lateral habenula in vivo.

The present study was undertaken to elucidate the action of excitatory amino acids in the dorsal diencephalic pathway. Single neurons in the lateral habenula of halothane-anesthetized cats were recorded extracellularly, and excitatory amino acid receptor agonists and antagonists were applied by iontophoresis. Most neurons in the lateral habenula were spontaneously active. This spontaneous firing could be inhibited by kynurenic acid, a broad spectrum antagonist of excitatory amino acid receptors, but not by the selective N-methyl-D-aspartate receptor antagonist 2-amino-7-phosphono-heptanoic acid. Iontophoretic application of alpha-amino-3-hydroxy-5-methyl-5-isoxazolepropionate, quisqualate and kainate mostly elicited a non-burst, regular firing pattern which was sensitive to kynurenic acid. Surprisingly, 116 (96%) out of 121 neurons in the lateral habenula responded to iontophoretic application of N-methyl-D-aspartate with a regular non-burst firing pattern, in contrast to previously published observations from other brain regions where N-methyl-D-aspartate predominantly elicited phasic firing patterns. When cells were recorded with electrode assemblies where one iontophoretic barrel contained MgCl2 or MgSO4, only 10 (43%) out of 23 cells responded with regular firing upon application of N-methyl-D-aspartate, while 13 (57%) now displayed a phasic firing pattern. In these cells iontophoretically applied alpha-amino-3-hydroxy-5-methyl-5-isoxazolepropionate or quisqualate still evoked only regular firing. In a few cases, an initially regular N-methyl-D-aspartate-induced firing pattern could be changed to a phasic pattern following active ejection of Mg2+ ions.(ABSTRACT TRUNCATED AT 250 WORDS)

Persistent hyperexcitability in isolated hippocampal CA1 of kainate-lesioned rats.

1. Subcutaneous kainate injection in rats evoked acute seizures and led to cell loss in the hilus and areas CA1 and CA3, which resembled the pattern of hippocampal sclerosis often associated with temporal lobe epilepsy in humans. 2. Simultaneous intra- and extracellular recordings were performed in the stratum pyramidale of area CA1 while stimulating in the stratum radiatum close to the recording electrodes. Responses from control slices consisted of a brief excitatory postsynaptic potential (EPSP) with only one action potential, corresponding to a single extracellular population spike, followed by a clear biphasic inhibitory postsynaptic potential (IPSP). In slices from kainate-treated animals, however, stimulation evoked a prolonged EPSP, which often triggered multiple action potentials corresponding to multiple extracellular population spikes. 3. In slices from kainate-treated animals, the mean amplitude but not the duration of the stimulation-evoked IPSP was reduced. The extent of the kainate-induced loss of inhibition in area CA1 was highly variable. 4. Low concentrations of bicuculline in control slices led to a moderate hyperexcitability, which consisted of multiple population spikes and mirrored the responses observed in slices from kainate-treated animals in normal ACSF. Prolonged application of 10-30 microM bicuculline for > or = 30 min led to a much higher level of hyperexcitability, which was similar in slices from controls and kainate-treated rats. These findings are consistent with the hypothesis that the hyperexcitability of CA1 pyramidal neurons following kainate treatment is mainly due to decreased GABAA-receptor-mediated inhibition and that the loss of inhibition is only partial.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential effects of (D)- and (L)-homocysteic acid on the membrane potential of cat caudate neurons in situ.

The enantiomers of homocysteic acid have been applied by microiontophoresis to neurons of the cat caudate nucleus in situ. The (L)-enantiomer elicited a bursty firing pattern similar to the one caused by N-methyl-D-aspartate, but differing from the N-methyl-D-aspartate pattern inasmuch as (L)-homocysteate induced depolarization shifts were shorter and had a smaller amplitude. (L)-Homocysteate induced excitations could be strongly inhibited by the selective N-methyl-D-aspartate antagonist 2-amino-7-phosphonoheptanoic acid but they were less sensitive to this antagonist than N-methyl-D-aspartate itself. (D)-Homocysteate elicited a more regular firing pattern similar to the one caused by non-N-methyl-D-aspartate excitatory amino acids such as quisqualate. These excitations were only rarely inhibited by 2-amino-7-phosphonoheptanoic acid. Our results suggest that (L)-homocysteate, a transmitter candidate at central mammalian synapses, is a mixed excitatory amino acid agonist with a strong preference for N-methyl-D-aspartate receptors in the cat caudate nucleus, while (D)-homocysteate has a predominant action at non-N-methyl-D-aspartate excitatory amino acid receptors.