Prolonged GABAB receptor-mediated synaptic inhibition in the cat spinal cord: an in vivo study

In pentobarbitone-anaesthetised spinal cats, a comparison was made of the effects of intravenous bicuculline hydrochloride, a GABA_A-receptor antagonist, and several (-)-baclofen (GABA_B-receptor) antagonists (CGP 35348, 46381, 56999A) on the prolonged inhibition of extensor-muscle monosynaptic reflexes, recorded from lumbar ventral roots, by brief or continuous tetanic stimulation of low-threshold afferent fibres of hindlimb flexor muscles. Two components of brief tetanus inhibition were detected. Whilst possibly of similar central latency, the inhibition associated with GABA_B receptors had a longer time course than that reduced by bicuculline. Furthermore, whereas bicuculline reduced primary afferent depolarization, generated by the inhibitory volleys, and detected as dorsal-root potentials, such potentials were generally enhanced by intravenous baclofen antagonists. The inhibition of reflexes during and after continuous (333 Hz) tetanic flexor-nerve stimulation appeared to be predominantly associated with the activation of GABA_B receptors. In the period following continuous tetanic flexor-nerve stimulation, during which monosynaptic extensor reflexes were reduced in amplitude, the action potentials of the intraspinal terminations of extensor-muscle group-Ia afferent fibres were reduced in duration, as detected by the time course of the recovery of the threshold to extracellular microstimulation following the arrival of an orthodromic impulse. A reduction in termination action-potential duration also accompanied the reduction by microelectrophoretic (-)-baclofen of the release of excitatory transmitter from group-Ia terminations, both presynaptic effects being blocked by microelectrophoretic baclofen antagonists. However, the reduction of the duration of the action potential of individual group-Ia terminations, which followed continuous flexor-nerve stimulation, was not sensitive to the baclofen antagonist CGP 55845A, but was diminished by bicuculline methochloride. Intravenously administered bicuculline hydrochloride, however, had little or no effect on the inhibition of reflexes following continuous flexor-nerve stimulation. These observations are discussed in the context of possible intraspinal pathways and pre- and postsynaptic mechanisms for GABA_A and GABA_B receptor-mediated inhibition of the monosynaptic excitation of spinal motoneurones and of the functional significance of central GABA_B receptor-associated inhibitory processes, given the relatively minimal effects on motor activity and behaviour produced by baclofen antagonists that penetrate the mammalian blood-brain barrier. Received: 2 July 1997 / Accepted: 30 January 1998     

Ptychodiscus brevis toxin-induced depression of spinal reflexes involves 5-HT via 5-HT3 receptors modulated by NMDA receptor

The involvement of 5-hydroxytryptaminergic (5-HT) system for the Ptychodiscus brevis toxin (PbTx)-induced depression of spinal reflexes was evaluated. The reflex potentials were recorded at ventral root by stimulating the corresponding dorsal root in neonatal rat spinal cord in vitro. Superfusion of PbTx (2.8-84microM) depressed the monosynaptic (MSR) and polysynaptic (PSR) reflexes in a concentration-dependent manner. The depression of the reflexes was maximal with 84microM of the toxin. Ondansetron (0.1microM), a 5-HT(3) receptor antagonist, blocked the PbTx-induced depression of MSR and PSR. Spiperone (a 5-HT(2A) antagonist) or ketanserin (5-HT(2A/2C) antagonist and also at 5-HT(1B/1D)) failed to block the PbTx-induced depression of the reflexes. The 5-HT concentration of the cords was increased by four-fold after exposure to PbTx (28microM) and the increase was not seen in the cords pretreated with dl-2 amino-5-phosphonovaleric acid (APV, a NMDA receptor antagonist). Superfusion of 5-HT or phenylbiguanide (PBG, a 5-HT(3) receptor agonist) also produced depression of the spinal reflexes in a concentration-dependent manner. The 5-HT-induced depression of reflexes was blocked by ondansetron but not by spiperone. The results demonstrate that the PbTx-induced depression of spinal reflexes involves 5-hydroxytryptamine via 5-HT(3) receptors modulated by NMDA receptor.     

Antidromic conditioning of reciprocally inhibited monosynaptic extensor and flexor reflexes in decrebrate cats

Summary The effect of previous antidromic conditioning of reciprocally inhibited monosynaptic flexor and extensor reflexes was found to depend on the ventral root chosen. Use of a part of the root in which the test reflex was found led to increased inhibition, use of a neighboring root to disinhibition. It is suggested that Renshaw cells in the agonist pool inhibit only those Ia inhibitory interneurons which affect the antagonist. A model for the interconnection of Renshaw cells, Ia inhibitory interneurons and the antagonistic motoneuron pools is proposed which fits not only the results presented here, but also those on single units. The results indicate the presence of occlusion of inhibitory effects and support the assumption that these effects are distributed uniformly within a motoneuron pool.These investigations were begun at the Max-Planck-Institut für experimentelle Medizin (Göttingen) and were supported by the Deutsche Forschungsgemeinschaft (SFB 33 Nervensystem und Biologische Information).     

The effects of postsynaptic inhibition on the monosynaptic reflex of the cat at different levels of motoneuron pool activity

Summary The motoneurons to the Soleus muscle in the decerebrate cat were activated by the crossed extensor reflex, elicited by stimulation of the contralateral common peroneal (CP) nerve. Monosynaptic reflexes were obtained from the Soleus motoneuron pool by stimulation of the cut L7-S1 dorsal roots. The amplitude of the reflex increased approximately linearly with the recruitment level of the motoneuron pool. Tonic postsynaptic inhibition was induced in the Soleus moto-neuron pool by repetitive antidromic stimulation of the Lateral Gastrocnemius (LG) and Medial Gastrocnemius (MG) nerves at a rate of 17–47 stimuli/s. This reduced the size of the monosynaptic reflex at rest by at least 40%. However, when the motoneurons were active, the amplitude of the monosynaptic reflex obtained during repetitive stimulation of the LG-MG nerve increased with the recruitment level along the same curve as the control reflexes. Thus, tonic postsynaptic inhibition of the motoneurons per se cannot control the amplitude of the monosynaptic reflex independently of the recruitment level of the motoneuron pool. These experimental results verify predictions from computer simulations and suggest by exclusion that presynaptic inhibition is needed to control the amplitude of the monosynaptic reflex independently of the recruitment level of the motor pool.     

Adenosine modulates excitatory synaptic transmission and suppresses neuronal death induced by ischaemia in rat spinal motoneurones

Although adenosine is an important neuromodulator, its role in modulating motor functions at the level of the spinal cord is poorly understood. In the present study, we investigated the effects of adenosine on excitatory synaptic transmission and neuronal death induced by experimental ischaemia by using whole-cell patch-clamp recordings from lamina IX neurones in spinal cord slices. Adenosine significantly decreased the frequency of miniature excitatory postsynaptic currents (mEPSCs) in almost all neurones examined that could be mimicked by an A₁ receptor agonist, N ⁶-cyclopentyladenosine (CPA), and inhibited by an A₁ receptor antagonist, 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX). Interestingly, adenosine increased mEPSC frequency in the presence of DPCPX in a subpopulation of neurones. In these neurones, an A_2A receptor agonist, 2-[4-(2-carbonylethyl)-phenethylamino]-5′-N-ethylcarboxamidoadenosine (CGS21680), increased mEPSC frequency. Adenosine also induced an outward current that was blocked by the addition of Cs⁺ and tetraethylammonium into the patch-pipette solution and inhibited in the presence of Ba²⁺. The adenosine-induced outward current was mimicked by CPA, but not CGS21680, and inhibited by DPCPX. Moreover, superfusing with ischaemia simulating medium (ISM) generated an agonal inward current in all of the neurones tested. The latencies of the inward currents induced by ISM were significantly prolonged by adenosine or CPA, but not by CGS21680. These results suggest that adenosine receptors are functionally expressed in both the pre- and postsynaptic sites of lamina IX neurones and that their activation may exert multiple effects on motor function. Moreover, this study has provided a cellular basis for an involvement of A₁ receptors in the neuroprotective actions of adenosine.     

Phosphinic acid derivatives as baclofen agonists and antagonists in the mammalian spinal cord: an in vivo study

The actions of a series of derivatives of 3-aminopropyl-phosphinic acid as baclofen agonists and antagonists have been examined on the synaptic excitation of neurones by impulses in primary afferent fibres in the lumbar spinal cords of pentobarbitone-anaesthetised cats and rats. Both the pre-and postsynaptic inhibitory actions of microelectrophoretic (-)-baclofen were reduced by similarly administered CGP 35 348, 36 742, 46 381, 52 432, 54 626 and 55 845, the latter being the most potent antagonist. None of these antagonists either decreased or increased the excitability of spinal neurones, and the inhibitory action of GABA was reduced only by local concentrations of antagonists which also reduced the action of piperidine-4-sulphonic acid, a GABA_A agonist. Although the weak inhibitory effect of 3-aminopropylphosphinic acid in both the rat and the cat was not reduced by these baclofen antagonists, the pre-and postsynaptic inhibitory effects of 3-aminopropyl-methyl-osphinic acid (CGP 35 024), which was more potent than (-)-baclofen, were reduced by the antagonists. Like (-)-baclofen, CGP 35 024 was relatively ineffective in reducing transmitter release in the cord from the terminals of excitatory spinal interneurones, the terminals of excitatory tracts in the dorsolateral funiculus and the cholinergic terminals of motor axon collaterals. In both rat and cat cords, receptors for (-)-baclofen could not be demonstrated to be activated by microelectrophoretic GABA, possibly because of the predominantly dendritic location of GABA_B receptors. Spinal pre-and postsynaptic baclofen receptors appeared to be pharmacologically similar but differed from those in the higher central nervous system of the rat, where 3-aminopropylphosphinic acid has been reported to be an effective baclofen agonist. The compounds tested, particularly CGP 55 845 and 46 381, will be of use in further investigations of the physiological relevance of baclofen receptors at central synapses where GABA may be the transmitter.     

缺氧对谷氨酸能和GABA能突触传递的影响

Neurons in the mammalian central nervous sysytem (CNS) are highly sensitive to the availability of oxygen. Hypoxia alters synaptic transmission in a few minutes. Both glutamatergic and γ-aminobutyric acid (GABA)ergic synaptic transmissions respond to hypoxic exposure with prominent modification. Glutamate receptors, GABA receptors, adenosine receptor, and some endogenous neuromodulators are involved in the preservation of neuron function. Since the neuroprotection in all hypoxic tolerant species examined so far relies on significant increase in GABA and decrease in glutamate, it may be an important strategy to make a moderate balance of glutamate/GABA synaptic transmission against hypoxic insults.     

Facilitation of synaptic activity in the frog spinal cord produced by 4-aminopyridine

Reflex activity in the isolated frog spinal cord is increased after adding 4-aminopyridine (4-AP) to the bath. Depression of synaptic transmission by increased magnesium or EGTA is effectively antagonized by 4-AP. It is suggested that 4-AP facilitates synaptic transmission in the central nervous system by interacting with the calcium mechanism involved in transmitter release from presynaptic nerve terminals.     

A new metabotropic glutamate receptor agonist: Developmental change of its sensitivity to receptors in the newborn rat spinal cord

A novel metabotropic glutamate receptor agonist, (2S,1′R,2′R,3′R)-2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV), reduced the monosynaptic excitation in newborn rat spinal cord rather than polysynaptic discharges at the nanomolar range without causing postsynaptic depolarization of motoneurones. Its inhibitory action on the monosynaptic excitation reduced in due course of time after birth. On the contrary, the inhibitory action of a metabotropic GABA_B receptor agonist, baclofen, did not show marked developmental change. DCG-IV should be expected to have the potential to provide further useful information on the physiological function of metabotropic glutamate receptors.     

Coeruleospinal inhibition of visceral nociceptive processing in the rat spinal cord

Visceral nociceptive information is transmitted in two different areas of the spinal cord gray matter, the dorsal horn and the area near the central canal. The present study was designed to examine whether visceral nociceptive transmission in the two different areas is under the control of the centrifugal pathways from the locus coeruleus/subcoeruleus (LC/SC). Extracellular recordings were made from the L(6)-S(2) segmental level using a carbon filament glass microelectrode (4-6 MOmega). Colorectal distentions (80 mmHg) were produced by inflating a balloon inside the descending colon and rectum. In both dorsal horn and deep area neurons, responses to colorectal distention were inhibited during electrical stimulation (30, 50 and 70 microA, 100 Hz, 0.1 ms pulses) of the LC/SC. It is well known that spinothalamic tract (STT) neurons excited by visceral nociceptive stimuli are located in the dorsal horn and that postsynaptic dorsal column (PSDC) neurons which conduct visceral nociceptive signals in the dorsal column (DC) are located near the central canal of the spinal cord. The present study, therefore, suggests that the descending LC/SC system can inhibit visceral nociceptive signals ascending through the STT and the DC pathways.     

Selective effects of (−)-baclofen on spinal synaptic transmission in the cat

Summary When ejected microelectrophoretically near spinal interneurones of cats anaesthetised with pentobarbitone and under conditions where postsynaptic excitability was maintained artificially at a constant level, (–), but not (+), -baclofen selectively reduced monosynaptic excitation by impulses in low threshold muscle (Ia and Ib) and cutaneous (A) afferents. Polysynaptic excitation of interneurones and Renshaw cells by impulses in higher threshold afferents was less affected, and baclofen had little or no effect on the cholinergic monosynaptic excitation of Renshaw cells. Glycinergic and gabergic inhibitions of spinal neurones were relatively insensitive to baclofen. These stereospecific actions of baclofen, produced by either a reduction in the release of excitatory transmitter or postsynaptic antagonism, suggest that Ia, Ib, and A afferents may release the same excitatory transmitter which differs from that of spinal excitatory interneurones.Microelectrophoretic (–), but not (+), -baclofen also reduced primary afferent depolarization of ventral horn Ia extensor afferent terminations produced by impulses in low threshold flexor afferents, without altering either the electrical excitability of the terminations or their depolarization by electrophoretic GABA or L-glutamate. This stereospecific action of baclofen is interpreted as a reduction in the release of GABA at depolarizing axo-axonic synapses on Ia terminals.     

Sensitivity of monosynaptic test reflexes to facilitation and inhibition as a function of the test reflex size: a study in man and the cat

Summary In parallel experiments on humans and in the cat it was investigated how the sensitivity of monosynaptic test reflexes to facilitation and inhibition varies as a function of the size of the control test reflex itself. In man the monosynaptic reflex (the Hoffmann reflex) was evoked in either the soleus muscle (by stimulation of the tibial nerve) or the quadriceps muscle (by stimulation of the femoral nerve). In the decerebrate cat monosynaptic reflexes were recorded from the nerves to soleus and medial gastrocnemius muscles; they were evoked by stimulation of the proximal ends of the sectioned L7 and S1 dorsal roots. Various excitatory and inhibitory spinal reflex pathways were used for conditioning the test reflexes (e.g. monosynaptic Ia excitation, disynaptic reciprocal inhibition, cutaneous inhibition, recurrent inhibition, presynaptic inhibition of the Ia fibres mediating the test reflex). It was shown that the additional number of motoneurones recruited in a monosynaptic test reflex by a constant excitatory conditioning stimulus was very much dependent on the size of the test reflex itself. This dependency had the same characteristic pattern whatever the conditioning stimulus. With increasing size of the test reflex the number of additionally recruited motoneurones first increased, then reached a peak (or plateau) and finally decreased. A similar relation was also seen with inhibitory conditioning stimuli. The basic physiological factors responsible for these findings are discussed. Finally, the implications for the interpretation of experiments in man with the H-reflex technique are considered.     

Extracellular potassium accumulation and transmission in frog spinal cord

In the isolated frog spinal cord, intracellular recordings from motoneurons and neuroglia and extracellular recordings from dorsal and ventral roots were used to compare electrophysiological changes produced by tetanic stimulation of dorsal roots with those resulting from increasing extracellular potassium concentration, [K⁺]₀. Many of the after-effects of tetanic dorsal root stimulation could be mimicked by increasing [K⁺]₀. These include the following: depolarization of motoneurons and neuroglia, prolongation and depression of excitatory postsynaptic potentials, depression of dorsal root potentials, facilitation and depression of ventral root potentials, as well as increases in spontaneous synaptic activity and depression of antidromic spikes recorded from motoneurons. The levels of [K⁺]₀ necessary to produce effects comparable to those following maximal dorsal root stimulation are about twice those estimated from measurements with K-selective microelectrodes or glial depolarization, suggesting that both these methods underestimate the amount of potassium which accumulates in the narrow intercellular spaces between neurons and glia in the intermediate region of the spinal cord.It is concluded that, at least within the isolated frog spinal cord, K⁺ accumulation is a significant factor modifying transmission following dorsal root tetanic stimulation and that its distribution is inhomogeneous.     

Formation of synaptic vesicles in the superior cervical ganglion of cat: choline dependency

Summary De novo formation of synaptic vesicles was studied electron microscopically in synapses of the superior cervical ganglion of cat. Following prolonged electrical stimulation in a choline deficient condition, almost complete loss of vesicle content was observed. After choline administration, together with the partial restoration of transmission, the reappearance of vesicles could be seen in nerve terminals. In interpreting our observations, the double role of choline (transmitter precursor and membrane constituent) is emphasized.     

缺氧使非洲电鱼小脑浦肯野细胞之间的GABA能突触传递长时程增强

目的:研究急性缺氧对非洲电鱼小脑浦肯野细胞(Pc)之间γ-氨基丁酸(GABA)能突触传递的影响。方法:采用配对全细胞膜片钳记录法,记录电鱼小脑Pc-Pc之间的抑制性突触后电流(IPSC),观察急性缺氧对Pc-Pc IPSC的影响,以及GABA_A受体拮抗剂和谷氨酸α-氨基-3-羟基-5-甲基-4-异噁唑丙酸(AMPA)受体拮抗剂对Pc-Pc IPSC缺氧反应的调节作用。结果:短暂缺氧使Pc-Pc IPSC的幅值显著增大,表现为长时程增强(LTP);GABA_A受体拮抗剂荷包牡丹碱逆转了Pc-Pc IPSC的LTP,表现为长时程抑制;AMPA受体拮抗剂6-氰基-7-硝基喹喔啉-2,3-二酮(CNQX)阻断了Pc-Pc IPSC的LTP,表现为短时程增强。结论:急性缺氧引起电鱼小脑Pc之间的GABA能突触活动持续增强,GABAA受体和AMPA受体共同介导这种反应,提示GABA能和谷氨酸能突触活动的平衡可能是电鱼以及其他缺氧耐受动物缺氧保护反应的关键机制。     

Mechanisms mediating the brain stem control of somatosensory transmission in the dorsal horn of the cat's spinal cord: an intracellular analysis

Summary The effect of brainstem stimulation was studied on neurones recorded intracellularly in the superficial and deeper laminae of the lumbosacral dorsal horn of the spinal cord in anaesthetised cats. Stimulation in the nucleus locus coeruleus (LC) produced a hyperpolarisation in 4/13 multireceptive neurones and produced a biphasic action consisting of a hyperpolarisation which was followed by a depolarisation in 3/13 neurones. These actions were produced irrespective of whether the multireceptive neurone was located in the superficial or deeper laminae of the dorsal horn. Stimulation failed to produce postsynaptic potentials in the remaining 6/13 multireceptive neurones. The amplitude of hyperpolarisation was increased by the passage of depolarising pulses through the recording microelectrode and decreased by hyperpolarising pulses. Stimulation in other brainstem areas such as, the lateral (FTL), paralemniscal (FTP) and central (FTC) divisions of the tegmental field and the nuclei raphe magnus (NRM) and reticularis magnocellularis (RMc) also hyperpolarised neurones in the dorsal horn. The polarity of hyperpolarisation evoked from some brainstem areas (FTP, FTC, RMc) could be reversed to depolarisation by the passive diffusion of ions from the recording microelectrode containing 3M-KCl. Brainstem (LC, NRM, FTP, FTL) stimulation generated long lasting (700 ms) hyperpolarisation on 4/4 selectively nocireceptive neurones of lamina I. There was, however, no effect on the activity of 5/5 neurones recorded in laminae I/II which in addition to receiving excitatory cutaneous inputs were inhibited by heat stimuli. Stimulation in LC also produced dorsal root potentials (DRPs) and reduced the amplitude of simultaneously recorded excitatory postsynaptic potentials (EPSPs) generated by the activation of primary afferent fibres in 3 multireceptive neurones. It is concluded that inhibition of nociceptive transmission in the spinal cord from LC and other brainstem areas may involve both pre- and postsynaptic mechanisms.     

Coerulospinal influence on recurrent inhibition of spinal motonuclei innervating antagonistic hindleg muscles of the cat

The locus coeruleus's (LC's) effect on recurrent inhibition of gastrocnemius-soleus (GS) and common peroneal (CP) monosynaptic reflexes (MSRs) was demonstrated to exceed the concomitant facilitation, indicating the independency of LC's disinhibition and facilitation measures in this study. In contrast, the disinhibition effect correlated closely with the recurrently inhibited MSRs. The disinhibition phenomenon was also accompanied by progressive delay and diminution in the Renshaw cell field potential. Hence, the recovery of recurrently inhibited MSRs was probably due, in part at least, to the LC's inhibition of the related Renshaw cell activity. Furthermore, the site-specific, discordant changes in the disinhibition of GS, compared with CP MSRs, as revealed by tracking studies imply that representations of these antagonistic motonuclei may occupy different LC loci. Accordingly, the nonuniform disinhibition may be due to the activation of discrete aggregates of LC neurons which are responsible predominantly in controlling the recurrent inhibitory pathway belonging to one or the other of the antagonistic motonuclei. These findings support a differential LC inhibitory control of Renshaw cell activity, releasing the related motoneurons for the Ia synaptic transmission — a disinhibitory process that is crucial for the LC's independent control of the recurrent circuit of antagonistics extensor and flexor motoneurons.     

Effects of physostigmine on the inhibition of trigeminal motoneurones by cutaneous impulses in the cat

Summary Physostigmine (less than 0.3 mg/kg, i. v.) was administered systemically to cats, and the effect studied on the electrical activities of trigeminal motoneurones. Physostigmine depressed the monosynaptic reflex evoked by stimulation of the mesencephalic trigeminal tract and recorded from the masseter nerve, and potentiated the inhibition of this monosynaptic reflex by volleys in skin nerves. This effect began within 1–2 min after the injection of physostigmine, reached a maximum at 5–12 min and thereafter decayed gradually. Skin nerve stimulation induced inhibitory postsynaptic potentials (IPSPs) in trigeminal motoneurones. After physostigmine administration these IPSPs were increased in amplitude, and were accompanied by a prolonged, late hyperpolarization. The early phase of this IPSP and the late hyperpolarization showed different sensitivities to the intracellular injection of chloride ions and to polarizing currents, indicating different mechanisms for their production. The effect of physostigmine was blocked by atropine (1 mg/kg, i.v.) but not by dihydro--erythroidine (0.5 mg/kg, i.v.).     

Effects of nociceptin and analogues of nociceptin upon spontaneous dorsal root activity recorded from an in vitro preparation of rat spinal cord

The 17 amino acid peptide nociceptin has been implicated in pain modulation in the central nervous system. The effects of bath applied nociceptin, and some analogues of nociceptin, upon spontaneous lumbar dorsal root activity have been investigated in an isolated preparation of rat spinal cord. Nociceptin was found to reversibly depress spontaneous dorsal root activity at concentrations of 1.0 μM and 10.0 μM (IC₅₀ 2.0 μM), whereas acetyl-nociceptin at concentrations up to 10 μM had no detectable effect. Omission of the last four amino acids (nociceptin 1–13), increased the potency of the effect upon dorsal root activity by approximately 100-fold (IC₅₀ 30 nM), but activity was lost when only the first seven amino acids of the nociceptin molecule (nociceptin 1–7) were tested.     

Involvement of group I metabotropic glutamate receptors and glutamate transporters in the slow excitatory synaptic transmission in the spinal cord dorsal horn

Our experiments demonstrate a novel role for group I metabotropic glutamate receptor (mGluR) subtypes 1 and 5 in generating a long-lasting synaptic excitation in the substantia gelatinosa (SG) and deep dorsal horn (DH) neurons of the rat spinal cord. In the present study we have investigated a slow excitatory postsynaptic current (EPSC), elicited by a brief high intensity (at Adelta/C fiber strength) and high frequency (20 or 100 Hz) stimulation of primary afferent fibers (PAFs) using whole-cell patch-clamp recordings from neurons located in the DH (laminae II-V) in spinal cord slices of young rats and wild-type and gene-targeted mice lacking mGluR1 subtype. The results shown here suggest that the activation of both mGluR1 and mGluR5 along with NK1 receptors, may be involved in the generation of the slow EPSC in the spinal cord DH. Inhibition of glial and neuronal glutamate transporters by dl-threo-beta-benzyloxyaspartate (TBOA) enhanced the group I mGluR-dependent slow EPSC about eightfold. Therefore, we conclude, that glutamate transporters strongly influence the group I mGluR activation by PAFs possibly at sensory synapses in the DH. Overall these data indicate that stimulus trains can generate a sustained and widespread glutamate signal that can further elicit prolonged EPSCs predominantly mediated by the group I mGluRs. These slow excitatory synaptic currents may have important functional implications for DH cell firing and synaptic plasticity of sensory transmission, including nociception.