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Fine distribution of gamma-aminobutyric acid, glutamic acid decarboxylase, and glutamic acid in the rabbit cerebellum.

The fine distribution of GABA, glutamic acid decarboxylase, and glutamic acid within each layer of the rabbit cerebellar cortex was determined with microanalytical methods. The greatest glutamic acid decarboxylase activity and the highest GABA concentration were found in the Purkinje cell layer. In the distribution of GABA and glutamic acid decarboxylase the peak of glutamic acid decarboxylase activity was more pronounced than that of GABA; the concentration of glutamic acid did not show much variation between each layer.

Animals

A novel prodrug of salicylic acid, salicylic acid-glutamic acid conjugate utilizing hydrolysis in rabbit intestinal microorganisms.

The fate of salicylic acid-glutamic acid conjugate (salicyl-glutamic acid) following oral, intravenous, intracecal and rectal administration (60, 10, 5 and 5 mg/kg, respectively: salicylic acid equivalent) was examined in rabbits. Salicylic acid was detected in the blood 2 h after oral administration of salicyl-glutamic acid and it reached the maximum level (69.4 micrograms/ml) at 18 h after the dose. A high blood concentration of salicylic acid (24.8 micrograms/ml) was observed up to 36 h. But only a small amount of salicyl-glutamic acid was detected in the blood (less than 2.5 micrograms/ml, as salicylic acid). In contrast, unchanged salicyl-glutamic acid was found mainly in the blood following intravenous administration of salicyl-glutamic acid, suggesting that presystemic de-conjugation of salicyl-glutamic acid predominantly occurred. The intestinal mucosal de-conjugation of salicyl-glutamic acid was negligible in the in situ intestinal sac preparation with complete mesenteric venous blood collection. Immediate and very extensive salicylic acid formation in the cecum was found following intracecal administration of salicyl-glutamic acid. After oral pretreatment of rabbits with kanamycin sulfate (6 x 400 mg), a significant inhibition of salicylic acid formation following intracecal administration of salicyl-glutamic acid was observed, indicating that the intestinal microorganisms were responsible for the biotransformation of salicyl-glutamic acid. Also, in vitro incubation of salicyl-glutamic acid with gut contents showed that the primary location of hydrolysis was the hind gut.

Animals

Effect of transient neonatal hypothyroidism on the free aspartic acid, glutamic acid, and GABA content of different central auditory regions in the rat.

The aspartic acid, glutamic acid, and gamma-aminobutyric acid (GABA) contents were determined in four central auditory system regions in rats with transient neonatal hypothyroidism compared with control ones: the ventral and dorsal parts of the cochlear nucleus, the central nucleus of the inferior colliculus, the auditory cortex, and in an extra-auditory structure, the substantia nigra pars reticulata. The animals were sacrificed at 50 days of age, brain tissue samples were taken out by microdissection, and the free amino acids were extracted. The amino acid content was assessed by double-isotope labelling following two-dimensional thin-layer chromatography separation. GABA content was significantly decreased in both cochlear nucleus regions and glutamic acid was elevated in the inferior colliculus. Neonatal hypothyroidism had no significant effect on the aspartic acid levels in the regions studied. The results suggest an effect of neonatal hypothyroidism on regional contents of free amino acids known as candidate neurotransmitters in the auditory system.

Animals

A peptide containing aspartic acid, glutamic acid and serine in calf brain synaptic vesicles.

Free amino acids and other amino compounds in calf brain synaptic vesicles were identified and determined by thin-layer chromatography and ion-exchange chromatography. The vesicles contained ten identified amino acids with glutamic acid, aspartic acid, taurine and gamma-aminobutyric acid in the highest concentrations, and also cysteic acid (or cysteinesulfinic acid), glutamine, alanine, serine, glycine and lysine. The vesicles also contained certain unknown acid-labile, ninhydrin-positive compounds, one of which was a peptide yielding, after acid hydrolysis, about 40% aspartic acid, 30% serine, 15% glutamic acid, 10% glycine and possibly some alanine and lysine. The concentration of the peptide in the vesicles was as high as that of all the other amino compounds together.

Amino Acids

Discrimination of Rhizobium japonicum, Rhizobium lupini, Rhizobium trifolii, Rhizobium leguminosarum and of bacteroids by uptake of 2-ketoglutaric acid, glutamic acid and phosphate.

Rhizobium strains (one each of Rh. japonicum, Rh. lupini, Rh. leguminosarum) take up 2-ketoglutaric acid in general much faster and from lower concentrations in the medium than strains of Escherichia coli, Bacillus subtilis and Chromobacterium violaceum. A strain of Enterobacter aerogenes, however, is more similar to some Rhizobium strains. The same strains of Rhizobium take up also phosphate much faster and from lower concentrations than the other bacteria tested. 4 strains of Rh. lupini proved to be significantly different from 4 strains of Rh. trifolii in taking up L-glutamic acid from three to ten times lower concentration within 5 h. A similar difference was noticed between 5 strains of Rh. leguminosarum and 2 strains of Rh. japonicum for the uptake of 2-ketoglutaric acid and of L-glutamic acid. Isolated bacteroids from nodules of Glycine max var. Chippeway have a reduced uptake capacity for glutamic acid and for 2-ketoglutaric acid during the first 10-12 h, but reach the same value after 24 h as free living Rh. japonicum cells. The differences in the uptake kinetics are independent of cell concentration. The group II Rhizobium strains (Rh. japonicum and Rh. lupini, slow growing Rhizobium) are characterized by a rapid uptake of glutamic acid to a low remaining concentration of 1-3 X 10(-7) M and an uptake of 2-ketoglutaric acid to a remaining concentration of 2-5 X 10(-7) M. The group I Rhizobium strains (Rh. trifolii and Rh. leguminosarum, fast growing Rhizobium), can be characterized by a much slower uptake of both substances with a more than ten times higher concentration of both metabolites remaining in the medium after the same time.

Bacillus subtilis

[Age estimation by amino acid racemization in teeth. A comparison of data for aspartic acid, glutamic acid and alanine].

On the age estimation by the amino acid racemization analysis of dentin, besides the utilization of aspartic acid (Asp) as described in earlier reports, we further studied relationships between the D/L ratios based on glutamic acid (Glu) as well as alanine (Ala) and actual ages. The study was followed up by comparing racemization velocities of the three amino acids under some heating experiments. At four steps (6, 24, 48 and 72 hours) of hydrolysis, the coefficient values of D/L ratio of each amino acid and actual age were calculated as 0.986 to 0.994 for Asp, 0.522 to 0.806 for Glu, and 0.577 to 0.737 for Ala. The data indicate that Asp gives an extremely good result. Glu and Ala do provide reliable D/L ratios, however they are not in proportion to actual ages. Consequently, Glu and Ala seem to be much less suitable for utilization in age estimation. Reaction rate constants (k.yr-1) of racemization of Asp, Glu and Ala in antemortem teeth were 5.3825 x 10(-4), 5.1000 x 10(-5) and 2.3875 x 10(-5), respectively. Those in teeth left drying at 15 degrees C were 2.4850 x 10(-8), 1.9119 x 10(-9), and 1.11450 x 10(-9), respectively. Assuming that the reaction velocity of Asp be 1 in both living and dry states, that of Glu were calculated as 0.09 and 0.08, that of Ala, 0.04 and 0.05, indicating very similar rates. The result confirmed that both Glu and Ala gave considerably slow racemization velocities as compared with Asp.

Age Determination by Teeth

Gamma-aminobutyric acid, glutamic acid decarboxylase and tyrosine hydroxylase in rat striatum demonstrated by single and dual immunocytochemistry.

By means of dual ultrastructural immunostaining the followings patterns are visualized: gamma-aminobutyric acid (GABA) immunoreactive neurons, dendrites, axons and axon terminals and tyrosine hydroxylase (TH) immunopositive fibers, varicosities and boutons in rat striatum. Additionally single glutamic acid decarboxylase (GAD) immunolabeling is carried out. Four subgroups of GABA and GAD immunoreactive striatal neurons are revealed. These neuronal types are identified on the basis of sectional diameters, nuclear form and nuclear envelope invaginations, quantity of cytoplasm and cell organelles. Plasmalemmal appositions between GABAergic and between GABAergic and immunonegative neurons are observed. All subgroups of striatal GABAergic neurons contact with GABA and GAD immunoreactive, TH immunoreactive and immunonegative boutons. In the striatal neuropil numerous GABAergic, dopaminergic and immunonegative axonal endings synapsed with dendrites and spines are found out. Massive dopaminergic striatal structures using dual immunostaining is evident. Some GABA and GAD immunoreactive dendrites are revealed in direct contact with capillary walls.

Animals

Serotonin-stimulated release of [3H]dopamine via reversal of the dopamine transporter in rat striatum and nucleus accumbens: a comparison with release elicited by potassium, N-methyl-D-aspartic acid, glutamic acid and D-amphetamine.

Release of preloaded radiolabeled dopamine ([3H]DA) elicited by several agents from terminal fields of mesolimbic and nigrostriatal projections in rats was compared. Several similarities between the two areas were observed. For example, potassium, which stimulates release both directly, through altering the potential across the membrane of the dopaminergic neuron, as well as indirectly, presumably by releasing endogenous excitatory neurotransmitters, exhibited some similarities to release stimulated by L-glutamate and N-methyl-D-aspartic acid. These included sensitivity to tetrodotoxin (TTX), Mg++ and Ca++. In contrast, release of [3H]DA stimulated by serotonin (5-HT), like that stimulated by D-amphetamine, depended upon a functional dopamine transport system and was less sensitive to TTX, Mg++ and Ca++. 5-HT-stimulated [3H]DA release in striatum (STR) and nucleus accumbens (NACC) was not modified by antagonists at 5-HT2 or 5-HT3 receptors. Differences were observed in release of [3H]DA from STR and NACC. Elevated potassium (20 mM) released about twice as much [3H]DA from NACC as it did from STR. 5-HT was also able to release more [3H]DA from NACC than from STR. Conversely, D-amphetamine released more [3H]DA from STR than from NACC. TTX increased release stimulated by potassium in STR, but decreased release stimulated by potassium in NACC. These observations suggest that receptor- and non-receptor-mediated mechanisms may contribute to regulation of [3H]DA release in mesolimbic and nigrostriatal areas of the brain. It is possible that endogenous 5-HT in STR or NACC acts as a local regulator of DA release acting via a transport-dependent mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)

Amphetamine

Glutamic acid and glutamic acid diethylester in tinnitus treatment.

Numerous recent findings indicate that in mammals glutamic acid (Glu) functions as the primary and secondary afferent cochlear transmitter, or at least as an agonist of the main transmitter. Glutamic acid diethylester (GDEE) is one of the known antagonists of Glu. A long term suppression of certain forms of tinnitus was observed dependent on the sequence, amount and perfusion rate upon i.v. application of Glu and GDEE. Consecutive controls with tinnitus detecting as well as blind studies helped to objectify the subjective sensations of the patients.

Glutamates