Synthesis and release of amino acid transmitters.
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Animals↗
Biomedical subjects
F Fonnum
Publications and source records attributed to F Fonnum.
Prenatal effects of trichlorfon on the guinea pig brain.
The organophosphorus compound trichlorfon, dimethyl (2,2,2-trichloro-1-hydroxyethyl) phosphonate, was administered by stomach tube (100 mg/kg) to pregnant guinea pigs at two different stages of gestation (days 36, 37, 38 and 51, 52, 53). The pups developed locomotory disturbances, and post mortem examination revealed significantly decreased weights of the total brain and the cerebellum, as compared to controls. There was also a significant weight reduction particularly of the medulla oblongata, but also of the hippocampus, the thalamus, and the colliculi. Histological examination of the cerebellum revealed reduction of the external granular layer and the molecular layer, and regional absence of Purkinje cells. The activities of the neurotransmitter enzymes choline acetyltransferase (ChAT), and glutamate decarboxylase (GAD) in cerebellum were reduced as compared to the control values.
Animals↗
Differences in anionic dependence of the synaptic efflux of D-aspartic acid and gamma-aminobutyric acid.
The synaptosomal effluxes of D-aspartate and gamma-aminobutyric acid (GABA) induced by a substitution of the Cl- ions with propionate in the incubation medium were measured in preparations of hippocampal tissue homogenates. The efflux of aspartate was significantly higher than spontaneous efflux at 125 mM Cl- (normal = 144 mM) and was increased with decreasing Cl- concentration. GABA efflux was much less sensitive to a reduction in Cl- concentration than D-aspartate. The efflux was Ca2+-dependent in both cases.
Animals↗
Effect of insulin-induced hypoglycemia on the concentrations of glutamate and related amino acids and energy metabolites in the intact and decorticated rat neostriatum.
The glutamate (Glu) terminals in rat neostriatum were removed by a unilateral frontal decortication. One to two weeks later the effects of insulin-induced hypoglycemia on the steady-state levels of amino acids [Glu, glutamine (Gln), aspartate (Asp), gamma-aminobutyric acid (GABA), taurine] and energy metabolites (glucose, glycogen, alpha-ketoglutarate, pyruvate, lactate, ATP, ADP, AMP, phosphocreatine) were examined in the intact and decorticated neostriatum from brains frozen in situ. The changes in the metabolite levels were examined during normoglycemia, hypoglycemia with burst-suppression (BS) EEG, after 5 and 30 min of hypoglycemic coma with isoelectric EEG, and 1 h of recovery following 30 min of isoelectric EEG. In normoglycemia Glu decreased and Gln and glycogen increased significantly on the decorticated side. During the BS period no significant differences in the measured compounds were noted between the two sides. After 5 min of isoelectric EEG Glu, Gln, GABA, and ATP levels were significantly lower and Asp higher on the intact than on the decorticated side. No differences between the two sides were found after 30 min of isoelectric EEG. After 1 h of recovery from 30 min of isoelectric EEG Glu, Gln, and glycogen had not reached their control levels. Glu was significantly lower, and Gln and glycogen higher on the decorticated side. The Asp and GABA levels were not significantly different from control levels. The results indicate that the turnover of Glu is higher in the intact than in decorticated neostriatum during profound hypoglycemia.
Amino Acids↗
A bioluminescence method for the measurement of L-glutamate: applications to the study of changes in the release of L-glutamate from lateral geniculate nucleus and superior colliculus after visual cortex ablation in rats.
We have developed a rapid, simple, specific, and very sensitive bioluminescence method for the measurement of L-glutamate (L-Glu). Oxidation of L-Glu by glutamate dehydrogenase has been coupled with bacterial FMN reductase and luciferase. Light production (i.e., peak height or integral) was linear from less than 0.5 to 500 pmol of L-Glu. Potential interfering substances that may be encountered in brain tissue have been identified. The most potent inhibitors were ascorbate and the biogenic amines. Procedures that conferred long-term stability of the reagent mixture (greater than 8 h) were established. Bioluminescence analysis of L-Glu content in brain tissue extracts, fractions from release experiments, and human CSF corroborated respective results obtained by HPLC analysis. In this study, we have applied the method to monitor changes in the KCl-evoked release of endogenous L-Glu from milligram amounts of brain tissue, i.e., from lateral geniculate nucleus and superior colliculus after visual cortex ablation.
Animals↗
Presynaptic inhibition of acetylcholine release.
High potassium (51 mM) has been shown to evoke release of acetylcholine ([3H]ACh and endogenous ACh) from cholinergic nerves in rat bronchial smooth muscle. The release of [3H]ACh was reduced by 85% when the Ca2+ concentration was changed from 2 to 0.1 mM. The veratridine-induced release was completely inhibited by tetrodotoxin, but tetrodotoxin did not reduce the potassium-evoked release. The muscarinic agonist, oxotremorine, reduced the potassium stimulated release of [3H]ACh, without affecting the basal release. In contrast, scopolamine substantially potentiated the potassium-evoked release. Adenosine had a dual effect in the rat bronchi. Adenosine inhibited the potassium-evoked release of [3H]ACh and this presynaptic effect of adenosine was antagonized by 8-phenyltheophylline. Adenosine also induced contraction of the bronchial smooth muscle and there was potentiation by adenosine of the ACh-induced contraction. The results indicate that cholinergic nerve terminals in the rat bronchi possess muscarinic receptors which inhibit the release of ACh. Adenosine may have analogous effects, e.g. presynaptic inhibition of transmitter release in addition to postsynaptic enhancement of bronchial smooth muscle contraction.
Acetylcholine↗
The effect of trimethyltin on three glutamergic and gabaergic transmitter parameters in vitro: high affinity uptake, release and receptor binding.
The effects of trimethyltin (TMT) on high-affinity uptake, release and sodium-independent binding of glutamic acid and gamma-aminobutyric acid (GABA) were studied in vitro in homogenates of hippocampal tissue. TMT (50 micron) increased the release of glutamic acid from synaptosomes in the resting state (5 mM K+), whereas the release of GABA was only slightly affected. High affinity uptake of glutamate was inhibited by TMT in the same concentration range as release. The uptake of GABA was only affected by TMT-concentrations from 500 micron to 5 mM. The sodium independent binding of both glutamate and GABA, usually assumed to be binding to receptor sites, were inhibited with 50 microM or more TMT in the incubation medium. The results indicate that TMT can interfere with several different events of the neurotransmission process in the central nervous system at concentrations which can be obtained in the brain of rats after a sublethal dose of the compound.
Animals↗
Elevated concentrations of glutamate and aspartate in human ventricular cerebrospinal fluid (vCSF) during episodes of increased CSF pressure and clinical signs of impaired brain circulation.
In the ventricular cerebrospinal fluid (vCSF) of 10 hydrocephalic patients the mean (+/- S.D.) concentrations of glutamate and asparate were 2.9 +/- 0.2 and 0.2 +/- 0.2 microM, respectively. Significantly higher concentrations of these amino acids were found in two patients (glutamate 37.8 and 22.4 microM, aspartate 2.2 and 0.6 microM) with symptoms of impaired brain tissue perfusion, i.e. relative ischemia due to severely increased intraventricular CSF pressure. Our results are consistent with recent experiments in rats showing increased extracellular concentrations of glutamate and aspartate during transient cerebral ischemia.
Adult↗
A method for generating toxic vapors of soman: toxicity of soman by inhalation in rats.
A method for administration of highly toxic chemicals by inhalation was developed. The model has three features of special interest: (1) a diffusion cell for producing a constant gas concentration, if necessary for several hours and days, (2) a small rapidly equilibrated inhalation chamber (1100 ml), and (3) complete isolation of the toxic chemicals from the atmosphere. The LCt50 of the anticholinesterase soman [o-(1,2,2 trimethylpropyl)-methyl-phosphonofluoridate] was 400 mg min/m3, registered 24 hr after the end of exposure. The lethal concentration X time of soman was 520 +/- 60 mg min/m3 when exposing the animals until death in the inhalation chamber. The exposure was less than 30 min and the concentration of soman was 21 mg/m3. The inhibition of acetylcholinesterase, cholinesterase, and carboxylesterase activities in different tissues was analyzed to study the possible barrier mechanisms that might exist in the body to soman. There was a large inhibition of the carboxylesterase and cholinesterase activities in bronchi and lungs as well as in blood. Carboxylesterases were important as detoxifying enzymes, as shown by 70% enhancement in toxicity of soman following sc pretreatment with TOCP (tri-ortho-cresyl-phosphate), a carboxylesterase inhibitor.
Acetylcholinesterase↗
A radiochemical assay method for carboxylesterase, and comparison of enzyme activity towards the substrates methyl [1-14C] butyrate and 4-nitrophenyl butyrate.
A radiochemical assay for carboxylesterase based on the substrate methyl[1-14C]butyrate is described. The blank value corresponds to 1.04 micrograms (liver)-1.44 mg (plasma) of tissues with the highest and lowest activity respectively, which constitute the sensitivity of the method. The hydrolysis of methyl butyrate and 4-nitrophenyl butyrate by plasma, liver, lung, heart, diaphragm, cerebrum, kidney and duodenum of rat have been compared. The results showed that the two substrates were hydrolysed preferentially by two different groups of the enzyme. The effect of selective esterase inhibitors showed that both groups can be characterized as carboxylesterase, because bis-4-nitrophenyl phosphate inhibits the hydrolysis of both substrates, physostigmine has only a slight effect and EDTA is no inhibitor. The exception is the enzyme in the duodenum which is inhibited by all three inhibitors. The effect of phenobarbital induction and soman treatment on enzyme activity towards the two substrates were similar. Sex difference in the plasma activity towards methyl butyrate, but not 4-nitrophenyl butyrate, indicates that the group of carboxylesterase preferentially hydrolyzing 4-nitrophenyl butyrate may be the most important for the detoxification of soman.
Animals↗
The effect of methioninesulfoximine, an inhibitor of glutamine synthetase, on the levels of amino acids in the intact and decorticated rat neostriatum.
The glutamate terminals in rat rostral neostriatum were removed on one side by ipsilateral frontal cortical ablation. Intraperitoneal administration of L-methioninesulfoximine (MSO), an inhibitor of glutamine synthetase, induced different changes in amino acid concentration in the rostral neostriatum on the intact and decorticated sides. Glutamine was more reduced on the decorticated side than on the intact side. Glutamate changed less, but decreased more on the intact than on the lesioned side. There seems to be a correlation between the decrease in glutamine and the increase in alanine on both sides. GABA and taurine did not change significantly. The results indicate that the formation of glutamine probably formed from released glutamate is lower on the decorticated side than on the intact side.
Alanine↗
Substance P in subdivisions of the sciatic nerve, and in red and white skeletal muscles.
Substance P (SP) has been determined by radioimmunoassay in the sciatic nerve and its fibular, tibial and sural branches; and in a red, slow-twitch and a white, fast-twitch muscle supplied by the sciatic nerve. The mixed sciatic nerve contained 25 ng SP per g tissue wet wt., while the sural branch, which is supplying mainly skin, and thus is rich in sensory fibres, had a significantly higher content (49 ng/g). The mixed fibular and tibial branches contained approximately the same amount as the sciatic nerve proper. SP was also found in the skeletal muscles. The red m. soleus had a significantly higher content (0.61 ng/g) than the white m. extensor digitorum longus (0.22 ng/g). A dorsal root lesion which leads to degeneration of sensory fibres, reduced the SP level more than 90% both in nerves and muscles. Ventral root section, which leads to degeneration of somato-motor fibres and terminals, had, on the other hand, no effect. We conclude that sensory neurons are the only source of importance for SP in the sciatic nerve and in skeletal muscles. Based on the above findings, the possibility that SP may function as a mediator of an axon reflex in skeletal muscle is discussed.
Animals↗
Plasma renin activity and serum aldosterone during prolonged physical strain. The significance of sleep and energy deprivation.
Plasma renin activity (PRA), serum aldosterone and the serum and urinary levels of sodium and potassium have been investigated in 24 young men participating in a 5-day military training course with heavy continuous physical exercise, energy and sleep deprivation. The subjects were divided into three groups. Group 1 did not get any extra sleep or food, group 2 were compensated for the energy deficiency, and group 3 slept 3 h each night. The basic diet given to all the subjects was about 5,000 kJ and 2 g NaCl X 24 h-1 X cadet-1. The high calorie diet contained approximately 25,000-35,000 kJ and 20 g of NaCl X 24 h-1 X cadet-1. The study showed that serum aldosterone and PRA were extremely activated during such prolonged physical strain combined with lack of food and salt, whereas sleep deprivation did not seem to have any large influence. Only small variations were found in the serum levels of sodium and potassium and the urinary level of potassium during the course, whereas a decrease was seen in urinary sodium concentration. The fairly good correlations between the decrease in urinary sodium levels and the increase in PRA (r = 0.7) and further between PRA and serum aldosterone (r = 0.8) during the course indicate that there is a causal connection between the decrease in urinary sodium excretion and the increase in PRA and serum aldosterone.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗
Carboxylesterases, importance for detoxification of organophosphorus anticholinesterases and trichothecenes.
Several different types of experiments, including the use of inhibitors, have shown that carboxylesterases are a major factor in the metabolism and therefore detoxification of organophosphorus compounds such as soman and trichothecene toxins. The development of a new assay method for the enzyme has allowed us to separate the carboxylesterases into two major groups. The carboxylesterases can, however, be further separated by gel filtration, affinity chromatography, isoelectric focusing, and chromatofocusing into several isoenzymes. Liver microsomal carboxylesterases can be separated into five or six isoenzymes whereas guinea-pig plasma contains two isoenzymes. The isoenzymes differ in molecular weights, isoelectric points, substrate specificities, and affinity for inhibitors. Intravenous administration of a carboxylesterase preparation lowered the toxicity of soman in young rats. Carboxylesterases from rat and guinea-pig plasma inhibited by soman could be reactivated by DAM, whereas enzymes from porcine liver were not reactivated. Only one of the isoenzymes from rat liver microsomal preparation was responsible for the metabolism of T-2 toxin to HT-2. The further metabolism of HT-2 was performed by esterases from rat liver cytoplasma. Long-term exposure of the bronchial muscle to low concentration of soman modulate the bronchial contraction.
Animals↗
Esterase activities and soman toxicity in developing rat.
The activity of carboxylesterase and cholinesterase in plasma, liver and lung of young rats at different ages (5-31 days old) have been measured. The cholinesterase activity in the tissues of both sexes were almost constant during the development, and were similar to adult male activity. The carboxylesterase activity towards methyl butyrate and 4-nitrophenyl butyrate in plasma of both sexes increased from negligible (5 days old) to adult male value (31 days old). The carboxylesterase activities in liver increased markedly during the period, whereas the lung activities increased only slightly. The toxicity of soman was 6-7 fold higher in 5 days old rats compared to 30 days old rats. The decrease in toxicity correlated well with the increase in plasma carboxylesterase.
Aging↗
Changes in neurobiological parameters in the hippocampus after exposure to trimethyltin.
The effects of trimethyltin (TMT) on neurotransmitters, morphological changes and physiological activity of the hippocampus were studied. A single injection of TMT (8 mg/kg) decreased the high affinity uptake of glutamate (HA-Glu), which is a marker for glutamergic nerve terminals, after 7 days. The maximal reduction of HA-Glu was 42% and was obtained on postinjection day 21. Glutamate decarboxylase (GAD) and choline acetyltransferase (ChAT), markers for GABAergic and cholinergic structures, were not affected. The electrical activity of the hippocampus recorded through chronically implanted electrodes was altered by day three postinjection. The amplitude of the hippocampal electrographic record gradually decreased and the EEG ceased to be correlated with the rats' behavioral state. Fink-Heimer staining showed degenerating neurons within the subiculum, CA1, ventral CA3 and CA4 hippocampal subfields. TMT (3 mg/kg) injected once a week for three weeks decreased the HA-Glu significantly 21 days after the first injection. The HA-Glu was reduced by a maximum of 68%. The activity of ChAT was slightly increased only at day 35 postinjection while the GAD activity was not significantly reduced over a 21 day period beginning on day 14. Fink-Heimer staining showed degeneration of nerve cells within the CA1, ventral CA3 and CA4 hippocampal subfields. Both injection schedules produced degenerating neurons in the entorhinal cortex. The neurons of the dorsal CA3 region and the granule cells of the dentate gyrus were not lesioned by either TMT injection. The relationship between the behavioral, physiological and neurochemical changes induced by TMT will be discussed.
Animals↗
Effects of area 17 ablation on neurotransmitter parameters in efferents to area 18, the lateral geniculate body, pulvinar and superior colliculus in the cat.
The result of unilateral ablation of visual cortical area 17 in adult cats was consistent with glutamate-aspartate being the neurotransmitter in efferents to the lateral geniculate body, the pulvinar and the visual part of superior colliculus but not in efferents to area 18 and the non-visual strata of superior colliculus. Furthermore, the distribution of glutamatergic, GABAergic and cholinergic markers within the various subdivisions of the cat visual system complied well with observations made previously with biochemical, neurophysiological, histochemical and immunohistochemical methods, in this and other mammalian species.
Animals↗
Autoradiographic demonstration of glutamate structures after stereotaxic injection of kainic acid in rat hippocampus.
Autoradiography of high affinity uptake of L-glutamate in rat hippocampus after kainic acid lesion of CA3 and CA4 pyramidal cells has been used to identify the neurotransmitter of their ipsilateral connections. Glutamate or aspartate has tentatively been identified as the neurotransmitter of the projection from CA3 pyramidal cells to strata radiatum and oriens of CA1 and the projection from CA4 pyramidal cells to the inner parts of the molecular layer of area dentata.
Animals↗