NMDA receptor-mediated mechanism of ketamine-induced facilitation of glutamatergic excitatory synaptic transmission

The effect of ketamine on CA1-field EPSPs (fEPSPs) in rat hippocampal slices was investigated. Ketamine (100 microM) facilitated fEPSPs at 0.05 Hz. The fEPSP facilitation was suppressed completely by AP-5 and partially by propranolol, and also by an increase in stimulation frequency. These results indicate that ketamine facilitates excitatory synaptic transmission by activating NMDA receptors via beta-adrenoceptors under conditions in which NMDA receptor channel block is slight.     

Reduction of voltage-dependent currents by ethanol contributes to inhibition of NMDA receptor-mediated excitatory synaptic transmission

Previous studies have shown inhibitory effects of EtOH on NMDA receptor-mediated synaptic transmission in several brain regions. We examined this effect of EtOH under both current clamp and voltage clamp conditions in the basolateral amygdala because of the putative role of the amygdala in mediating anxiolytic effects of EtOH. We found that EtOH reduced NMDA receptor-mediated synaptic responses. In addition, we found that NMDA receptor-mediated depolarizations could also activate a voltage-dependent regenerative potential which was also sensitive to EtOH. Pharmacological characterization of this current was consistent with a high-threshold Ca²⁺ current. This current also exhibited a pronounced tendency towards transient enhancement upon withdrawal of EtOH.     

Adenosine inhibits excitatory but not inhibitory synaptic transmission in the hippocampus.

We examined the effects of adenosine and baclofen on inhibitory (IPSC) and excitatory (EPSC) synaptic currents in dissociated rat hippocampal neurons. Adenosine dramatically reduced monosynaptic EPSCs but failed to diminish IPSCs. This selective effect on EPSCs is likely due to inhibition of excitatory transmitter release because adenosine did not directly alter any properties of postsynaptic neurons. Baclofen depressed both EPSCs and IPSCs to approximately the same extent. These experiments indicate that the presynaptic effects of adenosine and baclofen are clearly separable and that transmitter sensitivities of inhibitory and excitatory neurons can differ. These differences could be exploited in the design of antiepileptic drugs that act at adenosine receptors to limit excitatory neurotransmission without blocking tonic inhibition.     

Interleukin-1beta abrogates long-term depression of hippocampal CA1 synaptic transmission

Although interleukin-1beta (IL-1beta) is well known to modulate synaptic transmission and plasticity of the hippocampus, no study has yet evaluated how this cytokine affects long-term depression (LTD), one of the major forms of hippocampal synaptic plasticity. Here we report that at Schaffer collateral-CA1 synapses, bath application of IL-1beta induces a long-lasting decrease in synaptic strength in intact slices, but not in disinhibited slices in the presence of bicuculline, a gamma-aminobutyric acid receptor antagonist. The IL-1beta-induced synaptic depression efficiently foreclosed the subsequent induction of LTD in response to a 1-Hz tetanus and, conversely, it was also prevented by preexisting LTD. These results suggest that IL-1beta-induced, persistent depression of synaptic efficacy is required for GABAergic activation and shares, at least in part, a common cellular mechanism for LTD.     

Development of NMDA and non-NMDA receptor-mediated excitatory synaptic transmission in geniculocortical and corticocortical connections in organotypic coculture preparations

Development of N-methyl-D-aspartate (NMDA) and non-NMDA receptor-mediated excitatory synaptic transmission was studied in the visual cortex using organotypic slice cocultures. A slice of visual cortex (VC) dissected from newborn rats was cocultured with either a chunk of embryonic lateral geniculate nucleus (LGN) or another VC. During 7-38 days in vitro (DIV), geniculocortical monosynaptic excitatory postsynaptic potentials (EPSPs) were recorded from layer IV neurons in response to stimulation of the LGN in cocultures of the VC with the LGN. Similarly, corticocortical monosynaptic EPSPs were recorded from layers II/III and V/VI neurons in cocultures of two VCs when stimulating the partner VC. The initial slopes of the non-NMDA and NMDA receptor-mediated components of the EPSPs, which were dissociated pharmacologically, were assessed and compared among three different culture stages, early (7-11 DIV), middle (12-15 DIV) and late (17-38 DIV). With progression of the culture stage, the non-NMDA component tended to increase in both the geniculocortical and corticocortical connections. In contrast, the NMDA component exhibited distinct developmental changes. The NMDA component in layer IV neurons, which receive geniculate inputs, showed a transient increase in the middle stage. In the corticocortical connection, the magnitude of the NMDA component was large in the early stage and maintained through all culture stages in layer V/VI cells, whereas in layer II/III cells it decreased sharply by the late stage. Our results suggest that glutamatergic transmission in the visual cortex develops differently in the geniculocortical and corticocortical connections.     

Baclofen selectively inhibits excitatory synaptic transmission in the hippocampus

The effect of baclofen was investigated on mossy fiber, Schaffer collateral and perforant path synaptic transmission in hippocampal slices. Baclofen completely inhibits mossy fiber and Schaffer collateral synaptic transmission with an IC50 of 3.8 microM. The lateral perforant path is insensitive to baclofen, while the response in the medial zone was partially blocked. Baclofen does not appear to act in a GABA-like manner.     

Fear conditioning enhances spontaneous AMPA receptor-mediated synaptic transmission in mouse hippocampal CA1 area

AMPA receptor-mediated synaptic modifications in the amygdala have been reported to sustain cued fear conditioning. However, the hippocampal formation is also critically involved in fear learning. Therefore, we examined whether fear conditioning is also accompanied by changes in AMPA receptor-mediated synaptic transmission in the hippocampus. We focused on spontaneous miniature excitatory post-synaptic currents (mEPSCs). Young adult mice were trained using tone/footshock pairings and contextual/cued memories were tested 3–4 h and 1 day later. We found that the mEPSC frequency was significantly enhanced when recorded 3 h, but not 24 h, after fear conditioning training. Fear training induced a slight enhancement in the mEPSC amplitude at 3 h after training. The increased mEPSC frequency and amplitude were absent in animals that were only exposed to footshock or novelty or unpaired tone/footshock training. This implies that learning the association between context, tone and footshock transiently enhances hippocampal CA1 spontaneous synaptic transmission, which may contribute to the encoding of the fearful event.     

Arylamine spider toxins antagonize NMDA receptor-mediated synaptic transmission in rat hippocampal slices.

The effects of arylamine spider toxins on synaptic transmission in rat hippocampal slices were investigated. Two different responses were monitored: the AMPA receptor-mediated population spike recorded in control buffer (selectively antagonized by DNQX) and the NMDA receptor-mediated EPSP recorded in nominally magnesium-free buffer containing 20 microM DNQX (selectively antagonized by AP5, AP7, and dizocilpine (MK-801)). The synthetic arylamine spider toxins JSTX-3, argiotoxin-636, and argiotoxin-659 were 26 to 73 times more potent at antagonizing the NMDA receptor-mediated EPSP (IC50 values ranging from 12 to 24 microM) than the AMPA receptor-mediated population spike (IC50 values ranging from 612 to 878 microM). These results indicate that arylamine spider toxins are selective antagonists of NMDA receptors in the mammalian CNS.     

Brain-derived neurotrophic factor enhances fast excitatory synaptic transmission in human epileptic dentate gyrus

Brain-derived neurotrophic factor (BDNF) has trophic effects and modulates synaptic transmission in the hippocampal formation in animal studies. It is also upregulated in acute and chronic epilepsy models and in human temporal lobe epilepsy. This study was undertaken to examine the effects of BDNF on fast synaptic transmission in the human epileptic dentate gyrus. Hippocampal specimens were acquired from patients with temporal lobe epilepsy during surgical removal of the anterior temporal lobe intended to treat the epileptic condition. Whole-cell patch-clamp recordings were obtained from dentate granule cells in transverse hippocampal slices in vitro. Application of BDNF increased the amplitude and frequency of spontaneous excitatory postsynaptic currents and increased the amplitude of evoked excitatory postsynaptic currents. BDNF had no effect on spontaneous inhibitory postsynaptic currents but produced a decrease in amplitude of evoked inhibitory postsynaptic currents. BDNF's effects were abolished by coapplication of the tyrosine kinase inhibitor K252a. Therefore, BDNF enhances fast excitatory transmission in the epileptic human dentate gyrus and may play an important role in epileptogenesis in temporal lobe epilepsy. This raises the possibility of designing therapies for this disorder that may be both anticonvulsant and antiepileptogenic.     

GABA(B) receptor-mediated heterosynaptic depression of excitatory synaptic transmission in rat frontal neocortex

Neocortical synapses display several forms of short-term plasticity including paired-pulse facilitation and depression. The mechanisms underlying this diversity are unclear. Synaptic currents in response to paired stimulation were recorded from layer II/III pyramidal neurons in rat frontal neocortical slices using the whole-cell patch-clamp method. Both paired-pulse facilitation (PPF) and paired-pulse depression (PPD) were observed in control saline. In the presence of 10 microM bicuculline (BIC), prominent PPD was consistently elicited. The maximal depression of the second EPSC occurred around 100 ms although PPD was still observed at intervals up to 1500 ms. Manipulations that reduced the probability of transmitter release significantly affected PPD. Both conditioning (C)- and test (T)-EPSCs were reduced when the extracellular Ca(2+) concentration was lowered from 3 to 1 mM. The decrease was greater in the C-EPSC resulting in a decrease in PPD. The gamma-aminobutyric acid (GABA)(B) receptor agonist baclofen (10 microM) reduced the amplitude of both evoked EPSCs and changed PPD to PPF. In the presence of the GABA(B) antagonists 2(OH)-saclofen (200-400 microM) or SCH50911 (10 microM), PPF was commonly observed. The metabotropic glutamate receptor antagonist MCPG (500 microM) had no effect on neocortical PPD. Brief stimulus trains induced a progressive depression that was insensitive to GABA(B) antagonists. Paired-pulse depression of excitatory synaptic transmission is a prominent phenomenon in frontal neocortex. At least two components of depression were observed. They may play an important role in regulating the balance between excitation and inhibition, therefore maintaining stability in cortical circuits.     

Ethanol inhibits epileptiform activity and NMDA receptor-mediated Synaptic transmission in rat amygdaloid slices

The effect of ethanol on the epileptiform activity induced by Mg(++)-free solution was studied in rat amygdalar slices using intracellular recording techniques. The spontaneous and evoked epileptiform discharges consisting of an initial burst followed by afterdischarges were observed 20-30 min after switching to Mg(++)-free medium. Superfusion with ethanol (20-100 mM) reversibly reduced the duration of spontaneous and evoked bursting discharges in a concentration-dependent manner. Synaptic response mediated by N-methyl-D-aspartate (NMDA) receptor activation was isolated by application of a solution containing the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and either in Mg(++)-free solution or in the presence of 50 microM bicuculline. Application of ethanol reversibly suppressed the duration of NMDA receptor-mediated synaptic response. These results suggest that intoxicating concentrations of ethanol possess anticonvulsant activity through blocking the NMDA receptor-mediated synaptic excitation. In addition, the observed effect of ethanol on NMDA receptor-mediated synaptic response could be relevant to the cognitive and behavioral manifestations seen in some alcoholics.     

Potentiation of NMDA receptor-mediated synaptic responses by microglia

To study the influence of microglia on glutamatergic synaptic transmission in the acute phase of neuronal injury, we first examined the effects of primary cultured microglia transferred onto the organotypic cortical slice cultures. In these microglia-transferred cortical slice cultures, stimulation of the subcortical white matter induced fast excitatory postsynaptic potentials followed by N-methyl-D-aspartate (NMDA) receptor-mediated plateau-like potentials that were never observed in control slice cultures. A similar potentiation of NMDA receptor-mediated postsynaptic responses was also observed by an application of a microglial-conditioned medium (MCM, 10% v/v) in acute cortical slices. These effects of MCM disappeared after boiling or incubation with proteinase K. After fractionation of MCM by anion-exchange chromatography, the enhancing activity of each fraction was quantitated electrophysiologically. When each fraction was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the fraction 24 which showed the most potent enhancing activity on NMDA receptor-mediated responses contained a relatively strong protein band with a molecular mass of approximately 70 kDa. MCM also enhanced both glutamate- and NMDA-induced inward currents recorded from acutely isolated cortical neurons. It was also noted that glutamate and NMDA induced transient large inward currents during an application of MCM, which were never observed in the control condition. These observations strongly suggest that NMDA receptor-mediated responses can be potentiated by both heat- and protease-labile (presumably 70-kDa proteins) molecules released from microglia.     

P2X receptor-mediated excitatory synaptic currents in somatosensory cortex

Fast P2X receptor-mediated excitatory postsynaptic current (EPSC) was found in pyramidal neurones of layer V of somatosensory cortex in slices acutely isolated from the brain of 17- to 22-day-old rats. The EPSCs were elicited by field electrical stimulation in the layer VI at 0.1 Hz in the presence of picrotoxin. When the glutamatergic EPSC was blocked by glutamate receptors inhibitors NBQX and D-AP5, a residual EPSC (rEPSC) was recorded from 85% of neurones tested. This rEPSC was not affected by blockers of nicotinic (hexamethonium) and serotonin (Y25130) receptors; however, it was reversibly inhibited by P2X receptors antagonists (NF023, NF279, and PPADS). An application of ATP (20 microM), beta,gamma-methylene ATP (25 microM), and alpha,beta-methylene ATP (20 microM) to acutely isolated pyramidal neurones of layer V evoked inward currents (30 to 400 pA) in 75% of cells tested. We concluded that several subtypes of P2X purinoreceptors participate in synaptic transmission in neocortex.     

Homer-1a/Vesl-1S enhances hippocampal synaptic transmission

Homer/Vesl proteins are involved in regulating metabotropic glutamate receptors, synaptogenesis, dendritic spine development and axonal pathfinding. We investigated the potential modulation of glutamatergic synaptic transmission by the immediate early gene product Homer-1a/Vesl-1S and by the constitutively expressed long-form Homer-1c/Vesl-1L in CA1 pyramidal cells from cultured rat hippocampal slices. Semliki Forest virus vector-mediated overexpression of Homer-1a enhanced alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor function, but did not detectably affect N-methyl-d-aspartate (NMDA) receptor function and presynaptic glutamate release. Overexpression of Homer-1c, by contrast, did not alter synaptic transmission. To corroborate our electrophysiological results obtained in slice cultures, we performed quantitative immunocytochemistry in cultures of dissociated hippocampal neurons. Homer-1a also increased synaptic clustering of AMPA but not NMDA receptors, whereas Homer-1c had no detectable effect. Our results show that Homer-1a potentiates synaptic AMPA receptor function, supporting a critical role for Homer-1a in hippocampal synaptic plasticity.     

Topiramate alters excitatory synaptic transmission in mouse hippocampus

Antiepileptic drugs may exert neuroprotective effects by decreasing excessive membrane excitability, neurotransmitter release, or postsynaptic Ca2+ entry. To assess these sites of action, we combined fluorescence Ca2+ imaging with extracellular field recording to analyze axonal excitability, evoked presynaptic Ca2+ entry through presynaptic Ca2+ channels, postsynaptic excitatory field potentials (fEPSP), and postsynaptic Ca2+ buildup ([Capost]) at the mouse hippocampal CA3-CA1 synapse exposed to topiramate (TPM). Topiramate had no effect on presynaptic Ca2+ entry, and produced only a minor inhibition of axonal excitability. Topiramate at concentrations up to 100 microM only slightly reduced the amplitude of the evoked fEPSP, but strongly inhibited the [Capost] evoked by repetitive synaptic activation. Postsynaptically, the action of TPM on the fEPSP and [Capost] was not mediated by an inhibition of the NMDA receptor, or by direct modulation of voltage-dependent Ca2+ channels, but reflected reduced somatic or dendritic membrane depolarization by AMPA and kainate receptors. These results are consistent with the known anticonvulsant properties of TPM. In addition, the ability of TPM to reduce postsynaptic Ca2+ buildup may provide a potential mechanism for neuronal protection during paroxysmal firing associated with epileptic seizures.     

Protein kinase Mzeta enhances excitatory synaptic transmission by increasing the number of active postsynaptic AMPA receptors

Protein kinase Mzeta (PKMzeta), a constitutively active, atypical PKC isoform, enhances synaptic strength during the maintenance of long-term potentiation (LTP). Here we examine the mechanism by which PKMzeta increases synaptic transmission. Postsynaptic perfusion of PKMzeta during whole-cell recordings of CA1 pyramidal cells strongly potentiated the amplitude of AMPA receptor (AMPAR)-mediated miniature EPSCs (mEPSCs). Nonstationary fluctuation analysis of events recorded before and after PKMzeta enhancement showed that the kinase doubled the number of functional postsynaptic AMPAR channels. After sustained potentiation, application of a PKMzeta inhibitor reversed the increase in functional channel number to basal levels, suggesting that persistent increase of PKMzeta is required to maintain the postsynaptic localization of a mobile subpopulation of receptors. The kinase did not affect other sites of LTP expression, including presynaptic transmitter release, silent synapse conversion, or AMPAR unit conductance. Thus PKMzeta functions specifically to establish and maintain long-term increases in active postsynaptic AMPAR number.     

Lipocortin-1 inhibits NMDA receptor-mediated neuronal damage in the striatum of the rat

Lipocortin-1 (annexin-1), an endogenous phospholipid and calcium binding protein, has been shown to significantly attenuate the damage produced by focal cerebral ischaemia in the rat. In the present study we have therefore investigated its effect on N-methyl-D-aspartate (NMDA) induced neuronal damage. Unilateral intrastriatal infusion of a potent and selective NMDA agonist, cis-2,4-methanoglutamate (MGlu), induced an extensive lesion of the striatum in the rat, which was inhibited (greater than 80%) by prior injection of MK801 (4 mg/kg, i.p.). Infusion of 1.2 micrograms of an active fragment of lipocortin-1 (N-terminal 1-188 aa) immediately after MGlu significantly reduced the extent of damage by 44.2 +/- 8.0%. In contrast, infusion of 3 microliters of neutralizing anti-lipocortin-1 antibody with MGlu increased lesion size by 158.9 +/- 22.0%. These findings indicate that the damage produced by intrastriatal infusion of MGlu is mediated by the NMDA receptor. Lipocortin-1 fragment markedly attenuated, and the neutralizing antibody increased, this NMDA mediated neuronal damage. These observations may explain the neuroprotective action of lipocortin following cerebral ischaemia.