Excitatory Amino Acid Agonists Research Articles

Histamine (H 3) receptors modulate the excitatory amino acid receptor response of the vestibular afferents

Although the effectiveness of histamine-related drugs in the treatment of peripheral and central vestibular disorders may be explained by their action on the vestibular nuclei, it has also been shown that antivertigo effects can take place at the peripheral level. In this work, we examined the actions of H 3 histaminergic agonists and antagonists on the afferent neuron electrical discharge in the isolated inner ear of the axolotl. Our results indicate that H 3 antagonists such as thioperamide, clobenpropit, and betahistine (BH) decreased the electrical discharge of afferent neurons by interfering with the postsynaptic response to excitatory amino acid agonists. These results lend further support to the idea that the antivertigo action of histamine-related drugs may be caused, at least in part, by a decrease in the sensory input from the vestibular endorgans. The present data show that the inhibitory action of the afferent neurons discharge previously described for BH is not restricted to this molecule but is also shared by other H 3 antagonists.

Rhythmic Motor Activity Evoked by NMDA in the Spinal Zebrafish Larva

We have examined the localization and activity of the neural circuitry that generates swimming behavior in developing zebrafish that were spinalized to isolate the spinal cord from descending brain inputs. We found that addition of the excitatory amino acid agonist N-methyl-d-aspartate (NMDA) to spinalized zebrafish at 3 days in development induced repeating episodes of rhythmic tail beating activity reminiscent of slow swimming behavior. The neural correlate of this activity, monitored by extracellular recording comprised repeating episodes of rhythmic, rostrocaudally progressing peripheral nerve discharges that alternated between the two sides of the body. Motoneuron recordings revealed an activity pattern comprising a slow oscillatory and a fast synaptic component that was consistent with fictive swimming behavior. Pharmacological and voltage-clamp analysis implicated glycine and glutamate in generation of motoneuron activity. Contralateral alternation of motor activity was disrupted with strychnine, indicating a role for glycine in coordinating left-right alternation during NMDA-induced locomotion. At embryonic stages, while rhythmic synaptic activity patterns could still be evoked in motoneurons, they were typically lower in frequency. Kinematic recordings revealed that prior to 3 days in development, NMDA was unable to reliably generate rhythmic tail beating behavior. We conclude that NMDA induces episodes of rhythmic motor activity in spinalized developing zebrafish that can be monitored physiologically in paralyzed preparations. Therefore as for other vertebrates, the zebrafish central pattern generator is intrinsic to the spinal cord and can operate in isolation provided a tonic source of excitation is given.

Multiple neurotransmitter receptors contribute to the spinal Fos expression

The aim of this study is to identify the receptors which could potentially mediate the activation of c-Fos. Therefore, the effects of neurotransmitter receptor agonists in the activation of c-Fos in spinal neurons were studied by intrathecal injection of excitatory amino acid (EAA) receptor agonists: N-Methyl- d-Aspartate (NMDA), ( S)-α-Amino-3-Hydroxy-5-Methyl-4-Isoxazolepropionic acid (AMPA), 2-Carboxyl-3-carboxmethyl-4-isopropenylpyMidine (Kainic acid, KA), (1S-3R)-1-Aminocyclopentane-1, 3-dicarboxylacid (ACPD), and substance-P receptor (neurokinin-1) agonist, [Sar 9, Met (O 2) 11] SP (SarMet-SP). All drugs tested activated the production of c-Fos in spinal dorsal horn neurons. AMPA was found as the most potent agonist tested producing market production of c-Fos particularly in neurons of lamina II at doses of 10 pM per 10-μl injection. At this dose, other agonists were relatively ineffective. At higher doses, AMPA significantly increased the activated cells. NMDA significantly increased c-Fos production to a marked extent only at doses above 10 nM per 10-μl injection. KA and ACPD were least potent of the excitatory amino acid agonists. Injection of SarMet-SP at doses of 1 nM activated Fos selectively in neurons of lamina I. A dose-dependent increase in number of c-Fos-positive cells was observed for AMPA, KA, ACPD, and SarMet-SP, whereas NMDA gave a very strong expression after a high dose with no dose dependency. These finding suggest that multiple neurotransmitter receptors lead to c-Fos production in spinal neurons.

Effect of excitatory amino acid receptor agonists on penile erection after administration into the CA3 hippocampal region in the rat

Objectives To examine whether hippocampal injection of specific agonists for glutamate ionotropic (NMDA, AMPA) and metabotropic (ACPD) receptors modulates erectile response in adult male rats. The paraventricular nuclei of the hypothalamus and hippocampus are believed to play a role in the central regulation of erectile function. Methods Four groups of 5 male Sprague-Dawley rats each were anesthetized and placed in a stereotaxic apparatus after carotid artery catheterization for measurement of arterial pressure and insertion of a 25-gauge needle in the corpus cavernosum for measurement of intracavernous pressure (ICP). The rats underwent hippocampal injections of saline (1 μL), NMDA (10 nmol/L), AMPA (10 nmol/L), or ACPD (10 nmol/L) followed by perfusion with saline and 10% formalin solutions. Brain slices 24 μm thick were mounted and stained with cresyl violet to identify the drug injection sites. Results Hippocampal injection of glutamate receptor subtype agonists produced multiple episodes of ICP elevation. NMDA injection was associated with a greater maximal ICP increase, duration of action, and mean onset of action latency than AMPA injection. ACPD injection had no statistically significant impact on ICP or arterial pressure changes. Histologic evaluation confirmed the injected sites were the CA3 fields of the hippocampus. Conclusions Our results suggest hippocampal glutamate receptors may be involved in initiation of penile erection. Ionotropic receptors appear to play a significant role in these regulatory mechanisms. NMDA receptors appear to be more potent than AMPA receptors, which appear to mediate fast excitatory synaptic transmission.

Peripheral and Central Sensitization: Expert Commentary on Selected Abstracts

Central Trigeminal Sensitization May be Mediated by Nonmeningeal Nociceptive Trigeminal Afferents and Influenced by Sex HormonesWe have previously shown that injection of the inflammatory irritant and small‐fiber excitant mustard oil (MO) into the temporomandibular joint (TMJ) region can reflexively induce a prolonged increase in the activity of both digastric and masseter muscles in rats. It is possible that peripheral excitatory amino acid (EAA) receptors play a role in this effect, because MO‐evoked increases in jaw muscle activity are attenuated by preapplication of the noncompetitive NMDA receptor antagonist MK‐801 into the TMJ region. In the present study, the EAA receptor agonists glutamate, NMDA, kainate, and AMPA were applied locally to the TMJ region. Jaw muscle responses similar to those evoked by MO application to the TMJ region were achieved with glutamate, NMDA, AMPA, and kainate. Repeated application of glutamate, NMDA, or AMPA at intervals of 30 min evoked responses in the ipsilateral jaw muscles that were of comparable magnitude. Co‐application of the NMDA receptor antagonist DL‐2‐amino‐5‐phosphonovalerate (0.5 μmol) significantly reduced the magnitude of the glutamate‐ and NMDA‐evoked ipsilateral jaw muscle responses without affecting responses evoked by AMPA. In contrast, co‐application of the non‐NMDA receptor antagonist 6‐cyano‐7‐nitroquinoxaline‐2,3‐dione (1 nmol) significantly reduced the magnitude of the glutamate‐ and AMPA‐evoked ipsilateral jaw muscle responses without affecting responses evoked by NMDA. This evidence suggests that both NMDA and non‐NMDA EAA receptor types are located within the TMJ region and may contribute to jaw muscle activity that can be reflexively evoked from the TMJ region.

Aspartate and glutamate mimetic structures in biologically active compounds.

Glutamate and aspartate are frequently recognized as key structural elements for the biological activity of natural peptides and synthetic compounds. The acidic side-chain functionality of both the amino acids provides the basis for the ionic interaction and subsequent molecular recognition by specific receptor sites that results in the regulation of physiological or pathophysiological processes in the organism. In the development of new biologically active compounds that possess the ability to modulate these processes, compounds offering the same type of interactions are being designed. Thus, using a peptidomimetic design approach, glutamate and aspartate mimetics are incorporated into the structure of final biologically active compounds. This review covers different bioisosteric replacements of carboxylic acid alone, as well as mimetics of the whole amino acid structure. Amino acid analogs presented include those with different distances between anionic moieties, and analogs with additional functional groups that result in conformational restriction or alternative interaction sites. The article also provides an overview of different cyclic structures, including various cycloalkane, bicyclic and heterocyclic analogs, that lead to conformational restriction. Higher di- and tripeptide mimetics in which carboxylic acid functionality is incorporated into larger molecules are also reviewed. In addition to the mimetic structures presented, emphasis in this article is placed on their steric and electronic properties. These mimetics constitute a useful pool of fragments in the design of new biologically active compounds, particularly in the field of RGD mimetics and excitatory amino acid agonists and antagonists.

Excitatory amino acid receptor activation in the raphe pallidus area mediates prostaglandin-evoked thermogenesis

To investigate the role of excitatory amino acid neurotransmission within the rostral raphe pallidus area (RPa) in thermogenic and cardiovascular responses, changes in sympathetic nerve activity to brown adipose tissue (BAT), BAT temperature, expired CO 2, arterial pressure, and heart rate were recorded after microinjection of excitatory amino acid (EAA) receptor agonists into the RPa in urethan-chloralose-anesthetized, ventilated rats. To determine whether EAA neurotransmission within the RPa is necessary for the responses evoked by disinhibition of the RPa or by prostaglandin E 2 acting within the medial preoptic area, BAT sympathetic nerve activity, BAT temperature, expired CO 2, arterial pressure, and heart rate were measured during these treatments both before and after blockade of EAA receptors within the RPa. Microinjection of EAA receptor agonists into the RPa resulted in significant increases in all measured variables; these increases were attenuated by prior microinjection of the respective EAA receptor antagonists into the RPa. Microinjection of prostaglandin E 2 into the medial preoptic area or microinjection of bicuculline into the RPa resulted in respective significant increases in BAT sympathetic nerve activity (+approximately 190% and +approximately 235% of resting levels), in BAT temperature (approximately 1.8 °C and approximately 2 °C), in expired CO 2 (approximately 1.1% and approximately 1.1%), and in heart rate (approximately 97 beats per minute (bpm) and approximately 100 bpm). Blockade of ionotropic EAA receptors within the RPa by microinjection of kynurenate completely reversed the prostaglandin E 2 or bicuculline-evoked increases in all of the measured variables. Blockade of either N-methyl- d-aspartate (NMDA) receptors or non-NMDA receptors alone resulted in marked attenuations of the prostaglandin E 2-evoked effects on all of the measured variables. These data demonstrate that activation of an EAA input to the RPa is necessary for the BAT thermogenic and the cardiovascular effects resulting from the actions of prostaglandin E 2 within the medial preoptic area or from the disinhibition of local neurons in the RPa.

Effect of excitatory amino acid receptor agonists on penile erection after administration into paraventricular nucleus of hypothalamus in the rat

ObjectivesTo investigate whether the excitatory amino acid receptor agonists can activate the paraventricular nucleus of the hypothalamus (PVN) and induce penile erections in the rat. MethodsMale adult Sprague-Dawley rats were used. A 26-gauge needle was inserted into the corpus cavernosum to monitor the intracavernous pressure (ICP) simultaneously with the systemic arterial pressure and heart rate. The study was divided into seven parts: stereotaxic delivery of N-methyl-d-aspartic acid (NMDA) (50 ng/500 nL) into the PVN; administration of the NMDA noncompetitive antagonist MK-801 (100 ng/250 nL) and NMDA (50 ng/250 nL) into the PVN; administration of the NMDA competitive antagonist (±)-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP; 100 ng/250 nL) and NMDA (50 ng/250 nL) into the PVN; microinjection of (±)-α-(amino)-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA; 100 ng/500 nL) into the PVN; microinjection of trans-(±)-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD; 100 ng/500 nL) into the PVN; saline 500 nL into the PVN; and intracavernous administration of NMDA (50 ng/100 μL). ResultsOn administration of NMDA into the PVN, a significant increase occurred in the ICP from a resting 8.3 ± 1.8 mm Hg to a peak at 59.0 ± 8.4 mm Hg. No change occurred in the resting ICP after administration of either the mixture of MK-801 and NMDA or CPP and NMDA into the PVN. Microinjection of AMPA, ACPD, or saline into the PVN and intracavernous administration of NMDA were all ineffective to induce an increase in ICP. ConclusionsThe results of this study suggest that ionotropic excitatory amino acid (NMDA) may have an effect on inducing penile erection through activation of the PVN in the rat.

PH modulates the vestibular afferent discharge and its response to excitatory amino acids

In the isolated inner ear of the axolotl (Ambystoma tigrinum )a cid pH decreased and basic pH increased the resting and mechanically evoked spike discharge of semicircular canal aierent neurons. Variations in pH also modi¢ed the aierent neuron response to N-methyl-D-aspartic acid (NMDA) acid and to (7)-a-amino-3-hydroxy-5-methylisoxazole- 4 -propionic acid (AMPA). Responses to both excitatory amino acid agonists increased at pH 7.8 (41% and 22%, respectively) and decreased by perfusion of the preparation with a saline solution, of pH 7.0 (28% in both cases). These results indicate that vestibular endorgans have a signi¢cant sensitivity to pH that could play a signi¢cant role in various pathological states, and may also contribute to the post-transductional processing of sensory information. NeuroReport14:1327^1328 � c 2003 Lippincott Williams & Wilkins.

GABAergic systems modulate nicotinic receptor-mediated seizures in mice.

The pharmacology of nicotinic receptor-mediated seizures was investigated in C3H mice. Eleven nicotinic agonists and six antagonists were administered centrally (i.c.v.). Epibatidine and epiboxidine were the most potent agonists tested, whereas acetylcholine and the alpha7*-selective compounds 3-(2,4-dimethoxybenzylidene)-anabaseine (GTS-21) and anabasine, were the least potent. Nicotine-induced seizures were blocked by cotreatment with either the nonselective antagonist mecamylamine or the alpha7*-selective antagonist methyllycaconitine. The alpha4beta2*-selective antagonist dihydro-beta-erythroidine was ineffective at blocking seizures. However, high doses of all six antagonists tested were fully efficacious in producing seizures, with d-tubocurarine being the most potent and mecamylamine the least potent. Potential relationships between nicotinic receptor-mediated seizures and drug effects on GABA function were also investigated. No correlation was seen between potencies of the agonists in producing seizures and stimulating [3H]GABA release or between potencies of the antagonists in producing seizures and antagonist inhibition of nicotine-stimulated [3H]GABA release. However, a robust correlation was detected between potencies of the agonists in producing seizures and the IC50 values for inhibition of nicotine-stimulated [3H]GABA release produced by agonist-induced receptor desensitization. We also compared inbred mouse strain sensitivity to nicotine, picrotoxin, bicuculline, and kainate-induced seizures. Robust positive correlations were revealed for nicotine-induced seizures and seizures induced by either picrotoxin or bicuculline, both GABAA receptor antagonists. No correlation was found between nicotine-induced seizures and those induced by the excitatory amino acid receptor agonist kainate. Based on these findings, we present a model for nicotinic receptor-mediated seizures mediated through GABAergic systems.

Perturbations in three medullary nuclei enhance fractionated breathing in awake goats.

Our aim was to determine the frequency and characteristics of a fractionated pattern of diaphragm and upper airway muscle activity and airflow during wakefulness and sleep in adult goats. A fractionated breath (FBr) was defined as three or more brief (40-150 ms) interruptions in the diaphragm activity not associated with multiple swallows, eructation, mastication, or movement. During a FBr, the discharge pattern in the diaphragm and upper airway muscles showed complete cycles of inspiration and expiration. Whereas the interval between peak diaphragm activity of the breath preceding the FBr to the first diaphragm peak of the FBr was 15-20% less than the average interval of the preceding five control breaths, the breath-to-breath interval of the five breaths after a FBr did not differ from the control breaths before the FBr event. In normal goats, FBr was evident in only 4 of 18 (22%) awake goats and in only one of these goats during non-rapid eye movement sleep. In 35 goats with implanted microtubules in the medulla, FBr were present in 14 (40%) goats. In these goats with FBr, 78% (11 of 14) had one or more implantations into or near the facial, vestibular, or raphe nuclei. The effect of perturbations in these nuclei is probably nonspecific, because injections into these nuclei with mock cerebrospinal fluid or excitatory amino acid-receptor agonist or antagonist produced both increases and decreases in the frequency of the FBr while not altering their characteristics. Finally, a swallow occurred at the termination or during the first breath after 60% of the FBr. We speculate that the FBr manifest 1) the disruption of a neuronal network, which coordinates breathing and other functions (such as swallowing), utilizing the same anatomic structures, and/or 2) transient changes in synaptic inputs that increase the rate of the normal respiratory rhythm generator or allow an ectopic, anomalous generator to become dominant.

Opioid receptors mediate a postsynaptic facilitation and a presynaptic inhibition at the afferent synapse of axolotl vestibular hair cells

This study was designed to determine the effects of opiate drugs on the electrical activity of afferent neurons and on the ionic currents of hair cells from semicircular canals. Experiments were done on larval axolotls ( Ambystoma tigrinum). The multiunit spike activity of afferent neurons was recorded in the isolated inner ear under both resting conditions and mechanical stimulation. Ionic currents were recorded using voltage clamp of hair cells isolated from the semicircular canal. In the isolated inner-ear preparation, microperfusion of either non-specific opioid receptor antagonist naloxone (10 nM to 1 mM), μ receptor agonist [D-Ala 2, N-Me-Phe 4,Gly 5-ol]-enkephalin (1 pM to 10 μM), or κ receptor antagonist nor-binaltorphimine (10 nM to 100 μM) elicited a dose-dependent long-lasting (>5 min) increase of the electrical discharge of afferent neurons. The μ receptor agonist funaltrexamine (1 nM to 100 μM) and the κ receptor agonist U-50488 (1 nM to 10 μM) diminished the basal spike discharge of vestibular afferents. The δ receptor agonist D-Pen 2-D-Pen 5-enkephalin (1 nM to 10 mM) and the antagonist naltrindole (1 nM to 10 mM) were without a significant effect. The only drug that displayed a significant action on hair-cell ionic currents was trans-(±)-3,4-dichloro- N -methyl- N -(2-[1-pyrrolidinyl]-cyclohexyl) benzeneacetamide methanesulfonate (U-50488) that reduced the Ca 2+ current in a dose-dependent fashion. On its own, μ receptor agonist [D-Ala 2, N-Me-Phe 4,Gly 5-ol]-enkephalin (0.01 and 10 μM) significantly potentiated the response of afferent neurons to the excitatory amino acid agonist (±)-α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (0.1 μM), while synaptic transmission was blocked by the use of high-Mg 2+, low-Ca 2+ solutions. Our data indicate that the activity of vestibular afferent neurons may be regulated in a complex fashion by opioid receptors: μ opioid receptors mediating an excitatory, postsynaptic modulatory input to afferent neurons, and κ receptors mediating an inhibitory, presynaptic input to hair cells.

Role of Ghrelin in the Control of Growth Hormone Secretion in Prepubertal Rats: Interactions with Excitatory Amino Acids

Ghrelin is a 28-amino-acid peptide, with an essential n-octanoyl modification at Ser3, that elicits growth-hormone (GH) secretion in rats and humans. At present, the mechanisms of ghrelin action and its interactions with other systems controlling GH secretion remain poorly characterized. In this context, the present study was undertaken to obtain information about ontogeny and possible gender differences in the GH-releasing activity of ghrelin, and to delineate its primary site(s) of action at the hypothalamus and/or pituitary. In addition, the interactions between ghrelin and other relevant signals in the control of GH secretion, such as excitatory amino acids (EAAs), nitric oxide (NO) and serotonin, were assessed. Experiments were carried out in infantile-prepubertal animals, when GH pulsatility is not yet established. Systemic administration of ghrelin (25 nmol/rat, i.p.) to 5-, 10- and 23-day-old male and female rats increased plasma GH levels from day 10 onwards. This action was NO dependent, since it disappeared in 23-day-old males after pretreatment with an inhibitor of NO synthase (NAME). Similarly, central infusion of ghrelin (3 nmol/rat, i.c.v.) elicited GH responses in 10- and 23-day-old animals significantly higher than after systemic administration. By contrast, in vitro challenge of pituitary tissue with increasing doses of ghrelin (10^–9–10^–7M) failed to enhance GH release into the incubation medium, whereas stimulation with GH-releasing hormone (GHRH; 10^–7M) or GHRP-6 (10^–7M) was effective. Finally, effects of ghrelin were blocked by pretreatment with MK-801 and NBQX antagonists of EAA ionotropic receptors and after manipulation of endogenous serotoninergic tone. In addition, the potent releasing activity of EAA agonists NMDA and AMPA was blunted by pretreatment with D-Lys3-GHRP-6, a selective antagonist of the cognate ghrelin receptor, i.e. the GH-secretagogue receptor. In conclusion, our results demonstrate that GH-releasing activity of ghrelin appears early in the infantile period, is NO dependent and involves a primary hypothalamic site of action. The data also demonstrate for the first time the existence of a cross-talk between ghrelin and other neurotransmitter systems, such as EAAs and serotonin, in precise control of GH secretion.

Localization of premotoneurons for an NMDA-induced repetitive rhythmical activity to TMNs.

The localization or characteristics of the premotoneurons for trigeminal rhythmical activity have not been clarified. We investigated the localization of premotoneurons generating an NMDA-induced repetitive rhythmical activity to trigeminal motoneurons (TMNs). The minimal circuitry for this rhythmical activity was determined using a fragmented slice preparation of the isolated brain stem from neonatal rats (0-3 days old). We recorded rhythmical neural activities from TMNs using whole and fragmented brainstem slices preparation including the trigeminal motor nucleus in the presence of the excitatory amino acid agonist NMA and the GABAA receptor antagonist, bicuculline methiodide (BIC). TMNs receive projections from premotoneurons for an NMDA-induced rhythmical activity, which can be located in the area 300 microm surrounding the trigeminal motor nucleus. NMA (20 microM) and BIC (10 microM) induced repetitive rhythmical activities on TMNs.

Contributions from rostral medullary nuclei to coordination of swallowing and breathing in awake goats

The purpose of this study was to determine whether neurons in the facial (FN), gigantocellularis reticularis (RGN), and vestibular (VN) nuclei contribute to the regulation of breathing, swallowing, and the coordination of these two functions. Microtubules were chronically implanted bilaterally in goats. Two weeks later during wakefulness, 100-nl unilateral injections were made of mock cerebral spinal fluid or an excitatory amino acid receptor agonist or antagonists. When the agonist, N-methyl-D-aspartic acid, was injected into any nuclei, breathing and swallowing increased transiently (15-30%; P < 0.05), whereas only injections of the antagonist 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo-(f)quinoxaline into VN increased swallowing (20%; P < 0.05). The phase of breathing in which the swallows occurred was not altered by any injections. However, more importantly, injections of the agonist and the antagonists significantly altered (P < 0.05) by 5-50% the respiratory phase-dependent timing and tidal volume effect of swallows on breathing relative to mock cerebral spinal fluid injections. In addition, these effects were not uniform for all three nuclei. We conclude that the FN, RGN, and VN are part of a neural circuit in the rostral medulla that regulates and/or modulates breathing, swallowing, and their coordination in the awake state.

Effects of cholinergic and noradrenergic agents on locomotion in the mudpuppy (Necturus maculatus).

Some neurotransmitters act consistently on the central pattern generator (CPG) for locomotion in a wide range of vertebrates. In contrast, acetylcholine (ACh) and noradrenaline (NA) have various effects on locomotion in different preparations. The roles of ACh and NA have not been studied in amphibian walking, so we examined their effects in an isolated spinal cord preparation of the mudpuppy ( Necturus maculatus). This preparation contains a CPG that produces locomotor activity when N-methyl- D-aspartic acid (NMDA), an excitatory amino acid agonist, is added to the bath. The addition of carbachol, a long acting ACh agonist, to the bath disrupted the walking rhythm induced by NMDA, while not changing the level of activity in flexor and extensor motoneurons. Adding clonidine, an alpha(2)-noradrenergic agonist, had no effect on the NMDA-induced walking rhythm. Physostigmine, an ACh-esterase inhibitor, disrupted the walking rhythm, presumably by potentiating the effects of endogenously released ACh. Atropine, an ACh antagonist that binds to muscarinic ACh receptors, blocked the effects of carbachol, indicating that the action is mediated, at least in part, by muscarinic receptors. In the absence of carbachol, atropine had no effect. Locomotion was not induced by carbachol, atropine or clonidine in a resting spinal cord preparation. Cholinergic actions do not seem to be essential to the CPG for walking in the mudpuppy, but ACh may convert a rhythmic walking state to a more tonic state with occasional bursts of EMG activity for postural adjustments.

Interaction of excitatory amino acid agonists with cortical afterdischarges in developing rats.

To determine the role of excitatory amino acids (EAAs) in genesis of two types of epileptic afterdischarges. Cortical stimulation and recording electrodes were implanted in 12-, 18-, and 25-day-old rats. Epileptic afterdischarges were induced by rhythmic stimulation of sensorimotor cortex. The stimulation was repeated 6 times with 20-min intervals. Ten minutes after the first afterdischarge, N-methyl-d-aspartate, homocysteine, or kainic acid was injected. The doses were chosen individually for different age groups to be subconvulsive. Type and duration of afterdischarges as well as type and severity of motor correlates were evaluated. N-methyl-d-aspartate prolonged afterdischarges only in 12-day-old rats, whereas two other drugs did it in all age groups. Motor correlates of afterdischarges were changed to flexion seizures in 12-day-old rats after N-methyl-d-aspartate and homocysteine; in 25-day-old rats homocysteine led to generalized tonic-clonic seizures (i.e., both patterns seen after substantially higher doses of these drugs in nonstimulated rats). Seizures lasted tens of minutes. Kainic acid did not change the motor pattern in any age group, but nonconvulsive EEG seizures were recorded in the interstimulation periods mainly in 18- and 25-day-old rats. Increased transition into the limbic type of afterdischarges appeared only after homocysteine in 18- and 25-day-old rats. A mutual potentiation of epileptic phenomena was induced by two agents. The actions of N-methyl-d-aspartate and kainic acid differ in all age groups; the effects of homocysteine were identical with those of N-methyl-d-aspartate in 12-day-old rats but not later. Only homocysteine augmented transition into the limbic type of afterdischarges.

Alterations in responses of ventral pallidal neurons to excitatory amino acids after long-term dopamine depletion.

The present study explored the possibility that excitatory amino acid (EAA) sensitivity within the ventral pallidum (VP) is altered by long-term removal of dopamine (DA). Electrophysiological experiments were conducted in chloral hydrate-anesthetized rats 21 to 28 days after they received unilateral substantia nigra injections of the dopaminergic toxin 6-hydroxydopamine (6-OHDA). VP neurons increased firing at low microiontophoretic ejection currents of the EAA agonists N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA); however, high currents decreased action potential amplitude and rapidly caused cessation of neuronal firing. These responses likely reflected the induction of depolarization block for they were reversed by coiontophoresis of the hyperpolarizing transmitter gamma-aminobutyric acid (GABA) at ejection current levels that normally suppressed firing. The ability of NMDA and AMPA to induce such inactivation was greater in the VP of 6-OHDA-lesioned hemispheres, but unchanged in reserpinized rats, verifying that the alterations in responding to NMDA were the result of chronic, rather than acute, DA removal. The adaptations do not appear to be the consequence of a diminished GABAergic tone for the ability of bicuculline to increase firing (due to blocking a tonic GABAergic input) was not changed. However, low ejection currents of GABA that were insufficient to alter firing rate greatly attenuated the ability of NMDA to induce an apparent depolarization inactivation when coiontophoresed with NMDA onto VP neurons of the lesioned, but not the unlesioned, hemisphere. These studies show that chronic DA removal altered the EAA-induced amplitude-decreasing (i.e., the apparent depolarization inactivation) effects in VP neurons in the absence of a decrease in GABAergic tone.

Plasticity in excitatory amino acid receptor-mediated descending pain modulation after inflammation.

The role for excitatory amino acids (EAAs) in the rostral ventromedial medulla (RVM) in descending pain modulation after persistent noxious input is unclear. In an animal model of inflammatory hyperalgesia, we examined the effects of intra-RVM microinjection of EAA receptor agonists and antagonists on paw withdrawal and tail-flick responses in lightly anesthetized rats. N-Methyl-D-aspartate (NMDA) produced effects that depended upon the postinflammatory time period. At 3 h postinflammation, NMDA induced facilitation at a lower dose (10 pmol) and inhibition at a higher dose (1000 pmol). At 24 h postinflammation, NMDA (0.1-1000 pmol) produced a dose-dependent inhibition. The facilitation and inhibition, respectively, were attenuated significantly by the preadministration of an NMDA receptor antagonist, DL-2-amino-5-phosphonovaleric acid (APV) (10 pmol, P < 0.05), to the same site. Intra-RVM microinjection of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) (0.1-100 pmol) produced dose-dependent inhibition at both 3 and 24 h postinflammation that was blocked by the preadministration of an AMPA/kainate receptor antagonist, 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)quinoxaline (100 pmol, P < 0.05). Unexpectedly, AMPA-produced inhibition was also significantly attenuated by preadministration of APV (10 pmol, P < 0.05). Compared with 3 h postinflammation, both NMDA and AMPA showed a leftward shift in their dose-response curves at 24 h postinflammation. These results demonstrate that NMDA and AMPA receptors in the RVM are involved in the descending modulation after inflammatory hyperalgesia. There is a time-dependent increase in EAA neurotransmission in the RVM after inflammation and NMDA receptors play an important role in AMPA-produced inhibition.

Models of white matter injury: Comparison of infectious, hypoxic‐ischemic, and excitotoxic insults

White matter damage (WMD) in preterm neonates is strongly associated with adverse outcome. The etiology of white matter injury is not known but clinical data suggest that ischemia-reperfusion and/or infection-inflammation are important factors. Furthermore, antenatal infection seems to be an important risk factor for brain injury in term infants. In order to explore the pathophysiological mechanisms of WMD and to better understand how infectious agents may affect the vulnerability of the immature brain to injury, numerous novel animal models have been developed over the past decade. WMD can be induced by antenatal or postnatal administration of microbes (E. coli or Gardnerella vaginalis), virus (border disease virus) or bacterial products (lipopolysaccharide, LPS). Alternatively, various hypoperfusion paradigms or administration of excitatory amino acid receptor agonists (excitotoxicity models) can be used. Irrespective of which insult is utilized, the maturational age of the CNS and choice of species seem critical. Generally, lesions with similarity to human WMD, with respect to distribution and morphological characteristics, are easier to induce in gyrencephalic species (rabbits, dogs, cats and sheep) than in rodents. Recently, however, models have been developed in rats (PND 1-7), using either bilateral carotid occlusion or combined hypoxia-ischemia, that produce predominantly white matter lesions. LPS is the infectious agent most often used to produce WMD in immature dogs, cats, or fetal sheep. The mechanism whereby LPS induces brain injury is not completely understood but involves activation of toll-like receptor 4 on immune cells with initiation of a generalized inflammatory response resulting in systemic hypoglycemia, perturbation of coagulation, cerebral hypoperfusion, and activation of inflammatory cells in the CNS. LPS and umbilical cord occlusion both produce WMD with quite similar distribution in 65% gestational sheep. The morphological appearance is different, however, with a more pronounced infiltration of inflammatory cells into the brain and focal microglia/macrophage ("inflammatory WMD") in response to LPS compared to hypoperfusion evoking a more diffuse microglial response usually devoid of cellular infiltrates ("ischemic WMD"). Furthermore, low doses of LPS that by themselves have no adverse effects in 7-day-old rats (maturation corresponding to the near term human fetus), dramatically increase brain injury to a subsequent hypoxic-ischemic challenge, implicating that bacterial products can sensitize the immature CNS. Contrary to this finding, other bacterial agents like lipoteichoic acid were recently shown to induce tolerance of the immature brain suggesting that the innate immune system may respond differently to various ligands, which needs to be further explored.