Evidence for NMDA receptor in the afferent synaptic transmission of the vestibular system

This study aimed to define the pharmacology and physiological role of the N-methyl-d-aspartate (NMDA) receptor in the synapse between the hair cells and primary afferent neurons in the vestibular system. The spontaneous and mechanically evoked spike discharges of vestibular nerve fibers were extracellularly recorded in isolated inner ear from the axolotl (Ambystoma tigrinum). Pressure ejection of NMDA (10⁻⁶ to 10⁻³ M) elicited a dose-dependent increase of the basal spike discharge from the vestibular nerve fibers. Extracellular magnesium antagonized the NMDA effect in a dose-dependent manner.d(-)-2-amino-5-phosphonovaleric acid (AP5, 10⁻⁵ to 10⁻³ M) and 7-chloro-kynurenic acid (7ClKyn, 10⁻⁶ to 10⁻³ M) inhibited the basal activity of the vestibular nerve fibers. 7ClKyn also diminished the responses elicited by the mechanical stimulation of the preparation. Glycine (10⁻⁹ to 10⁻⁶ M) applied by bath substitution enhanced the NMDA responses, and the glycine agonistd-serine partially reversed the 7ClKyn inhibitory action. These results suggest that NMDA receptors participate in the generation of the basal spike discharge of vestibular system primary afferent neurons, but its activation is not critical for the response to brief mechanical stimuli.     

Course and targets of the calbindin D-28k subpopulation of primary vestibular afferents

The subpopulation of primary vestibular afferents (PVA) displaying immunoreactivity for the calcium binding protein Calbindin D-28k (Calb) is constituted of particularly large bipolar neurons in the vestibular ganglion (VG) that innervate the central regions of the vestibular end organs exclusively via calyx endings on type I vestibular hair cells. These large-diameter PVA are characterized by irregular spontaneous discharge patterns and predominantly phasic firing properties with respect to natural vestibular stimulation. The present study describes the complete course and terminations of Calb+ PVA in the cerebellar cortex, the cerebellar (CN) and vestibular nuclei (VN) of the mouse. To eliminate the two sources of Calb+ fibers in the cerebellum, i.e., the Calb+ primary vestibular input and the axons of cerebellar Purkinje cells (PC), in their totality, a unilateral eighth nerve transection was performed in the PC-deficient mutant mice, Purkinje cell degeneration (pcd/pcd) and Lurcher (Lc/+). Neurectomy in these mutants results in a complete ipsilateral loss of Calb+ fibers in the cerebellar cortex, the CN and VN. The Calb+ primary vestibular input on the contralateral side terminates solely in the rostral half of the ventral uvula and in the nodulus of the cerebellar cortex. Calb+ fibers traverse all three subdivisions of the CN, but terminations were found only in the lateral and medial cerebellar nuclei. In the VN, Calb+ PVA terminations were restricted to the superior, the ventral part of the lateral, the lateral portion of the medial, and the inferior vestibular nuclei. Calb+ terminals were also present in the small cell group Y and Cajal's interstitial nucleus of the vestibular nerve as well as in defined areas of the reticular formation. All Calb+ PVA are strictly unilateral. The results show that the Calb+ subpopulation of VG neurons is the sole source of Calb+ fibers and terminals in the PC-deficient cerebellum and the VN. The central input of this distinct subgroup of PVA is distributed in narrow posterior vermal areas and parts of the CN and VN. The cerebellar mutants, Purkinje cell degeneration and Lurcher, provide excellent tools to selectively investigate the subgroup of Calb+ PVA in the mouse in its entirety. J. Comp. Neurol. 402:111–128, 1998. © 1998 Wiley-Liss, Inc.     

Dye coupling does not explain functional crosstalk within dorsal root ganglia

Most primary sensory neurons in rat dorsal root ganglia (DRGs) are depolarized during repetitive impulse activity in neighboring neurons that share the same ganglion. We wondered whether this functional crosstalk might be mediated by a network of cytoplasmic bridges (gap junctions) between neighboring neurons and their satellite glia. Neurobiotin was injected intracellularly in whole excised DRGs. Some of the animals were intact, and others underwent transection of the ipsilateral sciatic nerve 7 to 21 days prior to injection. A total of 44 directly injected neurons were recovered histologically. There was little or no evidence of dye spread to neighboring satellite cells or neurons that would have indicated the presence of cytoplasmic bridges, certainly not enough to account for the nearly universal functional coupling that occurs among these neurons. Functional crosstalk within DRGs must therefore employ a different mechanism.     

Gap junctions couple astrocytes but not neurons in dissociated cultures of rat suprachiasmatic nucleus

Individual neurons dissociated from rat suprachiasmatic nucleus can express independently phased circadian firing rhythms in culture. The phases of these rhythms are unperturbed by reversible blockade of neuronal firing lasting 2.5 days, indicating that multiple circadian clocks continue to operate in the absence of conventional synaptic transmission. The possibility remains, however, that these circadian rhythms might depend on some other form of intercellular communication. In the present study, a potential role for gap junctional coupling in SCN cultures was evaluated by introduction of the tracer molecule Neurobiotin into both neurons (n = 98) and astrocytes (n = 10), as well as by immunolabeling for specific connexins, the molecular components of gap junctions. Astrocytes were extensively coupled to each other by connexin43-positive gap junctions, but no evidence was found for coupling of neurons to each other or to astrocytes. These data support the hypothesis that neurons expressing independently phased circadian rhythms in SCN cultures (‘clock cells’) are autonomous, single cell circadian oscillators, but do not exclude a role for glia in synchronizing neuronal clock cells in vivo.     

Neuropeptides and γ-Aminobutyric acid in the vestibular nuclei of the rat: an immunohistochemical analysis. I. Distribution

The distributions of substance P, Leu-enkephalin and γ-aminobutyric acid (GABA) containing structures in the rat vestibular nuclei were investigated by means of an indirect immunofluorescent method using specific antisera to substance P, Leu-enkephalin and glutamic acid decar☐ylase (GAD), respectively.Numerous positive neurons and fibers containing these three substances were found in the medical vestibular nucleus. Most of them were situated in the caudal part of the nucleus and those in the rostral part were concentrated dorsally. In the descending vestibular nucleus, a large number of substance P, Leu-enkephalin and GAD containing neurons were evenly distributed among longitudinally directing fiber bundles. A number of positive fibers with these substances were also observed. The lateral vestibular nucleus contained numerous coarse GAD-immunoreactive fibers surrounding Deiters' neurons, while substance P-immunoreactive and Leu-enkephalin-immunoreactive fibers were rather poorly distributed in this nucleus as well as in the superior vestibular nucleus.     

Cervical primary afferent input to vestibulospinal neurons projecting to the cervical dorsal horn: An anterograde and retrograde tracing study in the cat

Vestibulospinal neurons in the caudal half of the medial and descending vestibular nuclei terminate in the cervical spinal cord, not only in the ventral horn and intermediate zone but also in the dorsal horn. The purpose of the present study was to examine whether the areas containing these vestibulospinal neurons are reached by cervical primary afferents. In one group of experiments, wheat germ agglutinin-horseradish peroxidase conjugate and horseradish peroxidase were pressure injected into spinal ganglia C₂-C₈ and revealed anterogradely labeled fibers and boutons in the caudal part (caudal to the dorsal cochlear nucleus) of the ipsilateral medial and descending vestibular nuclei. This projection was verified in experiments in which wheat germ agglutinin-horseradish peroxidase conjugate was microiontophoretically injected into the caudal half of either the medial or the descending vestibular nuclei and revealed retrogradely labeled cells only in ipsilateral spina ganglia C₂-C₇, with a maximum of cells in C₃. In another group of experiments, after microiontophoretic injections of Phaseolus vulgaris leucoagglutinin or Biocytin into either the medial or the descending vestibular nuclei, anterogradely labeled fibers and boutons were present in the cervical spinal cord, mainly bilaterally in the dorsal horn (laminae I–VI) but also, to a lesser extent, in the ventral horn and intermediate zone. The existence of a loop that relays cervical primary afferent information to vestibulospinal neurons projecting to the cervical spinal cord, in particular the dorsal horn, may have implications for vestibular control over local information processing in the cervical dorsal horn. © 1995 Wiley-Liss, Inc.     

Neuronal gap junctions and morphologically mixed synapses in the spinal cord of a teleost, Sternarchus albifrons (gymnotoidei)

Spinal cord motoneurons in the gymnotid, Sternarchus albifrons, were studied electron microscopically with special reference to the freeze-fracture method. Two types of motoneurons were identified. Electromotor neurons are monopolar and are located in a midline column dorsal to the ventral gray. These cells have a small fraction of their surface covered by synapses from descending axons, often at nodes. The synapses have multiple gap junctions, but few presynaptic vesicles or other correlates of chemical transmission. The gap junctions have an ordinary appearance in freeze-fracture replicas and exhibit a highly ordered substructure. The not infrequent appositions between the cell bodies of electromotor neurons exhibit no junctional specializations. Ordinary motoneurons are multipolar and densely covered with axosomatic and axodendritic synapses. In thin sections these synapses can be divided into two groups according to whether the vesicles are spherical or flattened. Gap junctions occur only at the first type, thus forming ‘morphologically mixed’ synapses. In freeze-etch replicas of motoneurons, the gap junctions are often found near clusters of postsynaptic E face particles elsewhere associated with excitatory chemical transmission. In addition, vesicle attachment sites occur in the presynaptic membranes of some synapses with gap junctions. The morphological observations are consistent with dual chemical and electrical transmission at these particular synapses, i.e. electrical excitation across gap junctions and chemical excitation at active zones with spherical vesicles and post-synaptic E face particles.     

Primary afferent fibers establish dye-coupled connections in the frog central nervous system

Neurobiotin and Lucifer yellow, indicators of gap junctional coupling, were applied to primary afferent fibers of the frog. Following application of tracers to cervical or lumbar dorsal root fibers, a large number of labeled granule cells were detected in the corpus cerebelli, the brainstem, and the spinal cord. The vestibular nerve was found to be in dye-coupled connection with the granule cells of the auricular lobe of the cerebellum. After application of the tracers to the trigeminal nerve, elicited dye-coupled neurons located mainly in the termination area of the descending limb of the mesencephalic trigeminal nucleus. In control experiments with biotinylated dextrane amine, only primary afferent fibers were labeled. Our results suggest that gap junctional coupling exists between primary afferent fibers and their postsynaptic targets in the frog.     

Small neurons in the vestibular nerve root project to the marginal shell of the anteroventral cochlear nucleus in the cat

We injected biotinylated dextran amine (BDA) into marginal shell regions of the anteroventral cochlear nucleus (AVCN) of the cat. These injections led to retrograde labeling of cells including small cells (median soma area = 111 μm², equivalent diameter = 11.9 μm) in the vestibular nerve root (VNR), just ventral to an anterior part of the AVCN. This is an unexpected new finding. The cells were scattered among BDA-labeled fibers and were oriented parallel to the course of the VNR fibers. We suggest that the small neurons of the VNR might serve as second-order vestibular neurons conveying information from vestibular end organs to the cochlear nucleus (CN) and/or act as interneurons between the olivocochlear fibers in the VNR and the CN.     

Distribution of primary vestibular fibers in the brainstem and cerebellum of the monkey

Attempts were made to determine the central projections of ganglion cells innervating individual semicircular ducts in the monkey by implanting or injecting tritiated amino acids (leucine and/or proline), or horseradish peroxidase (HRP), selectively into a single ampulla. Central transport via the vestibular ganglion in animals receiving isotope implants or injections fell into three categories: (1) transport from ganglion cells innervating all receptive elements of the labyrinth, (2) transport from ganglion cells innervating the three semicircular ducts, and (3) transport from cells of the inferior vestibular ganglion innervating the posterior semicircular duct. Transneuronal transport of isotope was observed in secondary vestibular fibers in animals where proline was used and survival exceeded 12 days. Transneuronal labeling of secondary auditory fibers was independent of the [³H]amino acid used, and occurred with survivals of 10 or more days. HRP implanted into the ampulla of the lateral semicircular duct in several animals produced retrograde transport to efferent vestibular and cochlear neurons, but did not result in transganglionic labeling of primary vestibular or auditory fibers.Primary vestibular fibers terminate throughout the superior (SVN) and medial vestibular nuclei (MVN). Within SVN, terminals are most pronounced in its central large-celled portion, but extend into peripheral parts of the nucleus, except for a small medial area near its junction with the oral pole of MVN. Primary projections to MVN are homogenously distributed throughout the nucleus excepting a small circular area of sparse terminals along its ventral margin. Primary vestibular afferents terminate mainly in rostral and caudal portions of the inferior vestibular nucleus (IVN), but do not reach cell group ‘f’. Projections to the lateral vestibular nucleus (LVN) are restricted to its ventral part. Primary projections to the accessory vestibular nuclei reach the interstitial nucleus of the vestibular nerve (NIVN) and cell group ‘y’. Fibers project beyond the vestibular nuclei (VN) to terminate ipsilaterally in the accessory cuneate nucleus (ACN), the subtrigeminal lateral reticular nucleus (SLRN), and well-defined portions of the reticular formation (RF). Projections to SVN and MVN are derived primarily from ganglion cells innervating the semicircular ducts, while projections to caudal IVN, cell group ‘y’ and ACN are related mainly to macular portions of the vestibular ganglion. NIVN receives both macular and duct afferents. Posterior duct afferents terminate in medial portions of SVN, in rostrolateral portions of MVN, and in rostral IVN.Transneuronal transport of isotope increases the volume of terminal label in the ipsilateral VN, but not in dorsal LVN, or cell groups ‘f’ or ‘x’. The quality of transneuronal transport in secondary vestibular fibers is dependent upon: (1) survival time, (2) proximity to the VN, and (3) the excitatory or inhibitory nature of the projection.Primary vestibulocerebellar fibers terminate heavily in the ipsilateral nodulus and ventral uvula. Lesser projections reach the flocculus, deep folia of vermal lobules V and VI, and the lingula. Primary vestibulocerebellar projections terminate as mossy fiber rosettes in the granular layer of these cortical areas. No primary vestibular fibers terminate in the primate fastigial nuclei.     

Transneuronal transport of intracellularly injected biotinamide in primary afferent axons

Transneuronal transport of biotinamide was observed following intracellular injection of biotinamide into rat jaw-muscle spindle afferent axons. Microelectrodes were advanced into the mesencephalic nucleus of the trigeminal nerve where jaw-muscle spindle afferent axons were identified by their increased firing during stretching of the jaw-elevator muscles. Biotinamide (Neurobiotin) was then injected into individual axons and the animals were maintained under anesthesia for 2−6 h. The animals were then killed via an overdose of anesthetic and the brainstem was processed histochemically. Biotinamide-filled axon collaterals and terminals were readily visible in the trigeminal motor nucleus, the trigeminal sensory nuclei, and adjacent reticular formation. In addition to these intracellularly stained axons, two to five neurons per animal (total of 36 in eight rats) were observed with a homogenous gray reaction product distributed throughout their somata, proximal, and secondary dendrites. These neurons ranged in size from small (8–20 μm, n = 26) to medium-sized (<30 μm, n = 10) and were closely apposed by numerous (up to 20) biotinamide-stained spindle afferent boutons. Most of these neurons (n = 22) were located in the dorsomedial portion of the spinal trigeminal subnucleus interpolaris (Vi) 2.5−4.5 mm caudal to the intra-axonal injection site. Electron microscopic analysis in two rats suggests that the transneuronal biotinamide labeling occurred predominately through asymmetric, axodendritic synapses between biotinamide-filled axon terminals and Vi neuronal dendrites. Although recent in vitro studies have reported that biotinamide permeates through gap junctions, in this study we found no evidence of biotinamide traversing the gap junctions which exist between trigeminal mesencephalic nucleus (Vme) neuronal somata. These results demonstrate that biotinamide can occasionally be transneuronally transported presumably via synapses; further information is needed to explain the seemingly sporadic nature of this transport.     

Glutamate as a primary afferent neurotransmitter in the medial vestibular nucleus as detected by in vivo microdialysis

An in vivo microdialysis study using α-chloralose-anesthetized cats was performed to elucidate whether glutamate is actually released from the vestibular nerve terminals in the medial vestibular nucleus (MVN) with electrical stimulation of the vestibular nerve. When repetitive stimuli composed of rectangular pulses (200 μs in duration, 0.5 mA, and 0.1–50 Hz) were applied to the vestibular nerve for 10 min, a significant frequency-dependent increase in the release of glutamate was observed in the MVN. However, the levels of other amino acids such as aspartate, glycine and GABA remained unaltered with the stimuli. These findings indicate that glutamate is the primary afferent neurotransmitter from the vestibular nerve to the MVN neurons.     

Interferon regulatory factor 1 is induced by interferon-γ equally in neurons and glial cells

Class I MHC protein is induced in glia but not mature neurons by IFN-γ. We have compared IFN-γ signal transduction in these populations. There were identical levels of STAT1 homodimers and IRF-1 by gel-shift and IRF-1 mRNA was induced equally. However class I MHC, β2-microglobulin and interleukin 1-β converting enzyme mRNA levels were greatly reduced in neurons. These experiments show that there is no defect in expression of IRF-1 in response to IFN-γ in mature mouse neurons but that insufficient class I MHC gene expression is induced for detectable cell surface protein expression.     

Astrocytic dye coupling in rat hippocampus: Topography,developmental onset,and modulation by protein kinase C

Previous studies have shown that astrocytes constitute a functional syncytium whereas the cytoplasmata of individual cells are connected via gap junctions. Many studies have used cultured astrocytes and have examined electrical coupling with the help of double electrode techniques. Another approach has been the immunohistochemical detection of gap junction proteins in sections of brain tissue. From the results of these experiments it is difficult to infer the extent of astrocyticcoupling in situ. To get an impression of the distribution of coupled astrocytes we took advantage of the hippocampal slice preparation which leaves the topography of neurons and astrocytes intact. We performed injections of low molecular weight dyes into single electrophysiologically identified astrocyes. As these dyes can pass through gap junctions this leads to the staining of all connected cells in a certain area, limited by the diffusional spread of the dye. The results show that there is virtually no border to astrocytic coupling between the diverse hippocampal subdivisions. This widespread coupling could already be detected at postnatal day 4, the earliest age tested. Activation of protein kinase C with phorbol esters showed that it is possible to reduce gap junctional communication by some extent. We conclude that although no compartmentalization was seen with respect to astrocytic coupling, the intercellular communication of these glial cells has the capability of being regulated for example by neuronal signals which activate the phospholipase C pathway in astrocytes. © 1994 Wiley-Liss, Inc.     

Distribution of neurotransmitters, neuropeptides, and receptors in the vestibular nuclei complex of the rat: An immunocytochemical, in situ hybridization and quantitative receptor autoradiographic study

This article investigates the distribution of neurotransmitters, neuropeptides, and related receptors in the vestibular nuclei complex (VNC) of the adult rat by means of immunohistochemistry, in situ hybridization, and quantitative receptor autoradiography. The entire complex proves to be rich in muscarinic receptors and it shows a high density of imipramine and benzodiazepine binding sites. Peptidergic neurons and a few positive fibers are described in the caudal part of the VNC. In particular, the medial vestibular nucleus contains a number of neurons expressing both the enkephalin mRNA and peptide. This nucleus and the lateral vestibular nucleus are also rich in opiate receptors. Substance P, thyrotropin releasing hormone, and neurotensin receptors are also found in the medial and in the spinal vestibular nuclei. In spite of the presence of α₂ catecholaminergic receptors, no thyrosine-hydroxylase-immunoreactive elements are seen in the caudal VNC. The possible functional meaning of these data is discussed.     

Neuronal organization of the vestibulospinal system in the cat

In the lateral vestibular nucleus, vestibulospinal tract (VST) neurons were surveyed with microelectrodes in cats anesthetized with sodium pentobarbital. The VST neurons (n = 450) were classified by their properties; axonal courses (LVST and MVST). spinal segmental levels of their axonal termination (C1-3, C4-8, T1-13, L1-4, and L5-neurons), their orthodromic activation by the primary vestibular nerve (second-order and non-second-order vestibular neurons), and their location in the LVN. Inhibitory and excitatory effects of cerebellar stimulation on these classified VST neurons were investigated. 84% (259/308) neurons were observed to receive cerebellar corticovestibular inhibition. The rate was high, and almost the same among classified neurons; C1-3 to L5-neurons, and second-order and non-second-order neurons. However, the rate with MVST neurons (69%) was significantly lower than with LVST cells (87%). These neurons which received cerebellar inhibition were distributed in all areas even deep in the rostroventral region of the LVN, while neurons which did not receive were distributed in the ventral region of the LVN. Electrical stimulation of ipsi- and contralateral fastigial nuclei evoked monosynaptic excitation of the classified VST neurons. Rate of occurrence of crossed fastigiovestibular excitation was higher with cervical neurons (86%) than with lumbar neurons (43%), and higher with second-order neurons (78%) than with non-second-order neurons (41%). Neurons which received monosynaptic excitation from crossed fastigiovestibular fibers were distributed in the ventral region of the LVN. In total, 73% of the neurons were identified to receive either ipsi- or contralateral fastigiovestibular excitation. The results indicated that there was relative scarcity of fastigiovestibular projections in the dorsal region of the LVN. Spinovestibular and other afferents to the LVN were also investigated.     

Ontogeny of electrophysiological properties and dendritic pattern in second-order chick vestibular neurons

The pattern of development of several subpopulations of second-order vestibular neurons was investigated by using intracellular recordings from chicken brain slices to define the timing of morphological and electrophysiological changes occurring at 3 critical ages. Two embryonic stages, embryonic day 13 (E13) and E15–16, and also newborn chicks were selected according to previous anatomical findings showing the differentiation of primary vestibular afferents and their synapses within a distinctive brainstem vestibular nucleus, the tangential nucleus. The responses of these cells to depolarizing and hyperpolarizing current pulses and their postsynaptic responses to vestibular nerve stimulation were recorded, while simultaneously biocytin was injected for subsequent morphogenetic analysis. From this study, developmental schedules of membrane properties, synaptic responses, and dendritic differentiation were established for the 2 cell populations of the tangential nucleus and other neurons located in the surrounding vestibular nuclei. Compared with all other second-order vestibular neurons, the principal cells of the tangential nucleus exhibited a distinctive schedule. Mainly, this includes their pattern of firing on depolarization, the shape and duration of the vestibular-evoked excitatory postsynaptic potential, and the time of onset of dendritic outgrowth. In regard to these observations, E15–16 appears to be a turning point in principal cell ontogeny, whereas these features occur earlier in development for other second-order vestibular neurons. These findings, which indicate that the principal cells may have distinct membrane properties at specific ages, are discussed in light of their unique vestibular innervation and the possible consequences for vestibular signal processing. J. Comp. Neurol. 384:621–633, 1997. © 1997 Wiley-Liss, Inc.     

Localization of substance P-like immunoreactivity in guinea pig vestibular endorgans and the vestibular ganglion.

The immunocytochemical distribution of substance P (SP) in guinea pig vestibular endorgans and the vestibular ganglion was investigated. Two kinds of SP-immunoreactive fibers were distinguished. Most were thick, and found around or beneath sensory hair cells. These SP-immunoreactive fibers were distributed predominantly on the slope of the crista and the peripheral region of the macula. By electron microscopy, we confirmed this type of SP-like immunoreactivity to be restricted within primary afferent neurons. Some vestibular ganglion cells also showed SP-like immunoreactivity, suggesting that SP is present in some primary afferent neurons, and is involved in afferent neurotransmission. The characteristic distribution of SP may indicate functional differences within each endorgan. The other group of immunoreactive nerve fibers, varicous thin fibers, could be found in the stroma of vestibular endorgans, nerve trunk, vestibular ganglion, and along blood vessels of the vestibular ganglion. These fibers may have a different origin, and have an influence on blood flow and certain other functions.     

Interactions between excitatory amino acids and tachykinins in the rat spinal dorsal horn

Whole-cell patch-clamp technique of freshly isolated rat spinal dorsal horn (DH) neurons, intracellular recording from DH neurons in a slice preparation, and high performance liquid chromatography with fluorimetric detection of release of endogenous glutamate and aspartate from spinal cord slice following activation of primary afferent fibers were employed to investigate interactions between excitatory amino acids (EAA) and tachykinins [substance P (SP) and neurokinin A (NKA)]. Potentiation of N-methyl-D-aspartate (NMDA)-, quisqualate (QA)- and α-amino 3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-, but not kainate-induced currents by SP and NKA was found. Spantide II, a claimed novel nonselective tachykinin antagonist, effectively blocked the SP (2 nM )-induced potentiation of the responses of DH neurons to NMDA. In the presence of glycine (0.1 μM), the SP-evoked increase of the NMDA-induced current was prevented. However, 7-chlorokynurenic acid (2 μM), a competitive antagonist at the glycine allosteric site of the NMDA receptor, led to the reestablishment of the SP effect. Brief high frequency electrical stimulation of primary afferent fibers produced a longlasting potentiation of presumed monosynaptic and polysynaptic excitatory postsynaptic potentials and sustained enhanced release of endogenous glutamate (218.3± 66.1 %) and aspartate (286.3 ± 58.0%). Possible functional implications of the observed phenomena are discussed in relation to transmission and integration of sensory information, including pain.