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gamma-Aminobutyric acid and taurine release in the striatum of the rat during hypoglycemic coma, studied by microdialysis.

Extracellular levels of striatal gamma-aminobutyric acid (GABA) and taurine were monitored during insulin-induced hypoglycemia using microdialysis. At the onset of isoelectricity in the electroencephalogram (EEG), a transient 5-fold increase in the levels of GABA occurred. Taurine levels increased 5 min following the onset of isoelectricity and continued to increase during the entire isoelectric period. The results demonstrate that events associated with the onset of isoelectricity during hypoglycemia trigger an increase in extracellular concentrations of GABA and taurine. The discrepancy in time-course of these changes may reflect differences in compartmentation, function and metabolism of the two amino acids.

Animals↗

Gamma-aminobutyric acid is present in a spatially discrete subpopulation of hair cells in the crista ampullaris of the toadfish Opsanus tau.

Although gamma-aminobutyric acid (GABA) and glutamate are known to be present in the vestibular sensory epithelia of a variety of species, the functional relationship between these two transmitters is not clear. The present study addresses the three-dimensional spatial distribution of GABA and glutamate immunoreactivity in the vestibular labyrinth of the oyster toadfish by using whole end organs labeled by immunofluorescence with monoclonal anti-GABA and/or antiglutamate antibodies and visualized as whole mounts by multiphoton confocal microscopy. We find glutamate-immunoreactive hair cells present throughout the sensory epithelium. In contrast, prominent GABA immunoreactivity is restricted to a small population of hair cells located in the central region of the crista. Double immunofluorescence reveals two distinct staining patterns in GABA-labeled hair cells. Most ( approximately 80%) GABA-labeled cells show trace levels of glutamate, appropriate for the metabolic/synthetic role of cytoplasmic glutamate. The remainder of the GABA-stained cells contain substantial levels of both GABA and glutamate, suggesting transmitter colocalization. In the toadfish utricle, glutamatergic hair cells are present throughout the macula. GABA-immunoreactive hair cells follow the arc of the striola, and most GABA-labeled receptor cells coexpress glutamate. The localization of GABA was explored in other species as well. In the pigeon, GABAergic hair cells are present throughout the crista ampullaris. Our findings demonstrate that multiple, neurochemically distinct types of hair cells are present in vestibular sensory epithelia. These observations, together with the excitatory activity generally associated with 8th nerve afferent fibers, strongly suggest that GABA serves an important, specific, and complex role in determining primary afferent response dynamics.

Animals↗

gamma-Aminobutyric acid uptake by rat kidney brush-border membrane vesicles.

Brush-border membrane vesicles (BBMV) from rat kidney cortex possessed two uptake systems for gamma-aminobutyric acid (GABA), a high affinity system (Km = 10.9 microM) and a low affinity system (Km = 1203 microM). Both uptake systems were inhibited by p-hydroxymercuribenzoic acid and ouabain, and by the action of neuraminidase, whereas the GABA analogs nipecotic acid, beta-alanine, 2,4-diaminobutyric acid and 4,5,6,7-tetrahydroisoxazolo-[4,5 c]-pyridin-3-ol had no effect on the GABA uptake activity. The BBMV uptake systems were clearly different from the GABA transport systems present in brain tissue.

Animals↗

gamma-Aminobutyric acid (GABA) uptake systems in human frontal cortex.

Synaptosomes prepared from human frontal cortex possessed two uptake systems for gamma-aminobutyric acid (GABA) with Km values of 1.9 and 20.6 microM respectively, and with Vmax values of 0.15 and 0.78 nmol/min/mg protein respectively. In this respect, human tissue preparations differed from cortical synaptosomes from rodents in that a third, low-affinity GABA uptake system was not detectable in human synaptosomes.

Aged↗

Effects of gamma-aminobutyric acid on the compound action potential of the rat superior cervical ganglion.

The effects on the ganglionic transmission of gamma-aminobutyric acid (GABA) and related drugs were studied in the rat isolated superior cervical ganglion. The extracellularly recorded postganglionic compound action potential was used as an index for ganglionic transmission. GABA reversibly inhibited the ganglionic transmission by blocking the action potential. The effect of GABA is markedly antagonized by bicuculline (20 microM) or picrotoxin (200 microM). Furosemide (1 mM) was also effective in antagonizing the ganglionic effect of GABA. The antagonism by bicuculline was not affected in the presence of pentobarbital (50 microM). Baclofen inhibited the ganglionic transmission through an activation of GABAB receptors since it was markedly antagonized by phaclofen. Muscimol and isoguvacine were less effective than GABA in blocking the ganglionic transmission. The GABA uptake inhibitor isonipecotic acid blocked the ganglionic transmission, probably by activating GABAA receptors, because the effect was antagonized by bicuculline, but the GABA uptake inhibitor guvacine had no significant effect on the ganglionic transmission. These results suggest that GABA causes a ganglionic blockade by activation of both GABAA and GABAB receptor subtypes.

Action Potentials↗

Glutamine stimulates gamma-aminobutyric acid synthesis in synaptosomes but other putative astrocyte-to-neuron shuttle substrates do not.

GABAergic neurons require a supply of precursor glutamate for gamma-aminobutyric acid (GABA) synthesis to maintain their GABA levels. Because neurons lack the anaplerotic enzymes necessary for net synthesis of glutamate from glucose, they depend on astrocytes to supply compounds that can be metabolized to glutamate and ultimately used for GABA production. To test the effect of putative astrocytic shuttle metabolites on GABA synthesis, we used synaptosomes prepared from substantia nigra, an area rich in GABAergic terminals. The low number of glutamatergic endings in the nigral preparation allows a more accurate measurement of glutamate present in GABAergic endings. GABA synthesis by nigral synaptosomes was stimulated 3.1-fold when 500 microM glutamine was added to the incubation medium. Glutamate amounts also increased. In contrast, the possible precursor metabolites. 2-oxoglutarate (2-OG), malate and citrate, failed to stimulate GABA synthesis over the rate observed with control medium. Unlike malate and citrate. 2-OG reduced the decline in total glutamate observed when synaptosomes were incubated in control. In contrast to glutamine the production of synaptosomal glutamate from 2-OG, malate, and citrate is not great enough to stimulate GABA synthesis.

Amino Acids↗

Cardiovascular effects of L-glutamate and gamma-aminobutyric acid injected into the rostral ventrolateral medulla in normotensive and spontaneously hypertensive rats.

We studied the responses to chemical stimulation of the ventrolateral medulla in pentobarbital-anaesthetized, paralyzed, normotensive and spontaneously hypertensive (SHR) rats. When unilaterally injected into a circumscribed region of the rostral ventrolateral medulla, L-glutamate (0.16-1.6 nmol) elicited a dose-dependent increase in arterial pressure and heart rate. Bilateral microinjections of L-glutamate diethylester (63 nmol), an antagonist of excitatory amino acids, into the glutamate-sensitive sites markedly reduced arterial pressure and heart rate. gamma-Aminobutyric acid (GABA) (0.3-3 nmol) injected into the glutamate-sensitive sites caused a dose-dependent decrease in arterial pressure and heart rate. The depressor response to GABA was smaller in SHR than that in normotensive Wistar Kyoto rats (WKY), while there were no differences between WKY and SHR in the pressor response to L-glutamate. Thus, a depressor function involving the ventrolateral medulla appears to be diminished in SHR.

Animals↗

A partial structure for the gamma-aminobutyric acid (GABAA) receptor is derived from the model for the nicotinic acetylcholine receptor. The anion-exchange protein of cell membranes is related to the GABAA receptor.

Based on the nicotinic acetylcholine receptor model [(1987) Eur. J. Biochem. 168, 431-449], a partial model is constructed for the exobilayer portion of the GABAA receptor, an approach justified by the superfamily relationship of the two receptors [(1987) Nature 328, 221-227]. The model predicts successfully the excess positive charge on interior strands which constitute the ligand-responsive portion of the receptor. Binding to GABA expands the exobilayer portion of the receptor, opening a pathway to a chloride channel. Separate binding sites for antianxiolytics (benzodiazepines) and hypnotics (barbiturates) are suggested, with prolongation of chloride entry projected as a consequence of stabilization of the open form. The anion-exchange protein (AEP) of membranes (band 3 of red blood cell membranes) is similar in some respects to the gamma-aminobutyric acid (GABAA) receptor. Both proteins are inhibited and labeled by diisocyanatostilbenedisulfonate (DIDS), both transport Cl- and HCO-3, and both are membrane proteins. Starting with the lysines known to be labeled in band 3 protein, searches of the amino acid sequences of the GABAA receptor alpha- and beta-subunits reveal at least 4 reasonably homologous sequences. The relationship between AEP and GABAA receptor leads to the idea that the chloride/bicarbonate channel may be the ancestor of all ligand-gated channels, with ligand gating by gamma-aminobutyric acid and acetylcholine arising later in evolution.

Amino Acid Sequence↗

Synaptically released glutamate reduces gamma-aminobutyric acid (GABA)ergic inhibition in the hippocampus via kainate receptors.

Exogenous application of agonists at the kainate subtype of glutamate receptors has been shown to depress evoked monosynaptic inhibition by gamma-aminobutyric acid (GABA)ergic interneurons in the hippocampus. This observation has led to the hypothesis that synaptic release of endogenous glutamate might have a disinhibitory effect on neuronal circuits, in addition to depolarizing neurons via postsynaptic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), kainate, and N-methyl-D-aspartic acid (NMDA) receptors. It is not known, however, if glutamate released from excitatory neurons has the same kainate receptor-mediated effect on monosynaptic inhibitory transmission as exogenous agonist application. Indeed, the recent demonstration that excitatory synaptic signals elicited in interneurons are partly mediated by kainate receptors suggests that these receptors may have a pro- rather than disinhibitory role. Here, we examine the effect of synaptically released glutamate on monosynaptic inhibitory signaling. In the presence of antagonists to AMPA and NMDA receptors, brief bursts of activity in glutamatergic afferent fibers reduce GABAergic transmission. This depression of inhibition is reversibly abolished by blocking kainate receptors. It persists when GABA(B) receptors are blocked and is enhanced by blocking metabotropic glutamate receptors, possibly explained by presynaptic regulation of glutamate release from excitatory afferents by metabotropic autoreceptors. We conclude that the net kainate receptor-mediated effect of synaptically released glutamate is to reduce monosynaptic inhibition. Since this form of disinhibition may contribute to seizure initiation, kainate receptors may constitute an important target for anticonvulsant drug development.

2-Amino-5-phosphonovalerate↗

Expression of gamma-aminobutyric acid and calcium binding protein-parvalbumin by chick motoneurons.

The expression of calcium binding protein parvalbumin (PV) and gamma-aminobutyric acid (GABA) was studied in the chick motoneurons by using pre- and postembedding immunocytochemistry. Our data reveal that PV and GABA are colocalized in the majority, but not all, of chick lumbo-sacral spinal motoneurons innervating the somatic muscles. It is suggested that, in this neuromuscular system, GABA does not act as a classical inhibitory neurotransmitter but, combined with calcium, could be involved, at least in part, in the maintenance of neurons and the prevention of cell death as in certain neurodegenerative disorders.

Animals↗

An arylaminopyridazine derivative of gamma-aminobutyric acid (GABA) is a selective and competitive antagonist at the GABAA receptor site.

In view of finding a new gamma-aminobutyric acid (GABA) receptor ligand we synthesized an arylaminopyridazine derivative of GABA, SR 95103 [2-(carboxy-3'-propyl)-3-amino-4-methyl-6-phenylpyridazinium chloride]. SR 95103 displaced [3H]GABA from rat brain membranes with an apparent Ki of 2.2 microM and a Hill number near 1.0. SR 95103 (1-100 microM) antagonized the GABA-mediated enhancement of [3H]diazepam binding in a concentration-dependent manner without affecting [3H]diazepam binding per se. SR 95103 competitively antagonized GABA-induced membrane depolarization in rat spinal ganglia. In all these experiments, the potency of SR 95103 was close to that of bicuculline. SR 95103 (100 microM) did not interact with a variety of central receptors--in particular the GABAB, the strychnine, and the glutamate receptors--did not inhibit Na+-dependent synaptosomal GABA uptake, and did not affect GABA-transaminase and glutamic acid decarboxylase activities. Intraperitoneally administered SR 95103 elicited clonicotonic seizures in mice (ED50 = 180 mg/kg). On the basis of these results it is postulated that St 95103 is a competitive antagonist of GABA at the GABAA receptor site. In addition to being an interesting lead structure for the search of GABA ligands, SR 95103 could also be a useful tool to investigate GABA receptor subtypes because it is freely soluble in water and chemically stable.

Animals↗

[The influence of rabies immunization of gamma-aminobutyric acid metabolism in the brains of animals].

Subcutaneous injection to albino rats (100-120 g) of lived fixed rabies virus was accompanied by a brief marked decrease in the content of gamma-aminobutryic acid in the animals' brain. There was also an increase in the activity of gamma-aminobutyric acid alpha-ketoglutaric acid transaminase in the brain tissue of animals vaccinated with live fixed rabies virus.

4-Aminobutyrate Transaminase↗

Detection of gamma-aminobutyric acid (GABA) by longitudinal scalar order difference editing.

Two novel spectral editing techniques for the in vivo detection of gamma-aminobutyric acid (GABA) are presented. The techniques rely on the generation of longitudinal scalar order (LSO) coherences, which in combination with J-difference editing results in the selective detection of GABA. The utilization of LSO coherences makes the editing sequences insensitive to phase and frequency instabilities. Furthermore, the spectral editing selectivity can be increased independent of the echo time, thereby opening the echo time for state-of-the-art water suppression and/or spatial localization techniques. The performance of the LSO editing techniques is theoretically demonstrated with product operator calculations and density matrix simulations and experimentally evaluated on phantoms in vitro and on human brain in vivo.

Brain↗

A role for protein kinase C in the electrically evoked release of [3H] gamma-aminobutyric acid in rabbit caudate nucleus.

A possible participation of protein kinase C (PKC) in depolarization-induced release of gamma-aminobutyric acid (GABA) in rabbit caudate nucleus was examined by means of phorbol esters and staurosporine. Slices of caudate nucleus were loaded with [3H]GABA, then superfused and stimulated electrically (3 ms, 5 Hz, 24 mA, 5 V/cm) for 2 min. Aminooxyacetic acid and the uptake inhibitor nipecotic acid were present throughout. The PKC activator 4 beta-phorbol 12,13-dibutyrate (4 beta-PDB) markedly enhanced the evoked [3H]GABA release. In contrast, its biologically inactive isomer, 4 alpha-PDB, did not facilitate transmitter release. Staurosporine, an inhibitor of PKC, diminished [3H]GABA release and counteracted the effects caused by 4 beta-PDB. The above results suggest a participation of PKC in depolarization-induced GABA release in rabbit caudate nucleus. The mechanism underlying the modulation of GABA release by PKC seems to be independent of presynaptic GABA, dopamine and 5-hydroxytryptamine receptors.

Alkaloids↗

[The effect of pain caused by multiple trauma on gamma-aminobutyric acid metabolism and the functional condition of animals].

Multiple injury to the bone tissue in rats caused changes of gamma-aminobutyric acid (GABA) metabolism manifested first by activation of its synthesis and then by inhibition of enzymatic inactivation and increase in GABA content in the brain. The initial reaction of the GABA system was attended by a loss of total body and thymus weight, increase in adrenal weight, and bicocculin- and picrotoxin-sensitive hypoalgesia in the tail-flick test. The subsequent changes of GABA-ergic transmission developed during restoration of the animals' functional condition but body weight loss persisted. It is suggested that activation of the GABA system is a defence reaction tending to intensify the inhibition processes and increase the activity of the brain antinociceptive systems.

Animals↗