Biomedical subjects

A J Lawrence

Publications and source records attributed to A J Lawrence.

At least 55 records · Page 3Linked to original sources

PubMed1999

Up-regulation of cortical AMPA receptor binding in the fawn-hooded rat following ethanol withdrawal.

The present study has employed quantitative receptor autoradiography to compare the binding of (S)-[3H]5-fluorowillardiine to (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors in the brains of alcohol-preferring Fawn-Hooded (FH) rats, alcohol non-preferring Wistar-Kyoto (WKY) rats, and FH rats following a 28-day period of 5% ethanol consumption with or without ethanol withdrawal. Significantly higher binding of [3H]5-fluorowillardiine was found in the cingulate cortex (+12%) and claustrum (+13%) in alcohol naïve FH rats compared to WKY rats. Chronic ethanol consumption decreased binding of (S)-[3H]5-fluorowillardiine in four cortical regions (frontal, parietal, occipital and temporal cortex), hippocampus and septohippocampal nucleus. In contrast, ethanol withdrawal induced a significant "rebound" increase in binding by +22% in frontal and parietal cortex, by +17% in cingulate cortex and +13% in claustrum, and by +14% in the septohippocampal nucleus compared to chronic ethanol-exposed FH rats. The findings suggest that AMPA receptors in frontal cortical regions are sensitive to ethanol and therefore may be implicated in the predisposition of alcohol preference in FH rats.

Alanine↗

PubMed1999

Visualisation of AMPA binding sites in the brain stem of normotensive and hypertensive rats.

The present study has employed in vitro receptor autoradiography with (S)-[(3)H]-5-fluorowillardiine (10 nM) to visualise the presence of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) binding sites in the brain stems of adult (16-18 weeks) normotensive (Wistar-Kyoto (WKY) and Don Ryu (DRY)) and Spontaneously Hypertensive (SHR) rats. Similar topographic distribution and density of (S)-[(3)H]-5-fluorowillardiine binding was observed in the nucleus tractus solitarius (NTS) of all three strains. Specific (S)-[(3)H]-5-fluorowillardiine binding sites were also visualised in sections of nodose ganglion from adult WKY rats, demonstrating that vagal afferent perikarya possess AMPA binding sites. However, while unilateral vagal deafferentation did not result in a significant decrease in binding site density in the caudal half of the rat NTS, the visualisation of AMPA binding sites on the nodose ganglion is consistent with the existence of a population of binding sites on vagal afferent terminals. In the caudal half of the rat NTS, AMPA binding sites appear to be predominantly postsynaptic in nature.

Alanine↗

PubMed1999

Expression of preprogalanin mRNA following acute and chronic restraint stress in brains of normotensive and hypertensive rats.

Exposure to stress is known to induce widespread changes in the central nervous system (CNS) involving multiple neuropeptides. The neuropeptide galanin has been implicated in the central response to different stressors; however, the role of galanin in the response to restraint stress has not been reported. Therefore, this study utilised in situ hybridisation histochemistry to observe the effects of acute and chronic restraint stress on preprogalanin (preproGAL) mRNA expression in the CNS of normotensive (Wistar Kyoto; WKY) and Spontaneously Hypertensive (SHR) rats. Rats were exposed to 1 h of restraint for 0 (control), 1, 3, 5, or 10 consecutive days, and central preproGAL mRNA expression following these restraint periods was compared between strains. Significant differences in the basal expression of preproGAL mRNA were detected, with expression decreased by approximately 50% in the supraoptic nucleus (SON; P<0. 01) and increased by approximately 100% in the rostral ventrolateral medulla (RVLM; P<0.05) of SHR when compared to WKY. Following acute restraint (1 session), preproGAL mRNA expression was significantly increased by approximately 135% in the central nucleus of the amygdala (CeA; P<0.05) in WKY. In SHR, significant increases of up to 300% were observed in the CeA (P<0.01) and SON (P<0.05) following chronic restraint (up to 10 days). In addition, expression of preproGAL mRNA was significantly decreased in the locus coeruleus (LC) of SHR following acute restraint (1 session) (P<0.05). These results provide the first evidence that both acute (LC) and chronic (CeA, SON) restraint stress is associated with alterations in preproGAL mRNA expression. As such, the present study provides further evidence linking neurons containing galanin with the central response to restraint stress.

Acute Disease↗

PubMed1999

The role of opioid-dopamine interactions in the induction and maintenance of ethanol consumption.

1. Alcohol is one of the most widely used recreational drugs, but also one of the most widely abused, causing vast economic, social and personal damage. 2. Several animal models are available to study the reinforcing mechanisms that are the basis of the abuse liability of ethanol. Innate differences in opioid or dopamine neurotransmission may enhance the abuse liability of ethanol, as indicated by animal and human studies. 3. Opioid antagonists have been shown to be effective, both experimentally and clinically, in decreasing ethanol consumption, presumably since ethanol induces the release of endogenous opioid peptides in vivo. However, ethanol may also stimulate the formation of opiate-like compounds, which could interact with opioid (or dopamine) receptors. Ethanol may cause changes in neurotransmission mediated via opioid receptors that determines whether alcohol abuse is more or less likely. 4. Ethanol appears to facilitate dopamine release by increasing opioidergic activity, disinhibiting dopaminergic neurons (by inhibition of GABAergic neurotransmission) via mu-opioid receptors in the ventral tegmental area (VTA) and delta-opioid receptors in the nucleus accumbens (NAcc). The effects of ethanol would be antagonised by presynaptic kappa-opioid receptors present on dopaminergic terminals in the NAcc. 5. Mesolimbic dopamine release induced by ethanol consumption seems to indicate ethanol-related stimuli are important, focussing attention on and enabling learning of the stimuli. However, studies indicate that there are redundant pathways, and neural pathways 'downstream' of the mesolimbic dopamine system, which also enable the reinforcing properties of ethanol to be mediated.

Alcohol Drinking↗

PubMed1999

Nitrergic stimulation of the locus coeruleus modulates blood pressure and heart rate in the anaesthetized rat.

To investigate whether nitric oxide is involved in the cardiovascular responses mediated via the locus coeruleus, the effects of microinjections of L-arginine and L-glutamate into the locus coeruleus on blood pressure and heart rate were investigated in sodium pentobarbitone-anaesthetized rats. Unilateral microinjection of L-arginine (25, 50 nmol) elicited dose-related depressor (-17 +/- 4, -25 +/- 4 mmHg) and bradycardic (13 +/- 3, 24 +/- 6 b.p.m.) effects. Furthermore, these effects were attenuated by prior local microinjection of N(G)-nitro-L-arginine (40 nmol). Peripheral muscarinic receptor blockade with atropine methyl nitrate (1 mg/kg, i.v.) attenuated the bradycardic but not the depressor responses to L-arginine. L-Glutamate (2 nmol) microinjections also mediated depressor (-27 +/- 6 mmHg) and bradycardic (53 +/- 23 b.p.m.) effects that were attenuated by microinjections of dizocilpine maleate (1 nmol) into the locus coeruleus. In addition, pretreatment with N(G)-nitro-L-arginine (40 nmol) also significantly attenuated the depressor response elicited by L-glutamate. These results suggest that nitrergic and glutamatergic pathways are operative within the locus coeruleus to modulate cardiovascular function, and also that a functional interaction may exist between the nitrergic and glutamatergic systems within the rat locus coeruleus.

Anesthesia, General↗

PubMed1999

Hypotension activates neuropeptide Y-containing neurons in the rat medulla oblongata.

The present study was designed to determine whether neurons within cardiovascular control nuclei of the rat brainstem that become activated following a hypotensive insult also possess the capacity to utilize neuropeptide Y. Adult male Wistar-Kyoto rats were injected with glyceryl trinitrate (10 mg/kg, i.p.) or vehicle, and 4 h later anaesthetized (pentobarbitone, 60 mg/kg, i.p.) and transcardially perfused. The brains were removed and processed by standard two-colour peroxidase immunohistochemistry. Activated cells were determined by incubation with a primary antibody to Fos protein, which was followed by a second incubation with a primary antibody to neuropeptide Y for double labelling of Fos-positive cells. Compared to vehicle, glyceryl trinitrate-induced hypotension caused a marked induction of Fos protein in the caudal one-third of the nucleus tractus solitarius (bregma -14 to -13.3 mm), which tailed off rapidly in more rostral sections. Following hypotension, significant populations of activated cells were also observed in the rostral and caudal ventrolateral medulla. In the caudal nucleus tractus solitarius and the posterior part of the medial nucleus tractus solitarius, respectively, 15 of 104 and 40 of 120 Fos-positive cells exhibited cytoplasmic neuropeptide Y immunoreactivity following hypotension, compared to seven of 40 and 15 of 40 in vehicle-treated rats, indicating a significant (two- to three-fold) increase in double-labelled cells following systemic glyceryl trinitrate (P < 0.05, unpaired t-test). In contrast, in the anterior part of the medial nucleus tractus solitarius, the number of double-labelled cells did not change following hypotension. An increase in double-labelled cells was also observed in the rostral ventrolateral medulla (2.5-fold increase compared to vehicle) and caudal ventrolateral medulla (5.8-fold increase compared to vehicle) following hypotension. These data indicate that, in the rat, neuropeptide Y-containing neurons are involved in the central response to a hypotensive challenge. The primary regions where neuropeptide Y-containing neurons appear to be activated are the caudal one-third of the nucleus tractus solitarius and the caudal ventrolateral medulla/rostral ventrolateral medulla, which are key nuclei associated with the integration of the baroreceptor heart rate reflex and sympathetic vasomotor outflow.

Animals↗

PubMed1999

Thymosin beta 4 sulfoxide is an anti-inflammatory agent generated by monocytes in the presence of glucocorticoids.

The possibility that glucocorticoids upregulate the expression of anti-inflammatory mediators is an exciting prospect for therapy in inflammatory diseases, because these molecules could give the therapeutic benefits of steroids without toxic side effects. Supernatants from monocytes and macrophages cultured in the presence of glucocorticoids increase the dispersion of neutrophils from a cell pellet in the capillary tube migration assay. This supernatant factor, unlike other neutrophil agonists, promotes dispersive locomotion of neutrophils at uniform concentration, lowers their adhesion to endothelial cells, inhibits their chemotactic response to fMLP and induces distinctive morphological changes. Here we show that thymosin beta4 sulfoxide is generated by monocytes in the presence of glucocorticoids and acts as a signal to inhibit an inflammatory response. In vitro, thymosin beta4 sulfoxide inhibited neutrophil chemotaxis, and in vivo, the oxidized peptide, but not the native form, was a potent inhibitor of carrageenin-induced edema in the mouse paw. Thymosin beta4 is unique, because oxidation attenuates its intracellular G-actin sequestering activity, but greatly enhances its extracellular signaling properties. This description of methionine oxidation conferring extracellular function on a cytosolic protein may have far-reaching implications for future strategies of anti-inflammatory therapy.

Amino Acid Sequence↗

PubMed1999

Activation of the acidic isoform of phospholipase A2 from Naja mossambica mossambica venom by oleoyl imidazolide requires the cooperative action of two ionizing groups.

The acidic phospholipase A2 isoform from the spitting cobra Naja mossambica mossambica is activated irreversibly by treatment with a molar equivalent of oleoyl imidazolide. The kinetics of the chemical modification of the enzyme can also be monitored by measuring the large reduction of tryptophan fluorescence, which is accompanied by a distinct red shift. The addition of a single molar equivalent of oleic acid to the enzyme produces an instantaneous reduction in fluorescence but with a barely detectable red shift, confirming that the response to oleoyl imidazolide results from covalent modification of the protein rather than hydrolysis of the reagent. The pH dependence of both activation and fluorescence reduction by oleoyl imidazolide has an optimum rate near pH 8.0. We propose that long-chain fatty acids and long-chain acyl imidazolides bind at a single activation site and that the reaction of the imidazolides involves two protein residues, one of which is a nonessential histidine residue and the other a primary amino group.

Animals↗

PubMed1999

Renal medullary interstitial infusion of norepinephrine in anesthetized rabbits: methodological considerations.

We tested methods for delivery of drugs to the renal medulla of anesthetized rabbits. Outer medullary infusion (OMI) of norepinephrine (300 ng. kg(-1). min(-1)), using acutely or chronically positioned catheters, reduced both cortical (CBF; 15%) and medullary perfusion (MBF; 23-31%). Inner medullary infusion (IMI) did not affect renal hemodynamics, whereas intravenous infusion reduced CBF (15%) without changes in MBF. During OMI of [(3)H]norepinephrine, much of the radiolabel (approximately 40% with chronically positioned catheters) spilled over systemically. Nevertheless, autoradiographic analysis showed the concentration of radiolabel was about fourfold greater in the infused medulla than the cortex. In contrast, during IMI, only approximately 5% of the infused radiolabel spilled over into the systemic circulation and approximately 64% was excreted by the infused kidney. The resultant intrarenal levels of radiolabel were considerably less with IMI compared with OMI. In rabbits, OMI therefore provides a useful method for targeting agents to the renal medulla, but given the considerable systemic spillover with OMI, its utility is probably limited to substances that are rapidly degraded in vivo.

Anesthesia↗

PubMed1999

Ethanol consumption by Fawn-Hooded rats following abstinence: effect of naltrexone and changes in mu-opioid receptor density.

BACKGROUND: Relapse after abstinence can be modelled in rats using an alcohol deprivation effect (ADE) of enhanced ethanol consumption after a period of enforced abstinence from ethanol; however, not all rat strains display such an effect. We wanted to examine the effect of naltrexone on ethanol consumption by ethanol-preferring Fawn-Hooded (FH) rats using such a model. METHODS: FH rats were given continual free-choice access to a 5% ethanol solution or water (4 weeks) followed by 2 weeks of water alone. At the end of this abstinence period, osmotic minipumps were implanted subcutaneously to deliver saline (n = 4) or naltrexone (n = 4; 8.4 mg/kg/day for 4 weeks). After recovery from surgery, the rats were again given access to 5% ethanol under the same free-choice conditions (4 weeks). A third group of age-matched controls drank only water during the behavioral trial. At the end of the behavioral trial, the rats were decapitated and an autoradiographic examination was made of micro-opioid receptor density through the forebrain using the ligand [125I]FK-33824. RESULTS: First, a period of enforced abstinence from ethanol consumption caused a significant (p < 0.05) and prolonged increase in ethanol preference (+18%) and decrease in water consumption (-53%), although the volume of ethanol consumed (ml/day) did not vary, indicating an atypical ADE in this rat strain. Second, naltrexone significantly (p < 0.05) decreased ethanol consumption by the FH rats in terms of absolute amount of ethanol consumed and preference for ethanol solution, but this effect of naltrexone diminished over time, concurrent with a robust and significant elevation in micro-opioid receptor density in all brain regions examined (p < 0.05). Finally, ethanol consumption alone also upregulated micro-opioid receptor density relative to nondrinking controls in a number of brain regions, which included the nucleus accumbens (+29%) and caudate-putamen (+15%,p < 0.05), but decreased micro-opioid receptor density in other regions including the substantia nigra pars reticulata, which was suggestive of an indirect effect on micro-opioid receptors. CONCLUSIONS: The data suggest that continual long-term naltrexone treatment may not be effective in the treatment of alcoholism, possibly because of the induced increase in micro-opioid receptor density.

Alcohol Drinking↗

PubMed1998

Targeted expression of a toxin gene to D1 dopamine receptor neurons by cre-mediated site-specific recombination.

Idiopathic Parkinson's disease involves the loss of midbrain dopaminergic neurons, resulting in the presynaptic breakdown of dopaminergic transmission in the striatum. Huntington's disease and some neurodegenerative diseases with Parkinsonian features have postsynaptic defects caused by striatal cell death. Mice were generated in which an attenuated form of the diphtheria toxin gene (tox-176) was expressed exclusively in D1 dopamine receptor (D1R)-positive cells with the aim of determining the effect of this mutation on development of the basal ganglia and on the locomotor phenotype. Transgenic mice expressing Cre, a site-specific DNA recombinase, were crossed with a second line in which a transcriptionally silenced tox-176 gene was inserted into the D1R gene locus by homologous recombination. Young doubly transgenic mutant mice expressing the tox-176 gene displayed bradykinesia, dystonia, and had falls caused by myoclonic jerks. The mutant brain had evidence of apoptosis and reactive gliosis and, consistent with the D1R expression pattern, the striatum was reduced in volume, and the Islands of Calleja were absent. In contrast, the cortex was of normal thickness. D1Rs were not detectable in mutants by in situ hybridization or ligand autoradiography, whereas D2 dopamine receptor (D2R) mRNA and protein was present in the striatum. In addition, substance P and dynorphin, neuropeptides known to be expressed in D1R-positive striatonigral projection neurons were not detectable. Enkephalin, a marker found in D2-positive striatopallidal projection neurons was expressed in the mutant brain. The mutant represents a novel neurodegenerative disease model with a dramatic extrapyramidal phenotype.

Animals↗

PubMed1998

Regional distribution of low affinity kainate receptors in brain of Macaca fascicularis determined by autoradiography using [3H](2S,4R)-4-methylglutamate.

Binding of [3H](2S,4R)-4-methylglutamate, a novel low affinity kainate receptor agonist, was studied in brain sections of a Macaca fascicularis monkey. In cerebellar sections, [3H](2S,4R)-4-methylglutamate bound to a single population of sites (KD = 20 nM) and was inhibited by various glutamate receptor ligands: kainate > 6-cyano-7-nitroquinoxaline-2,3-dione > L-glutamate >> AMPA. (S)-5-lodowillardiine and (RS)-2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl)propanoic acid (ATPA), drugs selective for the GluR5 subunit, displaced 50% and 40% of binding, respectively. Autoradiography revealed topographic binding of [3H](2S,4R)-4-methylglutamate. Binding in cortex was highest in layer 5 and restricted to CA2/3 in hippocampus. Levels of binding were high in septum and hypothalamus. Moderate densities of binding were found in caudate-putamen, cerebellar granular layer, nucleus tractus solitarius, cuneate nucleus and area postrema. Binding in spinal cord was concentrated in dorsal horn. [3H](2S,4R)-4-Methylglutamate shows differential binding throughout primate brain and is a valuable new ligand for low affinity kainate receptors.

Animals↗

PubMed1998

Nicotinic acetylcholine receptor mediated modulation of evoked excitatory amino acid release in the nucleus tractus solitarius of the rat: evidence from in vivo microdialysis.

In vivo microdialysis was used to measure release of endogenous l-glutamate and l-aspartate in the nucleus tractus solitarius of the anaesthetised rat evoked by baroreceptor loading. Aortic constriction, the method of loading, elicited a reproducible increase in extracellular levels of l-glutamate to 322+/-139% of basal levels, which could be attenuated by concomitant local administration of the nicotinic acetylcholine receptor antagonist mecamylamine (100 microM).

Animals↗

PubMed1998

Autoradiographic visualisation of axonal transport of adenosine A1 receptors along the rat vagus nerve and characterisation of adenosine A1 receptor binding in the dorsal vagal complex of hypertensive and normotensive rats.

The present study had employed in vitro receptor autoradiography with [3H]DPCPX to visualise the presence of adenosine A1 receptors on the rat nodose ganglion, which contains the perikarya of vagal afferent neurons projecting the the nucleus tractus solitarius (NTS). In addition, unilateral vagal ligation resulted in an accumulation of [3H]DPCPX binding adjacent to the ligatures, indication that adenosine A1 receptors are subject to axoplasmic flow along the rat vagus nerve. Radioligand binding assays were utilised to characterise the properties of adenosine A1 receptors in the dorsal vagal complex (NTS, area postrema and dorsal motor nucleus of the vagus) of pup and adult normotensive (Wistar Kyoto, WKY) and hypertensive (spontaneously hypertensive, SHR) rats. Saturation binding indicated that the affinity (KD) of [3H]DPCPX, and the binding site density (Bmax) were not different between the adult WKY and SHR, although the pup SHR had a lower KD value than the pup WKY rat. Competition binding assays revealed complex differences between the two rat strains; however, with respect to hypertension, the affinity of the selective adenosine A1 agonist, cyclohexyladenosine (CHA), was markedly reduced in the membranes from SHR (Ki approximately 93 nM) compared to WKY (approximately 6 nM). Such an observation is consistent with the attenuated responses of SHRs to intra-NTS injections of adenosine.

Adenosine↗

PubMed1998

Distribution of opioid peptide gene expression in the limbic system of Fawn-Hooded (alcohol-preferring) and Wistar-Kyoto (alcohol-non-preferring) rats.

Preprodynorphin and preproenkephalin mRNA expression was examined in the CNS of two rat strains, the alcohol-preferring Fawn-Hooded (FH) and the alcohol-non-preferring Wistar-Kyoto (WKY), using in situ hybridisation histochemistry. Relative to the WKY, the FH showed significantly lower levels of preproenkephalin mRNA in the striatum and nucleus accumbens (-24% and -17% respectively), but a higher level of preprodynorphin mRNA in the hippocampus (+33%). The depressed level of preproenkephalin mRNA in the nucleus accumbens may be implicated in alcohol-seeking behaviour.

Alcohol Drinking↗

PubMed1998

Bradykinin B2 receptors in nodose ganglia of rat and human.

The present study has employed in vitro electrophysiology to characterise the ability of bradykinin to depolarise the rat isolated nodose ganglion preparation, containing the perikarya of vagal afferent neurons. Both bradykinin and kallidin elicited a concentration-dependent (1-100 nM) depolarisation when applied to the superfusate bathing the nodose ganglia, whereas the bradykinin B1 receptor agonist, des-Arg9-bradykinin, was only effective in the micromolar range. Furthermore, the electrophysiological response to bradykinin was antagonised by the bradykinin B2 receptor antagonist, D-arginyl-L-arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-t hienyl)-L-alanyl-L-seryl-D-1,2,3,4-tetrahydro-3-isoquinolinecarbonyl+ ++-L-(2alpha,3beta,7abeta)-octahydro-1H-indole-2-carbonyl-L- arginine (Hoe 140), in a concentration-related manner. To determine the anatomical location of functional bradykinin B2 receptors, in vitro autoradiography with [125I]para-iodophenyl Hoe 140 was performed on sections of rat and human inferior vagal (nodose) ganglia and confirmed the presence of binding over vagal perikarya. Collectively, these data provide evidence for functionally relevant bradykinin B2 receptors on vagal afferent neurons, which are apparently also present on human vagal perikarya.

Adrenergic beta-Antagonists↗

PubMed1998

Effects of acute ethanol on GABA release and GABA(A) receptor density in the rat mesolimbic system.

The present study has addressed whether acute ethanol administration mediates changes in GABA release and GABA(A) receptor density in the rat mesolimbic system. In vivo microdialysis was performed in the ventral pallidum and between the ventral tegmental area and substantia nigra of conscious ethanol-naive rats. Extracellular levels of endogenous GABA were stable in both brain regions and not significantly affected following administration of 5 ml of 20% ethanol by gavage, despite clear overt behavioral signs of intoxication. Two hours following ethanol administration, animals were decapitated and the brains processed for autoradiography. Adjacent tissue sections were incubated with [3H]SR95531 or [3H]muscimol and the resulting autoradiograms quantified. Binding of both radioligands was significantly reduced in the striatum of rats treated with ethanol compared to vehicle (15 +/- 2% for [3H]SR95531 and 33 +/- 6% for [3H]muscimol). In contrast, ethanol had no effect on [3H]SR95531 binding in hippocampus, while that of [3H]muscimol was increased by 100 +/- 30%. Ethanol had no effect on the binding of either radioligand in all other areas examined. Therefore, while acute ethanol had no effect on the release of GABA in either the ventral pallidum or ventral tegmental area, changes in the binding density of GABA(A) ligands were observed in selected brain regions.

Administration, Oral↗

PubMed1998

In vivo microdialysis reveals facilitatory metabotropic glutamate receptors regulating excitatory amino acid release in rat nucleus tractus solitarius.

The functional involvement of metabotropic glutamate receptors in nucleus tractus solitarius of the medulla has been examined by evaluating the action of the selective metabotropic glutamate receptor agonist trans-1-amino-cyclopentane-1,3-dicarboxylate on the extracellular release of L-glutamate and L-aspartate by in vivo microdialysis. Studies were performed in urethane anaesthetized rats, with a microdialysis probe being inserted into the medial nucleus tractus solitarius, which was constantly perfused with artificial cerebrospinal fluid (1 microliter/min). Following an equilibration period, samples were collected for amino acid analysis by HPLC. trans-1-Amino-cyclopentane-1,3-dicarboxylate (30 microM) produced 190% and 500% increases in basal release of L-glutamate and L-aspartate, respectively. trans-1-Amino-cyclopentane-1,3-dicarboxylate-evoked release of L-glutamate and L-aspartate was concentration-dependent (10-100 microM), and was calcium and tetrodotoxin-sensitive. Two metabotropic glutamate receptor antagonists, (+)-alpha-methyl-4-carboxyphenylglycine (200 microM) and (S)-4-carboxyphenylglycine (500 microM), when administered via a microdialysis probe, significantly attenuated the trans-1-amino-cyclopentane-1,3-dicarboxylate-evoked release of both L-glutamate and L-aspartate. Basal release was not altered by these two antagonists. Histological studies verified the location of the probe in the nucleus tractus solitarius. These studies provide the first evidence using in vivo microdialysis of functional metabotropic glutamate receptors, which seem to be facilitatory pre- and postsynaptically located receptors, in the rat nucleus tractus solitarius. On the basis of our findings, metabotropic glutamate receptors are likely to play key modulatory roles in regulating transmitter release.

Animals↗