Age-related behavioral, morphological and physiological changes in the hippocampus of zitter rats.

Convulsive seizure is known to be associated with hippocampal abnormalities, such as hilar cell degeneration, abnormal mossy fiber sprouting in the dentate gyrus (DG) and abnormal expression of immediate early genes. However, whether these morphological changes are a cause or consequence of convulsive seizures has remained contentious. Zitter (zi/zi) rats carry a mutation of the attractin gene and display spongiform degeneration of the brain. Spontaneous convulsive seizures in zi/zi rats over 8 months (M) old were demonstrated using 24-h video monitoring. Spontaneous convulsive seizures did not occur before this age. The present study examined structural changes in the hippocampus of zi/zi rats at different ages. Fluoro-Jade B-positive cells first appeared in the hilus of 1-M zi/zi rats, indicating hilar cell degeneration. After 2 M, mossy fiber sprouting was observed in granular cell layers and in the inner molecular layer. After 10 M, granule cells showed Fos expression. In the hippocampal slices from 12-M zi/zi rats, abnormal population spikes in the DG were observed in the presence of bicuculline and strychnine. Conversely, Sprague-Dawley rats showed no aberrant zinc distribution, few Fos-positive cells, no Fluoro-Jade B-positive cells in the hippocampus and no abnormal population spikes in the DG. These data indicate that morphological changes in the hippocampus might contribute to epileptogenesis in this mutant rat.

Peripheral nerve injury-induced rearrangement of neural circuit in the spinal dorsal horn revealed by cross-correlation analysis.

Peripheral nerve injury often induces abnormal pain states, such as hyperalgesia and allodynia. In this study, we attempted to elucidate how neurons are synaptically integrated into the neuronal circuitry in the spinal dorsal horn and how synaptic connectivity patterns among dorsal horn neurons are altered by peripheral nerve injury. Experiments were performed on 6-8-week-old ICR mice. Partial sciatic nerve ligation was performed. Transverse slices of the lumbar spinal cord were prepared. Spike activities were simultaneously recorded from multiple neurons in the superficial dorsal horn (SDH) using a multi-electrode array system, and cross-correlograms between spike trains of neuron pairs were constructed. In sham-operated control mice, except for the flat cross-correlogram, the most common pattern was a cross-correlogram suggestive of common excitatory synaptic inputs to neuronal pairs. Peripheral nerve ligation increased the incidences of cross-correlograms suggestive of common excitatory synaptic inputs and excitatory synaptic connections, and decreased that of inhibitory synaptic connections. Additionally, bath-applied capsaicin, an agonist for transient receptor potential vanilloid 1 receptor, increased the frequency of action potentials. The effects of capsaicin stimulation on the incidence of cross-correlograms with various patterns were significantly different between sham-operated control and sciatic nerve-ligated mice. The present observations seem to indicate that neurons in the SDH form excitatory and/or inhibitory synapses with nearby neurons, and that synaptic connections among neurons in the SDH may remarkably change after the development of neuropathic pain.

Effects of pregabalin on spinal d-serine content and NMDA receptor-mediated synaptic transmission in mice with neuropathic pain.

Pregabalin (PGB) is a chemical derivative of the inhibitory neurotransmitter γ-aminobutyric acid, and is successfully used for the treatment of neuropathic pain. Substantial evidence suggests that d-serine, an endogenous co-agonist at the strychnine-insensitive glycine site of the NMDA receptor, counteracts the antinociceptive actions of PGB at the level of the spinal cord. In the present study, we examined the impact of PGB treatment on spinal d-serine content and NMDA receptor-mediated synaptic transmission in the superficial dorsal horn of peripheral nerve-ligated neuropathic mice. Mechanical allodynia was assessed using von Frey filaments. On post-surgical day 9 (after 5days of treatment with PGB [50mg/kg] or saline vehicle), the lumbar spinal cord was removed, homogenized, and ultrafiltrated. Supernatant samples were treated with Marfey's reagent and analyzed with liquid chromatography-mass spectrometry to measure d-serine content. In the electrophysiological experiments, tight-seal whole-cell recording was performed on neurons in the superficial dorsal horn of spinal cord slices. Partial sciatic nerve ligation increased spinal d-serine content, increased the NMDA/non-NMDA ratio of EPSC amplitudes, and slowed the decay phase of the NMDA component of EPSCs (NMDA-EPSCs). PGB treatment attenuated mechanical allodynia and reduced spinal d-serine content, decreased the NMDA/non-NMDA ratio, and shortened the decay time of NMDA-EPSCs. Furthermore, bath-applied d-serine attenuated the effects of PGB treatment. Although the precise mechanism for the effect of PGB on d-serine metabolism and abundance is unknown, the antinociceptive action of PGB likely involves the reduction of spinal d-serine content and subsequent attenuation of NMDA receptor-mediated synaptic transmission in the superficial dorsal horn.

Clinical utility of automated ablation lesion tagging based on catheter stability information (VisiTag Module of the CARTO 3 System) with contact force-time integral during pulmonary vein isolation for atrial fibrillation.

BACKGROUND: The clinical utility of an automated lesion tagging module based on catheter stability information (VisiTag) with the CARTO system during atrial fibrillation (AF) ablation remains to be established. We investigated whether VisiTag-guided extensive encircling pulmonary vein isolation (EEPVI) produces durable lesions. METHODS: The study involved 54 patients undergoing EEPVI for paroxysmal AF. We performed EEPVI guided by the module-generated ablation tags, i.e., "VisiTags," which are point-by-point ablation tags placed on 3D maps. The patients were divided into two groups: those treated under a moderate catheter stability VisiTag setting, i.e., a 3-mm distance limit for at least 5 s and a minimum contact force (CF) of 8 g over 25 % of the set time period with a target force-time integral (FTI) ≥300 g*s (n = 27), and those treated under a strict catheter stability setting, i.e., a 3-mm distance limit for at least 10 s and a minimum CF of 10 g over 50 % of the set time period with a target FTI ≥400 g*s (n = 27). RESULTS: After EEPVI, adenosine triphosphate-provoked dormant PV conduction was observed in six (22 %) patients in the moderate catheter stability group and in one (4 %) patient in the strict catheter stability group (p = 0.1003); the 12.9-month success rate was 81 % in both groups. CONCLUSIONS: The strict catheter stability setting for automated lesion tagging together with a target FTI of >400 g*s, vs. the moderate catheter stability setting with a target FTI of >300 g*s, produces less frequent ATP-provoked PV conduction and yields a comparably high mid-term success rate.

Presynaptic inhibitory actions of pregabalin on excitatory transmission in superficial dorsal horn of mouse spinal cord: further characterization of presynaptic mechanisms.

Pregabalin is widely used as an analgesic for the treatment of neuropathic pain. In the present experiments using mouse spinal slices, we recorded electrically evoked glutamatergic excitatory postsynaptic currents (eEPSCs) from superficial dorsal horn neurons. Pregabalin reduced the amplitude of eEPSCs, and increased the paired pulse ratio. Pregabalin also inhibited the frequency of spontaneously occurring miniature EPSCs without affecting their amplitude. Partial ligation of the sciatic nerve increased the expression of the calcium channel α2δ-1 subunit, and increased the presynaptic inhibitory action of pregabalin. Intrathecal injection of antisense oligodeoxynucleotide against the α2δ-1 subunit, decreased the expression of α2δ-1 mRNA in the spinal dorsal horn, and decreased pregabalin's action. These results provide further evidence that pregabalin exerts its presynaptic inhibitory action via binding with the α2δ subunit in a state-dependent manner. Furthermore, presynaptic actions of pregabalin were attenuated in knockout mice lacking the protein syntaxin 1A, a component of the synaptic vesicle release machinery, indicating that syntaxin 1A is required for pregabalin to exert its full presynaptic inhibitory action. These observations might suggest that direct and/or indirect interactions with the presynaptic proteins composing the release machinery underlie at least some part of pregabalin's presynaptic actions.

Differential expression of NMDA receptors in serotonergic and/or GABAergic neurons in the midbrain periaqueductal gray of the mouse.

N-methyl-d-aspartate (NMDA) receptors expressed in the midbrain periaqueductal gray (PAG) exert various physiological functions. The PAG contains various neurotransmitter phenotypes, which include GABAergic neurons and serotonergic neurons. In the present experiments, we made tight-seal whole-cell recordings from GABAergic and/or serotonergic neurons in mouse PAG slices and analyzed NMDA and non-NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation. The NMDA/non-NMDA ratio of EPSC amplitude was high and the decay time course of NMDA-EPSC was slow in non-serotonergic/GABAergic neurons. In contrast, serotonergic neurons exhibited a low NMDA/non-NMDA ratio and a fast decay time course of NMDA-EPSC. Peripheral nerve ligation-induced chronic pain was associated with an increased NMDA/non-NMDA ratio in serotonergic neurons. Additionally, single-cell real-time RT-PCR analysis showed that peripheral nerve ligation up-regulated NR2B subunit expression in non-serotonergic/non-GABAergic neurons. Such changes in NMDA receptor expression in the PAG result in an alteration of the descending modulation of nociception, which might be an underlying mechanism for peripheral nerve injury-evoked persistent pain. Finally, the expression of NMDA receptors seems differentially regulated among neurons of different neurotransmitter phenotypes in the PAG.

Functional roles of endogenous D-serine in pain-induced ultrasonic vocalization.

The N-methyl-D-aspartate receptor (NMDAR) is crucial for pain-related behaviors. D-Serine is synthesized from L-serine by serine racemase (SR) and modulates NMDAR functions by acting as an agonist at the glycine-binding site. We analyzed noxious stimulus-induced ultrasonic vocalization and locomotor activity in the open-field test using SR knockout (SR-KO) mice to examine the role of endogenous D-serine in mammalian behaviors. SR-KO mice emitted less ultrasonic vocalization after noxious stimulation (VAS) than wild-type (WT) mice. The locomotor activity of WT mice decreased with repeated daily exposures to the open field, whereas that of SR-KO mice remained unchanged. VAS was significantly enhanced during arthritis in WT mice, whereas it was not enhanced during arthritis in SR-KO mice. These results indicate that mice lacking the ability to produce D-serine endogenously in the brain differ from normal mice with respect to the chronic pain-induced behavioral changes.

Functional roles of endogenous D-serine in the chronic pain-induced plasticity of NMDAR-mediated synaptic transmission in the central amygdala of mice.

The amygdala is implicated in chronic pain-induced emotional changes. Chronic pain induces plastic changes of the N-methyl-d-aspartate receptor (NMDAR) functions in the brain including the amygdala. d-Serine is synthesized endogenously by serine racemase and modulates NMDAR-mediated synaptic transmission as a coagonist of glycine binding site. To clarify the functional roles of endogenous d-serine in chronic pain-induced plasticity of NMDAR mediated synaptic transmission, we investigated the NMDAR-mediated excitatory synaptic current (EPSC) of neurons in the latero-capsular division of the central amygdala (CeLC) using brain slices from serine racemase knockout (SR-KO) mice with chronic pain induced by monoarthritis. The decay time of NMDAR-mediated EPSC was significantly elongated by monoarthritis in wild type (WT) mice, but not in SR-KO mice. The d-serine application-induced increase of NMDAR-mediated EPSC was significantly facilitated by monoarthritis in WT mice, but not in SR-KO mice. These results suggest that endogenous d-serine facilitates chronic pain-induced plastic changes of NMDAR mediated synaptic transmission in CeLC.

Physiological properties of enkephalin-containing neurons in the spinal dorsal horn visualized by expression of green fluorescent protein in BAC transgenic mice.

BACKGROUND: Enkephalins are endogenous opiates that are assumed to modulate nociceptive information by mediating synaptic transmission in the central nervous system, including the spinal dorsal horn. RESULTS: To develop a new tool for the identification of in vitro enkephalinergic neurons and to analyze enkephalin promoter activity, we generated transgenic mice for a bacterial artificial chromosome (BAC). Enkephalinergic neurons from these mice expressed enhanced green fluorescent protein (eGFP) under the control of the preproenkephalin (PPE) gene (penk1) promoter. eGFP-positive neurons were distributed throughout the gray matter of the spinal cord, and were primarily observed in laminae I-II and V-VII, in a pattern similar to the distribution pattern of enkephalin-containing neurons. Double immunostaining analysis using anti-enkephalin and anti-eGFP antibodies showed that all eGFP-expressing neurons contained enkephalin. Incubation in the presence of forskolin, an activator of adenylate cyclase, increased the number of eGFP-positive neurons. These results indicate that eGFP expression is controlled by the penk1 promoter, which contains cyclic AMP-responsive elements. Sections obtained from sciatic nerve-ligated mice exhibited increased eGFP-positive neurons on the ipsilateral (nerve-ligated side) compared with the contralateral (non-ligated side). These data indicate that PPE expression is affected by peripheral nerve injury. Additionally, single-neuron RT-PCR analysis showed that several eGFP positive-neurons in laminae I-II expressed glutamate decarboxylase 67 mRNA and that some expressed serotonin type 3 receptors. CONCLUSIONS: These results suggest that eGFP-positive neurons in laminae I-II coexpress enkephalin and γ-aminobutyric acid (GABA), and are activated by forskolin and in conditions of nerve injury. The penk1-eGFP BAC transgenic mouse contributes to the further characterization of enkephalinergic neurons in the transmission and modulation of nociceptive information.

Facilitatory actions of serotonin type 3 receptors on GABAergic inhibitory synaptic transmission in the spinal superficial dorsal horn.

Analgesic effects of serotonin (5-hydroxytryptamine [5-HT]) type 3 (5-HT3) receptors may involve the release of gamma-aminobutyric acid (GABA) in the spinal dorsal horn. However, the precise synaptic mechanisms for 5-HT3 receptor-mediated spinal analgesia are not clear. In this study, we investigated whether GABAergic neurons in the superficial dorsal horn (SDH) express functional 5-HT3 receptors and how these 5-HT3 receptors affect GABAergic inhibitory synaptic transmission in the SDH, by using slice preparations from adult glutamate decarboxylase 67-green fluorescent protein (GAD67-GFP) knock-in mice. Tight-seal whole cell recordings from GFP-positive and -negative neurons showed that 5-HT3 receptor-specific agonist 2-methyl-serotonin (2-Me-5-HT) induced inward currents in a substantial population of both GFP-positive and -negative neurons. Additionally, we confirmed expression of 5-HT3 receptors in both types of neurons by single-cell reverse transcription-polymerase chain reaction (RT-PCR) analysis. Further, GABAA receptor-mediated inhibitory postsynaptic currents (IPSCs)-both those evoked by electrical stimulation and those occurring spontaneously in tetrodotoxin (i.e., miniature IPSCs [mIPSCs])-were recorded from GFP-negative neurons. 2-Me-5-HT increased the amplitude of the evoked IPSCs and the frequency of mIPSCs. The amplitude of mIPSCs was not affected by 2-Me-5-HT, suggesting that 5-HT augments GABAergic synaptic transmission via presynaptic mechanisms. The present observations indicate that 5-HT3 receptors are expressed on both somadendritic regions and presynaptic terminals of GABAergic neurons and regulate GABAA receptor-mediated inhibitory synaptic transmission in the SDH. Taken together, these results provide clues for the underlying mechanisms of the antinociceptive actions of 5-HT3 receptors in the spinal dorsal horn.

Presynaptic large-conductance calcium-activated potassium channels control synaptic transmission in the superficial dorsal horn of the mouse.

Large-conductance calcium-activated potassium channels (BK channels) have been suggested to play a substantial role in synaptic transmission in the spinal cord dorsal horn. In the present experiments, we attempted to clarify the physiological significance of BK channels in the modulation of synaptic transmission in the superficial dorsal horn where nociceptive information is processed. Spontaneously occurring excitatory postsynaptic currents (sEPSCs) were recorded from the neurons located in the superficial dorsal horn of a mouse spinal cord slice, and the effects of iberiotoxin, a BK channel blocker, on sEPSCs were analyzed. The frequency of sEPSCs was significantly higher in the peripheral nerve-ligated neuropathic mice than in the sham-operated control mice, but the amplitude of sEPSCs was equivalent between the two groups. Iberiotoxin increased the frequency of sEPSCs in the control mice to the same level as that in the neuropathic mice without affecting the amplitude of sEPSCs. In contrast, iberiotoxin did not show any significant effects on the sEPSCs in the neuropathic mice. These findings suggest that the BK channels that are located in presynaptic terminals control synaptic transmission in the superficial dorsal horn, and that functional downregulation of BK channels accompanies the neuropathic pain induced by peripheral nerve injury. This downregulation was confirmed by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis of the BK channel alpha subunit. Taken together, our present results indicate that BK channels play crucial roles in the synaptic transmission of nociceptive information in the superficial dorsal horn.

Facilitatory effects of subanesthetic sevoflurane on excitatory synaptic transmission and synaptic plasticity in the mouse hippocampal CA1 area.

Behavioral investigations have shown that general anesthetics at low concentration have enhancing effects on learning and memory in some animal models. In the present experiments, in order to elucidate the cellular mechanisms underlying such memory enhancement, the effects of anesthetics at low doses on synaptic plasticity in the hippocampus were investigated. Tight-seal whole-cell recordings were made from CA1 pyramidal cells in hippocampal slices prepared from adult male mice, and the effects of subanesthetic concentrations of the volatile anesthetic sevoflurane on the glutamatergic excitatory postsynaptic currents (EPSCs) were investigated. In addition, extracellular recordings of field excitatory postsynaptic potential (fEPSP) and population spike (PS) were made, and the effects of subanesthetic sevoflurane on long-term potentiation (LTP) of the fEPSP slope and on LTP of PS amplitude were analyzed. Sevoflurane at anesthetic concentration inhibited the amplitude of EPSCs with an increase in the paired-pulse facilitation (PPF) ratio. In contrast, subanesthetic sevoflurane increased the amplitude of EPSCs without any appreciable changes in the PPF ratio. Subanesthetic sevoflurane also showed facilitatory influences on LTP of PS amplitude but not on LTP of the fEPSP slope. These observations suggest that sevoflurane at anesthetic concentration presynaptically inhibits excitatory synaptic transmission and at subanesthetic concentration postsynaptically enhances excitatory synaptic transmission in the hippocampal CA1 region. Further, subanesthetic sevoflurane seems to exert facilitatory effects on the EPSP-to-spike coupling process in the postsynaptic neurons. These results might provide clues as to the cellular mechanism of light level of sevoflurane anesthesia.

Syntaxin 1A occludes GABA(B) receptor-induced inhibition of exocytosis downstream of Ca(2+) entry in mouse hippocampal neurons.

The mechanisms underlying gamma-amino butyric acid (GABA(B)) receptor-mediated inhibition of exocytosis have been characterized in a variety of synapses. Using patch-clamp recording methods, we attempted to clarify the intracellular mechanisms underlying presynaptic inhibition in autaptic synapses of isolated mouse hippocampal neurons. Baclofen, a selective GABA(B) receptor agonist, decreased the frequency of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) without changing their amplitude in Ca(2+)-free extracellular solution, suggesting that baclofen inhibits exocytosis downstream of Ca(2+) entry. Syntaxin 1A is known to modulate exocytosis and suppress neuronal sprouting. Antisense oligonucleotide-mediated knockdown of syntaxin 1A increased the frequency of mEPSCs under Ca(2+)-free condition. Estimation of the number of functional release sites by staining with FM1-43 indicated that the increased frequency of mEPSCs was induced by facilitation of exocytosis at each site, rather than by an increased number of release sites due to neuronal sprouting. Baclofen reduced mEPSC frequency in syntaxin 1A-knockdown neurons to the same level as that in nonsense oligonucleotide transfected neurons under Ca(2+)-free condition. These results suggest that the GABA(B) receptor- and syntaxin 1A-induced inhibitions of exocytosis occlude one another and that the GABA(B) receptor shares a common intracellular pathway with syntaxin 1A in inhibiting transmitter release downstream of Ca(2+) entry.

NMDA receptor 2B subunit-mediated synaptic transmission in the superficial dorsal horn of peripheral nerve-injured neuropathic mice.

Previous research has shown that peripheral inflammation and peripheral nerve injury alter the properties of NMDA receptors in the spinal dorsal horn. However, there is no direct evidence that demonstrates the influence of peripheral nerve injury on NMDA receptor-mediated synaptic transmission in the spinal dorsal horn. Using whole cell tight-seal methods, NMDA receptor-mediated excitatory postsynaptic currents (NMDA EPSCs) were recorded from superficial dorsal horn neurons in adult mouse spinal cord slices. Peripheral nerve injury-induced changes in the pharmacological and electrophysiological properties of synaptic NMDA receptors were studied. The ratio of the amplitude of NMDA EPSCs to that of non-NMDA EPSCs was larger in nerve-ligated neuropathic mice than in sham-operated control mice. The decay phase of the NMDA EPSCs was slower in nerve-ligated neuropathic mice. The NR2B subunit-specific NMDA receptor antagonist ifenprodil (10 microM) reduced the amplitude of the NMDA EPSCs and shortened their decay phase. The sensitivity of NMDA EPSCs to ifenprodil was significantly larger in nerve-ligated neuropathic mice than in sham-operated control mice. Single-cell RT-PCR analysis performed on superficial dorsal horn neurons showed that the incidence of NR2A mRNA-expressing neurons was reduced in nerve-ligated neuropathic mice. This result, together with the electrophysiological findings, suggests that the subunit composition of the subsynaptic NMDA receptors in the superficial dorsal horn was altered by peripheral nerve injury. Pharmacological and electrophysiological changes observed in the present experiments might be the underlying causes of the hyperalgesia and allodynia induced by peripheral nerve injury and inflammation.

D-amino-acid oxidase is involved in D-serine-induced nephrotoxicity.

D-serine is nephrotoxic in rats. Based on circumstantial evidence, it has been suspected that D-amino-acid oxidase is involved in this nephrotoxicity. Since we found that LEA/SENDAI rats lacked D-amino-acid oxidase, we examined whether this enzyme was associated with D-serine-induced nephrotoxicity using the LEA/SENDAI rats and control F344 rats. When d-propargylglycine, which is known to have a nephrotoxic effect through its metabolism by D-amino-acid oxidase, was injected intraperitoneally into the F344 rats, it caused glucosuria and polyuria. However, injection of d-propargylglycine into LEA/SENDAI rats did not cause any glucosuria or polyuria, indicating that D-amino-acid oxidase is definitely not functional in these rats. D-serine was then injected into the F344 and LEA/SENDAI rats. It caused glucosuria and polyuria in the F344 rats but not in the LEA/SENDAI rats. These results indicate clearly that D-amino-acid oxidase is responsible for the D-serine-induced nephrotoxicity.

Differential expression of NMDA receptor subunits between neurons containing and not containing enkephalin in the mouse embryo spinal cord.

We transfected cultures of mouse spinal cord slices with the enhanced green fluorescent protein (GFP) gene driven by the promoter for preproenkephalin, using the particle-mediated gene transfer system adapted for small neurons in the superficial dorsal horn, and observations were made after 4-6 days in vitro. A considerable number of cells in the superficial dorsal horn were observed to express GFP fluorescence, reminiscent of the previously reported distribution of enkephalinergic neurons in the spinal cord. The number of GFP-expressing neurons increased in response to forskolin application. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of single neurons revealed that the N-methyl-d-aspartate (NMDA) receptor NR2B subunit is expressed more frequently in enkephalinergic neurons, and the NR2A subunit more frequently in non-enkephalinergic neurons. These observations suggest that expression of NMDA receptor subunits is controlled differentially in distinct populations of neurochemically identified neurons in the spinal cord. Biolistic particle-mediated gene transfection seems useful for identifying neuronal phenotypes in organotypic cultures of the spinal cord.

Spatial learning and long-term potentiation of mutant mice lacking D-amino-acid oxidase.

We evaluated the role of D-amino-acid oxidase on spatial learning and long-term potentiation (LTP) in the hippocampus, since this enzyme metabolizes D-amino-acids, some of which enhance the N-methyl-D-aspartate receptor functions. The Morris water maze learning and the LTP in the CA1 area of the hippocampal slice were observed in wild-type mice and mutant mice lacking D-amino-acid oxidase. The mutant mice showed significantly shorter platform search times in the water maze and significantly larger hippocampal LTPs than the wild-type mice. These results suggest that the abundant D-amino-acids in the mutant mouse brain facilitate hippocampal LTP and spatial learning.

Facilitatory action of halothane at subanesthetic concentrations on glutamatergic excitatory synaptic transmission in the CA1 area of adult rat hippocampus.

Whole-cell recordings were made from pyramidal cells visually identified in the CA1 field of adult rat hippocampal slices, and the effects of subanesthetic concentrations of halothane on excitatory postsynaptic currents mediated by non-N-methyl-D-aspartate subtype glutamate receptors were investigated. Halothane concentrations were measured by gas chromatography. At concentrations of 0.2 mM and 0.6 mM, halothane reversibly decreased the amplitude of excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation of Schaffer collateral fibers, and the decrease was accompanied by enhanced paired-pulse facilitation, consistent with the previously reported presynaptic site of halothane's inhibitory action. By contrast, at lower concentrations (0.02 mM and 0.05 mM), halothane increased the amplitude of EPSCs without any appreciable changes in paired-pulse facilitation. Moreover, the frequency of miniature EPSCs arising spontaneously in the presence of tetrodotoxin (mEPSCs) was increased by subanesthetic halothane, but the amplitude of the mEPSCs did not change significantly. These observations suggest that at subanesthetic concentrations halothane postsynaptically enhances glutamatergic excitatory synaptic transmission. This may provide a vital clue to elucidation of the neural mechanisms of the nociceptive reflex enhancement and excitatory state that occur at light levels of anesthesia.

Gabapentin affects glutamatergic excitatory neurotransmission in the rat dorsal horn.

We investigated the effects of gabapentin (GBP) on glutamatergic synaptic transmission in the dorsal horn of the rat spinal cord. Patch clamp whole cell recordings were made from superficial and deep dorsal horn neurons of rat spinal cord slices. In the majority of neurons in the superficial lamina, GBP decreased the amplitudes of evoked excitatory postsynaptic currents (evoked EPSCs) mediated by either non-NMDA or NMDA receptors. In contrast, neurons in the deep lamina showed variable effects, with a lower incidence of decrease in amplitude of evoked EPSCs and a subset of neurons showing an increase in amplitude of evoked NMDA receptor-mediated EPSCs. No enhancement of evoked non-NMDA receptor-mediated EPSCs was observed in either lamina. To determine whether the observed effects of GBP are presynaptic and/or postsynaptic, spontaneous miniature excitatory postsynaptic currents (mEPSCs) were studied. In neurons that showed a decrease in its frequency of mEPSCs by GBP, no change in the amplitude or shape accompanied the effect. On the other hand, in neurons that showed an increase in the frequency of NMDA receptor-mediated mEPSCs, the effect accompanied an increase in amplitude. These results suggest that GBP presynaptically inhibits glutamatergic synaptic transmission predominantly in the superficial lamina, while postsynaptically enhancing NMDA receptor-mediated transmission in some neurons of the deep lamina. The antinociceptive effects of GBP may involve the inhibition of the release of excitatory amino acids from presynaptic terminals.