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PubMed1989

Measurement of adrenolutin as an oxidation product of catecholamines in plasma.

Using the reverse phase high-performance liquid chromatography (HPLC) with mobile phases composed of simple acids, we have developed an assay technique for the measurement of adrenolutin, one of the oxidation products of catecholamines, in rat plasma. Ion-pairing chromatography permits the separation and quantitation of plasma adrenolutin (microM) in a linear manner. Sample preparation involved the precipitation of plasma proteins with perchloric acid and it is easier to handle a large number of samples at a time. However, we were unable to demonstrate the presence of adrenochrome, another oxidation product of catecholamines, in plasma since adrenochrome was rapidly destroyed in acid as well as in blood and was quickly changed into adrenolutin. Adrenolutin peak in HPLC was confirmed by 1) the retention time; 2) co-injection of adrenolutin and; 3) the appearance of 3H-adrenolutin after injection of 3H-norepinephrine. Administration of different catecholamines as well as adrenochrome and adrenolutin in rats also increased the level of adrenolutin in plasma. Adrenolutin was found to be present in plasma in other species including dog, rabbit and pig. High level of adrenolutin, which may represent total concentration of aminolutin in plasma, suggests the presence of an efficient mechanism for the oxidation of catecholamines under in vivo conditions.

Adrenochrome↗
PubMed1975

The effects of DL-p-chlorophenylalanine and DL-alpha-methyl-p-tyrosine on the brain catecholamine, serotonin and free amino acid contents in rat.

The changes in the brain levels of catecholamines, serotonin and free amino acids have been considered as a factor in the development of the physical dependence on and tolerance to some drugs. In order to show the relationships among the brain levels of these substances, three groups of rats were given i.p. DL-alphaMpT alone, DL-pCPA alone, or DL-alphaMpT and DL-pCPA together, twice a day for three days. Another group was kept as control. The brain levels of catecholamines, serotonin and free amino acids were determined. Although DL-alphaMpT and DL-pCPA alone caused a decrease in the levels of catecholamines and serotonin respectively, the administration of DL-alphaMpT and DL-pCPA together did not. The changes in the levels of free amino acids which were found were related to the metabolism of catecholamines and serotonin.

Amino Acids↗
PubMed1976

Effects of calcium and phosphate on catecholamines, ATP and dopamine beta-hydroxylase of chromaffin medullary granules.

Isolated bovine chromaffin granules lost their catecholamines to a significantly higher degree when incubated in isotonic sucrose-buffer of pH 7.4 containing 10 and 25 mM sodium phosphate, respectively, than during incubatation in sucrose with 4 mM sodium phosphate. 2. In 4mM sodium phosphate-sucrose, CaCl(2) in a final concentration of 1 and 2 mM, respectively, produced only traces of an amorphous precipitate of calcium phosphate which increased the efflux of catecholamines only to a small degree. The same concentrations of CaCl(2) in 10 and 25 mM sodium phosphate containing sucrose solutions caused larger amounts of calcium phosphate precipitate and simultaneously a very high efflux of catecholamines. Small amounts of exogenous ATP (1 mM) and MgCl(2) (0.3 mM) effectively antagonized the efflux of catecholamines from the granules evolked by calcium phosphate...

Adenosine Triphosphate↗
PubMed1980

Rise in blood pressure increases the release of endogenous catecholamines in the anterior hypothalamus of the cat.

In anaesthetized cats, anterior and posterior hypothalamic areas were simultaneously superfused with artificial CSF using two push-pull cannulae. The rates of release of endogenous catecholamines were determined in the superfusates which were continuously collected in periods of 10 seconds. In both areas, the rate of release of dopamine was higher than the rates of release of noradrenaline and adrenaline. Electrical stimulation of the splanchnic nerve elicited a rise in the arterial blood pressure and increased the rates of release of noradrenaline, adrenaline and dopamine in the anterior hypothalamic area. The rates of release of the catecholamines in the posterior hypothalamic area were not influenced by the pressor response to stimulation of the splanchnic nerve. Transection of the brain caudal to the hypothalamus reduced the rates of release of the catecholamines in the anterior hypothalamus and abolished the releasing effect of the stimulation of the splanchnic nerve. The results indicate that acute rise in blood pressure activates catecholamine cell bodies in the lower brain stem, which in turn stimulate the anterior hypothalamus to counteract the pressor response.

Animals↗
PubMed1981

Influence of acid-base alterations on myocardial sensitivity to catecholamines.

The influence of "respiratory" and "metabolic" acid-base alterations on the myocardial sensitivity to catecholamines was studied in the isolated rat atria. The ability of noradrenaline for increasing the atrial rate was enhanced during alkalosis and conversely, it was decreased by acidosis. These changes in sensitivity shifted the concentration-effect curve for noradrenaline to the right by about 0.5 log unit when the pH was lowered from 7.60 to 7.00. No changes in the maximum attainable response were detected. Essentially the same shifts of the concentration-effect curves were obtained with changes in pH brought about by altering the pCO2 or at constant pCO2. The decrease in the pH produced a similar shift to the right of the concentration-effect curve for isoprenaline, after the extraneuronal uptake inhibition by hydrocortisone and also in atria tissue with low content of endogenous noradrenaline (reserpine-pretreated and newborn rats). The ability of isoprenaline for increasing cyclic AMP levels in atrial tissue was also enhanced by alkalosis and decreased by acidosis. However, the shift to the right of the concentration-effect curve for cyclic AMP induced by the decrease in the pH was greater than the shift detected in the chronotropic-effect curve. In addition a decrease in the maximum increment of cyclic AMP was detected under acidosis, in spite of equal maximal chronotropic response. Our results support the hypothesis that the alterations in the sensitivity to catecholamines induced by the changes in pH are not due to a release of endogenous noradrenaline nor to alterations of the mechanisms which remove catecholamines from the biophase. The fact that cyclic AMP response to catecholamines was also reduced by acidosis strongly suggests that the mechanism(s) involved is located in the earlier steps of the events leading to the chronotropic effect of the beta-agonists.

Acid-Base Imbalance↗
PubMed1981

Liberation of catecholamines and 5-hydroxytryptamine from human blood-platelets.

Spontaneous and drug-induced liberation of 14C-5-hydroxytryptamine (14C-5HT), 3H-dopamine (3H-DA) and 3H-noradrenaline (3H-NA) from normal and reserpinized human blood-platelets has been determined from measurements of the amine contents before and after incubation in tris-buffer. In normal platelets the spontaneous liberation of 3H-catecholamines was more marked than that of 14C-5HT, but was less in percent for all these labelled amines than in reserpinized platelets. Thrombin lowered amine contents more in normal than in reserpinized platelets. The initial thrombin-induced decrease of 14C-5HT, in contrast to that of 3H-catecholamines, showed a partial recovery after 30 min which was abolished by imipramine. The benzoquinolizine Ro 4-1284 diminished all the amines in normal, but not in reserpinized platelets. In normal platelets tryamine affected 14C-5HT and 3H-DA about equally, whereas 3H-NA much less. Octopamine showed a similar pattern as tryamine, but was less potent. P-chlormethamphetamine (PCMA) and amphetamine decreased 3H-DA less markedly than 14C-5HT and 3H-NA not at all. In reserpinized platelets these arylalkylamines induced a decrease of 14C-5HT but not of 3H-catecholamines. It is concluded that (a) 3H-catecholamines like 14C-5HT are mainly localized in the granular pool of normal human platelets, (b) the pattern of action of a drug on intra- and extragranular amines depends not only on the nature of the drug and the amine to be liberated, but in comparison with previous results also on the species, (c) platelets are not completely satisfactory models for monoaminergic neurons, especially catecholaminergic ones regarding drug-induced amine liberation.

2H-Benzo(a)quinolizin-2-ol, 2-Ethyl-1,3,4,6,7,11b-↗
PubMed1986

Errors introduced by a tritium label in position 8 of catecholamines.

The neuronal and extraneuronal disposition of 3H-7,8- and 3H-7-labelled (-)-noradrenaline and dopamine was compared in in vitro studies. In agreement with earlier studies, the present results show that the presence of a tritium label in position 8 (i.e., on the alpha-carbon) has two consequences: a) the rate of deamination declines and b) part of the deamination results in the formation of an unlabelled aldehyde plus tritium water; tritium water is recovered from the OMDA-fraction of the column chromatographic procedure of Graefe et al. (1973). Whenever the deamination of a 3H-catecholamine is reduced (by tritium in position 8), the intraneuronal 3H-catecholamine concentration is increased. This increase, in turn, partly masks the decline in neuronal deamination (rat vas deferens). Irrespective of whether one determines the spontaneous efflux, the release of 3H-noradrenaline by nerve stimulation or the release of 3H-(-)-noradrenaline by the reserpine-like compound Ro 4-1284, the presence of tritium in position 8 distorts the results (experiments with rat vasa deferentia and/or rabbit aorta). In the extraneuronal system of the rat heart, two intracellular enzymes inactivate 3H-(-)-noradrenaline and 3H-dopamine: catechol-O-methyl transferase (COMT) and monoamine oxidase (MAO). Any hindrance of deamination (by tritium in position 8, COMT intact) leads to a shift of the metabolism of the 3H-catecholamines from the exclusively deaminated to the exclusively O-methylated metabolites. No differences between 3H-7,8- and 3H-7-labelled catecholamines were found after inhibition of MAO and COMT (extraneuronal accumulation and rate constant for efflux from the extraneuronal compartment III of the rat heart).(ABSTRACT TRUNCATED AT 250 WORDS)

2H-Benzo(a)quinolizin-2-ol, 2-Ethyl-1,3,4,6,7,11b-↗
PubMed1977

Smoking and catecholamine and c-AMP concentrations in the coronary circulation of man and the effect of oxprenolol.

Changes in catecholamine, c-AMP and lactate concentrations in the coronary circulation of man, during smoking, were studied in 12 patients. The heart rate increase from 63 +/- 2 beats/min (control) to 74 +/- 3 (smoking)(P less than 0.01), falling to 70 +/- 2 (10 min after smoking) (0.05 greater than P greater than 0.01), whilst coronary sinus c-AMP concentrations rose from 11 +/- 0.7 nmol/l (smoking) to 11.9 +/- 0.8 nmol/l (after smoking) (0.05 greater than P greater than 0.01; one tailed 't' test). There was no significant change in blood pressure, catecholamine or lactate concentrations. The study was repeated in eight of the patients following intravenous oxprenolol. Coronary sinus catecholamine concentrations increased from 4.1 +/- 0.7 nmol/l (control) to 5.5 +/- 1.1 nmol/l (after smoking) (0.05 greater than P greater than 0.01; one tailed 't' test), but heart rate and c-AMP concentrations remained unchanged, confirming that smoking-induced tachycardia is a result of a beta-adrenergic mechanism, at least part of which is due to a release of cardiac catecholamines. Arterial lactate concentrations increased only following oxprenolol from 0.74 +/- 0.07 mmol/l (control) to 0.83 +/- 0.09 mmol/l (smoking).

Adult↗
PubMed1990

Enhancement by cytochalasin B of ouabain-stimulated catecholamine secretion from cultured bovine adrenal chromaffin cells: possible relation to alteration in Na+/K(+)-pump activity.

1. Catecholamine secretion evoked by ouabain from cultured bovine adrenal chromaffin cells has previously been shown to be markedly enhanced by pretreatment of the cells with cytochalasin B (Morita et al., 1988). To elucidate a possible mechanism of this enhancement, the stimulatory action of ouabain on Ca2+ influx as well as catecholamine secretion was then examined in the cells pretreated with or without cytochalasin B. The effect of cytochalasin B pretreatment on the inhibitory action of ouabain on the Na+/K+ pump was also examined by measuring 86Rb+ uptake into the cells. 2. Pretreatment of the cells with cytochalasin B caused enhancement of ouabain-induced catecholamine secretion, and this enhancement was accompanied by the elevation of ouabain-stimulated 45Ca2+ uptake into the cells. The inhibitory action of ouabain on 86Rb+ uptake was significantly enhanced by pretreatment of the cells with cytochalasin B under the same conditions. 3. These results indicate that the enhancement of ouabain-induced catecholamine secretion caused by cytochalasin B pretreatment may be due to the increase in ouabain-stimulated Ca2+ influx into the cells and, furthermore, suggest the possibility that this increase in Ca2+ influx may be attributed to the potentiation of the inhibitory action of ouabain on the Na+/K+ pump in the adrenal chromaffin cell. Thus, the present study provides an evidence for a possible role of microfilaments as one of the intrinsic factors modulating the plasma membrane functions.

Adrenal Medulla↗
PubMed1994

Distribution of catecholamine(s) in the nervous system of a mermithid nematode, Romanomermis culicivorax.

The nervous system of the mermithid nematode Romanomermis culicivorax was investigated at different developmental stages for the distribution of catecholamine(s). The catecholaminergic nervous system increased in complexity during development. In the adult nematode, catecholamine(s) occurred in the nerve ring and associated ganglia, cephalic nerves, ventral and dorsal nerve cords and mid-body region ganglia. The distribution of catecholaminergic ganglia in the tail region was different for each sex. The tail region of males contained 16-20 catecholaminergic ganglia, whereas the tail of females contained only 2 ganglia. The catecholamine widely distributed throughout the nervous system of R. culicivorax is not dopamine, since immunoreactivity to dopamine was observed only in the amphids. The results suggest that the catecholamine(s) is widely distributed within the nervous system of R. culicivorax and might function as a neurohormone or neurotransmitter in controlling physiological and developmental processes.

Aedes↗
PubMed1991

Genotype differences in catecholamine concentrations in hypothalamus, intramedial hyperstriatum ventrale, and optic tectum of newly hatched chicks.

This study was designed to compare catecholamine concentrations among three brain areas of four pureline populations of visually isolated chicks. The purelines used were a commercial male line, a fertility selected line, an unselected fertility control line, and unselected White Jersey Giants. In general, male chicks had significantly larger brain weights than females. Six catecholamine-related compounds (norepinephrine, epinephrine, L-DOPA, dopamine, DOPAC, and MHPG) were measured via HPLC-ECD. No significant differences in neurochemical concentration were observed for any line or brain area due to sex of the chick. The hypothalamus (HT) contained the greatest concentration of catecholamines in all lines, followed by the intramedial hyperstriatum ventrale (IMHV) and optic tectum (OT). The HT exhibited consistent lateralization in all lines with the right HT containing ca. 30% more catecholamines than the left HT. While no consistent lateralization was observed among the other brain areas, the IMHV exhibited significantly different degrees of lateralization among the populations. Neuronal activity, as measured by MHPG:NE and DOPAC:DA ratio varied by line within each brain area. There were line differences for MHPG:NE in the HT, IMHV, and OT, while line differences for DOPAC:DA were observed in the HT. Since differences among purelines have been demonstrated in this study, care must be given to precisely define the genotype of chicks used in behavioral and neurochemical research.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1990

Correlation between carbohydrate and catecholamine level impairments in methionine sulfoximine epileptogenic rat brain.

This work shows that the convulsant methionine sulfoximine induces an increase in glucose and glycogen levels and a parallel decrease in norepinephrine and dopamine levels in rat brain. Among the epileptogenic agents, methionine sulfoximine is known to have a glycogenic property in the central nervous system. The aim of this work is to look for the neurochemical mechanism underlying this property. For this, catecholamines, glucose, and glycogen were measured at the same time in different areas of the brain in rats submitted to methionine sulfoximine. The convulsant induced an increase in glucose and glycogen levels as previously described and a decrease in dopamine and norepinephrine levels in all the areas of the rat brain. These changes were roughly dose dependent. When L-dihydroxyphenylalanine and benserazide (a decarboxylase inhibitor) were administered with methionine sulfoximine, the latter failed to induce seizures in rat up to 8 h after dosing. Moreover, the glucose and glycogen amounts did not increase. In all these experiments, there was an obvious evidence of parallelism between seizures, increase in carbohydrate levels, and decrease in catecholamine levels. These results allow to conclude that the glycogenic property of methionine sulfoximine in the central nervous system probably results from its ability to decrease norepinephrine and dopamine levels. Because the effect of the convulsant on the catecholamine levels persisted for long, it is normal that glucose and glycogen levels increased during preconvulsive, convulsive and postconvulsive period. Methionine sulfoximine is probably glycogenic in rat brain because it decreases catecholamine levels for a long time.

Animals↗
PubMed1993

Naphthalenesulfonamide derivatives ML9 and W7 inhibit catecholamine secretion in intact and permeabilized chromaffin cells.

The role of protein phosphorylation in catecholamine secretion from bovine adrenomedullary chromaffin cells was studied using different protein kinase inhibitors. Naphthalenesulfonamide derivatives as ML9 and ML7, more specific for the myosin light chain kinase, and the calmodulin antagonist W7 inhibited catecholamine secretion 20 and 40% respectively in digitonin-permeabilized chromaffin cells. ML9 also decreased calcium evoked protein phosphorylation of different proteins including tyrosine hydroxylase in permeabilized cells. These naphthalenesulfonamide derivatives showed also an effect in intact cells, ML9 and W7 produced 50% inhibition in catecholamine secretion and 45Ca2+ uptake, however H8 had no effect. The partial [3H]nitrendipine binding displacement of these drugs to adrenomedullary membranes suggests that these sulfonamide derivatives could interact directly with L-type calcium channels in intact cells. The results obtained in permeabilized cells suggest a possible role of protein phosphorylation in the regulation of catecholamine secretion in chromaffin cells.

Adrenal Glands↗
PubMed1985

An improved glyoxylic acid technique for the histochemical localization of catecholamines in brown adipose tissue.

A glyoxylic acid method using cryostat sections to demonstrate catecholaminergic fibres of the central nervous system was modified to show the extent of the adrenergic innervation in rat brown adipose tissue. It revealed prominent interlacing fluorescent parenchymal fibres surrounding individual adipocytes. The density of this network of fine fibres was not evident using earlier techniques. The new method also confirmed the dense networks of adrenergic fibres associated with arterial vessels. Its specificity was verified by simultaneously performing radioenzymatic determinations of tissue catecholamine levels and histochemical studies of brown adipose tissue from normal and sympathectomized rats. Chemical sympathectomy with 6-hydroxydopamine resulted in a pronounced decrease in brown adipose tissue and heart catecholamine (noradrenalin and dopamine) levels. Significantly, in brown adipose tissue of sympathectomized animals no fluorescence could be detected in terminal nerves of either the parenchyma or those of vascular smooth muscles. Nevertheless, some intense fluorescence was seen in axon bundles. The findings suggest that catecholamines of the parenchymal innervation form a larger proportion of the total catecholamine content of brown adipose tissue than was previously believed, provide stronger support for direct control of the function of multilocular adipocytes, and also confirm unpublished data reporting considerable dopamine content in brown adipose tissue.

Adipose Tissue, Brown↗
PubMed1983

Microfluorimetric quantification of catecholamine fluorescence in rat sympathetic ganglia.

The formaldehyde-induced fluorescence (FIF) technique was used to generate catecholamine fluorophores in the perikarya of the sympathetic neurons in the superior cervical ganglion of adult rats. During microfluorimetric quantification, the photodecomposition was eliminated by a rapid measuring procedure with a small excitation field and by using only visible light between the measurements. The catecholamine fluorescence, induced in protein microdroplets with increasing noradrenaline concentrations, was linear up to 2 X 10(-2) M which exceeds the noradrenaline content of even the most intensively fluorescent neurons. Thus, the differences in fluorescence intensities directly reflect the physiological state of each neuron with respect to their catecholamine content. The mean histograms reveal the changes which can only occur in certain neurons, and which can disappear if the mean only is assessed. The microfluorimetric method was sensitive enough to detect even minute changes induced by reserpine treatment in the catecholamine content of the sympathetic ganglion cells.

Animals↗
PubMed1990

Catecholamine-synthesizing enzymes and neuropeptides in rat heart epicardial ganglia; an immunohistochemical study.

The subepicardial atrial ganglia of rat hearts were examined using immunohistochemical techniques and antibodies against the catecholamine-synthetic enzymes tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH), and the neuropeptides substance P (SP), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP) and met-5-enkephalin (ENK). Some of the ganglion cells present in the ganglia exhibited DBH-like immunoreactivity (LI) and NPY-LI, whilst these cells never exhibited TH-, VIP-, CGRP-, SP- or ENK-LI. Groups of small cells exhibiting an intense TH-LI, corresponding to cells referred to as catecholamine-containing cells and sometimes small intensely fluorescent cells in the literature, were observed in the ganglia. A subpopulation of these cells exhibited immunoreactivity to one of the neuropeptides tested, namelyu SP. Only a few of the cells showing TH-LI displayed DBH-LI. Nerve fibres showing SP-, CGRP-, DBH- and TH-LI were present in the ganglia; some of these fibres being closely associated with the ganglion cells or with the cells showing TH-LI. The observation provide new information on the catecholamine-synthetic enzyme/neuropeptide expression of the ganglion and catecholamine-containing cells and of the associated nerve fibres of rat heart subepicardial ganglia.

Animals↗
PubMed1978

The paratrigeminal nucleus. II. Identification and inter-relations of catecholamine axons, indoleamine axons, and substance P immunoreactive cells in the neuropil.

The localization of three different putative neurotransmitters -- indoleamine, catecholamine, and Substance P -- was studied in the paratrigeminal nucleus of the rat and rhesus monkey at the light and electron microscope level by autoradiography following administration of [3H]5-hydroxytryptamine, or [3H]norepinephrine, and by immunocytochemistry using the unlabelled anti-Substance P antiserum peroxidase--antiperoxidase technique. The paratrigeminal neurons are not monoaminergic but certain cells exhibit Substance P-like immunoreactivity. These cells receive a rich plexus of indoleamine afferents, a sparse catecholamine input, and a rich plexus of fibres with Substance P-like immunoreactivity. Of the entire monoaminergic population of labelled axons, more than 60% are synaptic and less then 40% nonsynaptic, and this proportion is the same for indoleamines as for catecholamines. Indoleamine axons form a heterogeneous population with at least four different morphological types that are synaptic and three that are nonsynaptic. They bear distinctive collections of small, clear, tubular or large granular vesicles, which distinguish one category of axon from another. These axons engage in numerous axo--somatic, axo--spinous, axo--dendritic, and possibly axo--axonic relations with paratrigeminal neurons. The catecholamine axons are also heterogeneous in axoplasmic morphology but their terminal contacts are distributed to more peripheral portions of dendrites. The significance of the inter-relations between the monaminergic and peptidergic elements in the paratrigeminal nucleus is discussed in relation to the possible functions of this nucleus as a nociceptive, chemosensitive, or pressure-sensitive centre on the lateral medullary surface.

Animals↗
PubMed1989

Localization of phospholipids in human catecholamine neurons with Baker's acid haematein.

Protein bodies in catecholamine neurons of the normal human brain contain arginine-rich proteins similar to those present in the core of Lewy bodies in Parkinsonian brains. Lewy bodies are known also to contain phospholipids in their core, demonstrable with Baker's acid haematein. We used Baker's procedure on catecholamine neurons of normal brains to test whether the protein bodies also contain phospholipids. Postmortem tissues of three control individuals were used in this study. Locus coeruleus and substantia nigra were dissected out from the fresh brains and two sets of blocks were made from each area. One set was fixed in formol-calcium for the preservation of lipids, the second was fixed in Bouin's solution and treated with hot pyridine for the extraction of lipids. Finally, both sets were subjected to the same chroming and staining procedure according to Baker. In catecholamine neurons the protein bodies were stained with acid haematein, indicating the presence of phospholipids. Since this staining resisted pyridine extraction we conclude that these phospholipids are firmly bound to the basic proteins. Thus, protein bodies in healthy catecholamine neurons give the same positive reaction for phospholipids as that reported for the core of Lewy bodies in damaged neurons in Parkinsonism.

Aged↗