Neuropeptide tyrosine (NPY)-induced potentiation of the pressor activity of catecholamines in conscious rats

IV bolus administration of 2.5-50 micrograms NPY (0.6-12.5 nmol) to conscious rats produced a dose- and time-dependent increase in systolic and diastolic blood pressure. Following priming with 2.5 micrograms NPY, or larger doses, the subsequent administrations of noradrenaline produced pressor responses that were potentiated both in magnitude and duration. The NPY-induced potentiation of the pressor response to noradrenaline was dose-dependent and extended to the pressor action of adrenaline and angiotensin II but not to the hypotensions produced by bradykinin or isoproterenol. The potentiation was not related to the fact that multiple doses of catecholamines were repeated. Reserpine did not substantially modify the NPY-induced potentiation of the pressor activity of the catecholamines. Chemical sympathectomy following 6-hydroxydopamine caused a marked supersensitivity to the catecholamines and NPY but obliterated the NPY-induced potentiation of the pressor effect of adrenaline. Nifedipine reduced the pressor action of the catecholamines and NPY but did not attenuate the NPY-induced potentiation of the pressor action of catecholamines. It is concluded that the acute pressor effect of NPY and of the potentiation of the catecholamine pressor effects involve different mechanisms.     

Hormonal regulation of fat esterification & oxidation in hepatocytes from fed rats: modulation by 3-mercaptopicolinate

3-Mercaptopicolinate, an inhibitor of phosphoenolpyruvate carboxykinase (PEPCK), decreased esterification of [1-14C] oleate and [1-14C] myristate in hepatocytes from fed rats. In the absence of 3-mercaptopicolinate, adrenaline, noradrenaline, vasopressin or angiotensin II increased esterification to triacylglycerol of [1-14C] oleate but not [1-14C] myristate. Cyclic AMP decreased esterification of both oleate and myristate. In the presence of 3-mercaptopicolinate, stimulation of oleate esterification by the catecholamines, vasopressin or angiotensin II was increased, and stimulatory effects of these hormones on myristate esterification were observed. Adrenaline, noradrenaline, vasopressin or angiotensin II increased 14CO2 production from both [1-14C] oleate and [1-14C] myristate but the degree of stimulation was similar in the absence or presence of 3-mercaptopicolinate. The results indicate a role for the catecholamines and angiotensin II in the regulation of liver fat metabolism and emphasize the potential importance of changes in activity of PEPCK as determinants of hepatic carbon flux.     

Effects of temperature and oxygen availability on circulating catecholamines in the toad Bufo marinus.

The release of catecholamines during hypoxia has received limited attention in amphibians and the adrenergic regulation of cardio-pulmonary functions is, therefore, not well understood at the organismic level. To describe the changes in plasma catecholamine concentrations, we exposed toads (Bufo marinus) to different levels of hypoxia at two temperatures (15 and 25 degrees C). In addition, blood oxygen binding properties were determined in vitro at 15 and 25 degrees C at two different pH values. Hypoxia elicited a significant increase in plasma catecholamines (adrenaline and noradrenaline) at both temperatures, in spite of a respiratory alkalosis. At 15 degrees C, the increase was from 2.6+/-1.0 in normoxia to 4.8+/-1.4 ng ml(-1) at an inspired oxygen fraction of 0.05. At 25 degrees C, the hypoxic release of catecholamines was significantly higher (maximum levels of 44.8+/-11.6 ng ml(-1)). Plasma noradrenaline concentration was elevated at the most severe hypoxic levels, suggestive of an adrenal release. The arterial oxygen threshold for catecholamine release were approximately 1.0 mmol O(2) l(-1) blood or a PaO(2) of 30 mmHg. The P(50) values at 15 degrees C were 23.5+/-0.7 and 28.9+/-1.0 mmHg at pH 7.98+/-0.01 and 7.62+/-0.02, respectively, and increased to 36.5+/-0.6 and 43.0+/-1.1 mmHg at pH 8.04+/-0.04 and 7.67+/-0.05, respectively, at 25 degrees C. The oxygen equilibrium curves were linear when transformed to Hill-plots and Hills n (the haemoglobin subunit co-operativity) ranged between 2.24 and 2.75. The in vitro blood O(2) binding properties corresponded well with in vivo data.     

Catecholamine sulfates: end products or metabolic intermediates?

Dopamine-beta-hydroxylase catalyzes the beta-oxidation of dopamine to noradrenaline while phenylethanolamine-N-methyltransferase converts noradrenaline to adrenaline. Since catecholamine sulfates represent the predominant form of catecholamines in human tissues, we have studied the role of dopamine sulfate and noradrenaline sulfate as alternate substrates for dopamine-beta-hydroxylase and phenylethanolamine-N-methyltransferase, respectively. Dopamine 3-sulfate, dopamine 4-sulfate and noradrenaline 3-sulfate were chemically synthesized and exhaustively purified by ion-exchange chromatography. Dopamine-beta-hydroxylase and phenylethanolamine-N-methyltransferase were partially purified from human adrenals. Using tyramine as substrate, dopamine-beta-hydroxylase is slightly inhibited by dopamine 3-sulfate according to some irreversible or mixed mechanisms. When dopamine-beta-hydroxylase was incubated with dopamine 3-sulfate or dopamine 4-sulfate, we were not able to find any synthesis of either noradrenaline sulfate or free noradrenaline. Using phenylethanolamine as substrate, the enzymatic activity of phenylethanolamine-N-methyltransferase remains unchanged with addition of dopamine 3-sulfate, dopamine 4-sulfate or noradrenaline 3-sulfate. It was concluded that dopamine sulfate is not an alternate substrate for either dopamine-beta-hydroxylase or phenylethanolamine-N-methyltransferase nor is noradrenaline 3-sulfate an alternate substrate for phenylethanolamine-N-methyltransferase.     

The effects of repeated physical stress or fasting on catccholamine storage and release in the rainbow trout, Oncorhynchus mykiss

Rainbow trout, Oncorhynchus mykiss, were subjected to either physical stress (twice daily chasing to exhaustion for 5 days) or a period of 2 months of fasting. Following these treatments, the levels of catecholamines, adrenaline and noradrenaline, stored within the kidney and posterior cardinal vein (PCV) were determined. The ability of the catecholamine-storing chromaffin cells to release catecholamines in response to cholinergic stimulation was measured using an in situ saline-perfused PCV preparation. In the physically stressed fish, the concentration (μg catecholamine g^?1 tissue) of noradrenaline within the anterior and middle thirds of the kidney increased; the concentration of adrenaline was unchanged in all tissues. The content (μg) of noradrenaline or adrenaline, within the various tissues, was similar in both groups of fish with the exception of a higher noradrenaline content in the middle third of the kidney in the physically stressed fish. The total catecholamine content (μg catecholamine) of these tissues (kidney+PCV) was unaffected by physical stress. With the exception of a lower concentration of adrenaline in the middle third of the kidney, the concentrations of catecholamines were unaffected by fasting. There was a trend towards a greater content (μg) of noradrenaline within all of the tissue regions of the fed fish, however, a significant difference was only observed in the anterior third of the kidney. The content of adrenaline in the fed fish was greater in all regions of the kidney as well as the middle third of the PCV. The total catecholamine content (kidney + PCV) was lower in the fasted fish owing to significantly lower PCV and kidney masses. Prolonged physical stress caused a decrease in the responsiveness of the chromaffin cells to the cholinoceptor agonist carbachol (10^?8 to 10^?4mol). The ED₅₀ (the dose of carbachol required to elicit a half maximal response) for catecholamine release was 0·96 ± 10^?6mol carbachol in the physically stressed fish and 0·84 ± 10^?7 in the control fish. Fasting did not alter the pattern of catecholamine release. The ED₅₀ values were 0·96 ± 10^?7 and 1·24 ± 10^?7 mol for fasted and fed fish, respectively. Thus, a physical stress affected both catecholamine storage and release whereas fasting affected only storage and not the release process.     

Effects of catecholamines on rat myocardial metabolism. II. Influence of catecholamines on 32p-incorporation into rat myocardial adenylic nucleotides and their turn-over.

1. The influence of catecholamines (adrenaline and noradrenaline) on 32Pi incorporation into intracellular phosphate and adenylic nucleotides has been studied on rat myocardium slices; consequently, the turn-over of nucleotides could be determined and compared under the influence of these two hormones. 2. In order to specify the site of action of these catecholamines, several inhibitors and activators of energetic metabolism were included in the incubation medium: 3'5'-AMP, caffein, ouabain, oligomycin, rotenone + antimycin. 3. Both catecholamines favour Pi exchanges between intra and extracellular spaces; ATP turn-over is greatly increased, while ADP turn-over is slightly decreased, and 32P-incorporation into ADP is increased. 4. 3'5'-AMP and caffein are without effect on Pi penetration; however, caffein increases catecholamine effects on this penetration. ATP turn-over is slightly increased by 3'5'-AMP or caffein. 5. Ouabain decreases ATP turn-over but does not prevent the adrenaline induced acceleration. Inhibitors of oxidative phosphorylation and electron transport decrease ATP-turn-over severely; this inhibition is not released by catecholamines. 6. It is concluded that the catecholamine effects observed are dependent on the oxidative phosphorylations process. The increase of Pi exchange by catecholamines may be related to the increase of extracellular space and cation translocations we observed with the hormones.     

Sodium Fluoride Mimics the Effect of Prostaglandin E2 on Catecholamine Release from Bovine Adrenal Chromaffin Cells

We have reported recently that prostaglandin E2 (PGE2) stimulated phosphoinositide metabolism in bovine adrenal chromaffin cells and that PGE2 and ouabain, an inhibitor of Na+, K(+)-ATPase, synergistically induced a gradual secretion of catecholamines from the cells. Here we examined the involvement of a GTP-binding protein(s) in PGE receptor-induced responses by using NaF. In the presence of Ca2+ in the medium, NaF stimulated the formation of all three inositol phosphates, i.e., inositol monophosphate, bisphosphate, and trisphosphate, linearly over 30 min in a dose-dependent manner (15-30 mM). This effect on phosphoinositide metabolism was accompanied by an increase in cytosolic free Ca2+. NaF also induced catecholamine release from chromaffin cells, and the dependency of stimulation of the release on NaF concentration was well correlated with those of NaF-enhanced inositol phosphate formation and increase in cytosolic free Ca2+. Although the effect of NaF on PGE2-induced catecholamine release in the presence of ouabain was additive at concentrations below 20 mM, there was no additive effect at 25 mM NaF. Furthermore, the time course of catecholamine release stimulated by 20 mM NaF in the presence of ouabain was quite similar to that by 1 microM PGE2, and both stimulations were markedly inhibited by amiloride, with half-maximal inhibition at 10 microM. Pretreatment of the cells with pertussis toxin did not prevent, but rather enhanced, PGE2-induced catecholamine release over the range of concentrations examined. These results demonstrate that NaF mimics the effect of PGE2 on catecholamine release from chromaffin cells and suggest that PGE2-evoked catecholamine release may be mediated by the stimulation of phosphoinositide metabolism through a putative GTP-binding protein insensitive to pertussis toxin.     

Differences in regional brain catecholamine metabolism after a decrease in blood pressure

Using a sensitive radiometric assay for determination of catecholamines, the α-methyl-para-tyrosine-induced catecholamine disappearance was studied in discrete brain regions of rats after acute hypotension. The decrease in blood pressure was induced either by controlled haemorrhage or by the administration of guanethidine. In both cases an increase of the α-methyl-para-tyrosine-induced noradrenaline disappearance was observed in the nucleus commissuralis and the A₁-region in the medulla oblongata. In the paraventricular nucleus the disappearance of noradrenaline as well as that of adrenaline was enhanced. The results are in support for the involvement of catecholamines in particular brain regions in the regulation of blood pressure.     

Oxidative stress in the neurodegenerative diseases--potential antioxidant activity of catecholamines

Neurodegenerative diseases (e.g., Alzheimer's Disease, Parkinson's Disease and Amyotrophic Lateral Sclerosis), are characterized by the deposition of misfolded proteins (alpha beta-peptide, alpha-synucleine and Cu, Zn superoxide dismutase, respectively). Oxidative stress is accompanied by the diminished level of catecholamines in neuronal tissue. Beside their role in transduction of nervous stimuli, catecholamines might act as antioxidants. Herein, we focused on the antioxidant activity of catecholamine neurotransmitters--dopamine (and its precursor L-DOPA), adrenaline and noradrenaline. There is an increasing evidence that catecholamines might act as scavengers of free radicals and experimental data indicate the antioxidant potency of catecholamines. However, the exact mechanism of this action is not defined.     

Combined Stereological and Biochemical Analysis of Storage and Release of Catecholamines in the Adrenal Medulla of the Rat

Adrenal medullae from rats injected with insulin 1 h prior to decapitation were analyzed by stereology and their catecholamine content was determined. In control animals the ratio between adrenaline- and noradrenaline- containing cells was 4.06 whereas the ratio between adrenaline and noradrenaline contents was 4.11. The glands from insulin-treated animals showed a 45% reduction in adrenaline content and a 42% decrease in the volumetric density of the adrenaline-containing granules. However, neither the noradrenaline content nor the volumetric density of noradrenaline-containing granules was modified in glands from insulin-treated animals. From these data, the amount and concentration of adrenaline or noradrenaline in a single granule have been calculated. The results are consistent with the view that chromaffin granules are the source of the released catecholamines. The present paper demonstrates that stereology combined with biochemistry is a useful tool for resolution of problems at the cellular and subcellular levels.     

Neuromediator mechanisms of the effect of the antihistamine agent dimebone on the brain

Dimebone was shown to inhibit monoamine oxidase (MAO) deaminating dopamine and serotonin, decrease dopamine metabolism in the basal ganglia of the rat brain, increase noradrenaline level and depress dopamine deamination in the hypothalamus. Dimebone first increased and then diminished the release of dopamine in the cortex, with the concomitant MAO activation and the increase in dopamine and noradrenaline levels. The in vitro experiments have demonstrated that dimebone (10(-4)) preferentially inhibited MAO activity, type B and dopamine deamination in homogenates of different rat brain structures. The role of MAO inhibition in the mechanism of dimebone action on the catecholamine metabolism in the brain structures and its stimulating effect on CNS are discussed.     

The actions of vasoactive compounds in the foetus and the effect of perfusion through the placenta on their biological activity

In acute experiments on the in utero foetal lamb, angiotensin II was a more potent pressor agent that either noradrenaline or adrenaline, and the response to angiotensin II was not consistently modified by the combined administration of alpha and beta-adrenergic blocking agents. A significant reduction in the pressor response of the foetus to angiotensin II and noradrenaline occurred with infusion of these compounds to the foetus by the umbilical artery when compared with the response obtained with infusions of the same doses of these drugs by the umbilical vein. Moreover, the concentration of angiotensin II (pg. ml-1) present in the foetal circulation was less following umbilical arterial infusions compared with umbilical vein infusions of the same doses. A similar reduction in the pressor activity of adrenaline and the cardio-stimulant effect of isoprenaline occurred when these drugs were infused by the umbilical artery. It is concluded that the foetus, like the adult animal, is more sensitive to angiotensin II than to catecholamines and that the biological activities of noradrenaline, angiotensin II, adrenaline and isoprenaline are reduced by perfusion through the foetal placenta.     

Regulation of atrial contraction in the seawater-adapted eel,Anguilla japonica

The atrium isolated from the seawater-adapted eel beats spontaneously in normal Ringer solution for more than 10 hr. The strength of beating was inhibited by acetylcholine (ACh) and the inhibitory effects were blocked by atropine, a muscarinic ACh-receptor antagonist, indicating existence of muscarinic ACh-receptor on the atrium. The atrial contractility was stimulated by catecholamines and their agonists; the order of potency being isoproterenol > adrenaline (AD) = noradrenaline (NA) > phenylephrine > clonidine. The stimulatory effects of AD was completely blocked by propranolol, a β-adrenoceptor antagonist, but not by phentolamine, an α-adrenoceptor antagonist. These data were consistent with characteristics of β-adrenoceptors. Further characterization of the β-receptor was not attempted. The positive inotropic and chronotropic actions of AD were not completely blocked either by atenolol, a β₁-adrenoceptor antagonist, or by ICI 118551, a β₂-adrenoceptor antagonist. When electrical current with a short duration (0.25 msec) was passed through the atrium, the beating was inhibited initially, then enhanced later. The initial inhibition was inhibited by atropine and the later enhancement was blocked by propranolol. These results indicate that the electrical stimulation releases ACh and catecholamine(s) from the nerve endings. The positive inotropic and chronotropic effects of catecholamines were mimicked by tyramine, a catecholamine releaser from sympathetic nerve endings.     

Effect of a fall of blood pressure on the release of catecholamines in the hypothalamus

The posterior hypothalamus of anaesthetized cats was superfused through a push-pull cannula and the release of endogenous catecholamines was determined in the superfusate which was continuously collected in 15 min periods. Fall in blood pressure elicited by nitroprusside or bleeding led to an increased rate of release of noradrenaline, adrenaline and dopamine in the hypothalamus. Transection of the brain causal to hypothalamus greatly reduced the rate of resting release of the catecholamines and abolished the enhancing effects of bleeding and nitroprusside. Determination of the catecholamines in samples which were collected in 90 s periods suggested a different pattern of release of the three catecholamines. Further shortening of the collection period (10 s) showed that the fall in blood pressure immediately increased the release of dopamine, while the rates of release of noradrenaline and adrenaline were increased gradually. Hypotension did not influence the rates of release of the catecholamines in the anterior hypothalamus. It is concluded that dopamine, adrenaline and noradrenaline systems of the hypothalamus are involved in the regulation of the arterial blood pressure. The different patterns of release might indicate that dopamine exerts a different function from those of noradrenaline and adrenaline in the normalization of the blood pressure after acute hypotension.     

Influence of adrenoceptors on thrombin-induced phosphoinositide metabolism in rat platelets

Stimulation of washed rat platelets with thrombin resulted in an increased turnover of phosphoinositides. Adrenaline and isoproterenol both inhibited thrombin-induced phosphatidic acid formation in a dose-dependent manner. Inhibitory responses of both compounds were blocked by a beta-adrenoceptor antagonist. However, isoproterenol was a more potent inhibitor than adrenaline. Addition of a selective alpha2-adrenoceptor antagonist potentiated the inhibitory effect of adrenaline up to the level observed with isoproterenol. Prestimulation of beta-adrenoceptors with isoproterenol, followed by addition of adrenaline (or noradrenaline) markedly diminished the inhibitory effect induced by the full beta-adrenoceptor agonist. Our results indicate that, in rat platelets, catecholamines are able to counteract, via alpha2-receptors, the beta-adrenoceptor-mediated inhibition of thrombin-induced phosphatidic acid formation. This suggests that catecholamines, by controlling cAMP level, may modulate phospholipase C activity and thereby platelet reactivity.     

Effects of Opioid Peptides Containing the Sequence of Met5-Enkephalin or Leu5-Enkephalin on Nicotine-Induced Secretion from Bovine Adrenal Chromaffin Cells

Eighteen endogenous opioid peptides, all containing the sequence of either Met5- or Leu5-enkephalin, were tested for their ability to modify nicotine-induced secretion from bovine adrenal chromaffin cells. ATP released from suspensions of freshly isolated cells was measured with the luciferin-luciferase bioluminescence method as an index of secretion. None of the peptides affected 5 microM nicotine-induced ATP release at 10 nM. Three peptides inhibited secretion at 5 microM: dynorphin1-13, dynorphin1-9, and rimorphin inhibited by 65%, 37%, and 29% respectively. Use of peptidase inhibitors (bestatin, thiorphan, bacitracin, or 1,10-phenanthroline) did not result in any of the other peptides showing potent actions on the nicotinic response, although bestatin and thiorphan did enhance the inhibitory actions of dynorphin1-13 and dynorphin1-9 by 20-30%. Nicotine-induced secretion of endogenous catecholamines from bovine chromaffin cells cultured for 3 days was also studied to assess any selective actions of the peptides on adrenaline or noradrenaline cell types. Dynorphin1-13 was 1,000-fold more potent than Leu5-enkephalin at inhibiting endogenous catecholamine secretion. Dynorphin1-13 was slightly more potent at inhibiting noradrenaline release than adrenaline release whereas Leu5-enkephalin showed the opposite selectivity. The structure-activity relationships of opioid peptide actions on the chromaffin cell nicotinic response are discussed in relation to the properties of the adrenal opioid binding sites.     

Catecholamine levels in whole brain of stressed and control domestic and wild rats (Rattus Norvegicus)

Whole brain catecholamine (dopamine, nonadrenaline, adrenaline) levels were measured in control and electric footshocked Wild and domestic (Sprague Dawley and Long Evans) rats. No significant differences were found among the three strains of rats for combined total catecholamine content, or for combined total catecholamines between the control and footshocked groups. Significant differences were found for the total of each catecholamine taken separately, dopamine being present at three times the level of noradrenaline and ten times the level of adrenaline. No significant differences were found for dopamine in both control and footshocked animals among all three rat strains. Noradrenaline was significantly higher in the control domestic rats compared to the control Wild rats, and in the footshocked domestic rats compared to the footshocked Wild rats. No differences in noradrenaline levels were found between Sprague Dawley and Long Evans rats, but noradrenaline increased significantly in the latter following footshock. Adrenaline was significantly highest in the Sprague Dawley controls and lowest in the Wild controls. Footshocking resulted in almost identical levels of adrenaline in the domestic strains and an increase in the F₁ Wild strain.     

Metabolic responses to catecholamines in dogs injected with a single dose of triiodothyronine.

Lipolytic and glycogenolytic responses to catecholamine infusions were studied in resting dogs before and 20 h following administration of a single dose (0.1 mg/kg) of triiodothyronine (T3). In the dogs pretreated with T3 much higher increases in the plasma FFA concentration were found both during noradrenaline and adrenaline infusions in comparison with control experiments. Adrenaline-induced increases in blood LA and glucose levels were also significantly higher in T3-pretreated dogs than in controls. The blockade of beta-adrenergic receptors with propranolol prevented the increases in blood FFA and LA concentrations during subsequent adrenaline infusion. Phentolamine -- the alpha-adrenergic blocking agent -- infused to the T3-pretreated dog inhibited the adrenaline-induced rise in blood glucose level. The observed changes in the metabolic responses to catecholamines induced by triiodothyronine pretreatment indicate that at least in the dog this hormone potentiates both the lipolytic and glycogenolytic effects of catecholamines acting on appropriate adrenergic receptors.     

Enzymatic Mechanisms Involved in Phenanthrene Degradation by the White Rot Fungus Pleurotus ostreatus

The enzymatic mechanisms involved in the degradation of phenanthrene by the white rot fungus Pleurotus ostreatus were examined. Phase I metabolism (cytochrome P-450 monooxygenase and epoxide hydrolase) and phase II conjugation (glutathione S-transferase, aryl sulfotransferase, UDP-glucuronosyltransferase, and UDP-glucosyltransferase) enzyme activities were determined for mycelial extracts of P. ostreatus. Cytochrome P-450 was detected in both cytosolic and microsomal fractions at 0.16 and 0.38 nmol min(sup-1) mg of protein(sup1), respectively. Both fractions oxidized [9,10-(sup14)C]phenanthrene to phenanthrene trans-9,10-dihydrodiol. The cytochrome P-450 inhibitors 1-aminobenzotriazole (0.1 mM), SKF-525A (proadifen, 0.1 mM), and carbon monoxide inhibited the cytosolic and microsomal P-450s differently. Cytosolic and microsomal epoxide hydrolase activities, with phenanthrene 9,10-oxide as the substrate, were similar, with specific activities of 0.50 and 0.41 nmol min(sup-1) mg of protein(sup-1), respectively. The epoxide hydrolase inhibitor cyclohexene oxide (5 mM) significantly inhibited the formation of phenanthrene trans-9,10-dihydrodiol in both fractions. The phase II enzyme 1-chloro-2,4-dinitrobenzene glutathione S-transferase was detected in the cytosolic fraction (4.16 nmol min(sup-1) mg of protein(sup-1)), whereas aryl adenosine-3(prm1)-phosphate-5(prm1)-phosphosulfate sulfotransferase (aryl PAPS sulfotransferase) UDP-glucuronosyltransferase, and UDP-glucosyltransferase had microsomal activities of 2.14, 4.25, and 4.21 nmol min(sup-1) mg of protein(sup-1), respectively, with low activity in the cytosolic fraction. However, when P. ostreatus culture broth incubated with phenanthrene was screened for phase II metabolites, no sulfate, glutathione, glucoside, or glucuronide conjugates of phenanthrene metabolites were detected. These experiments indicate the involvement of cytochrome P-450 monooxygenase and epoxide hydrolase in the initial phase I oxidation of phenanthrene to form phenanthrene trans-9,10-dihydrodiol. Laccase and manganese-independent peroxidase were not involved in the initial oxidation of phenanthrene. Although P. ostreatus had phase II xenobiotic metabolizing enzymes, conjugation reactions were not important for the elimination of hydroxylated phenanthrene.     

Endogenous Noradrenaline and Dopamine in Nerve Terminals of the Hippocampus: Differences in Levels and Release Kinetics

The presence and release of endogenous catecholamines in rat and guinea pig hippocampal nerve terminals was studied by fluorimetric HPLC analysis. In isolated nerve terminals (synaptosomes) the levels and breakdown of endogenous catecholamines were determined and the release process was characterized with respect to its kinetics and Ca2+ and ATP dependence. Endogenous noradrenaline and dopamine, but not adrenaline, were detected in isolated hippocampal nerve terminals. For dopamine both the levels and the amounts released were more than 100-fold lower than those for noradrenaline. In suspension, released endogenous catecholamines were rapidly broken down. This could effectively be blocked by monoamine oxidase inhibitors, Ca(2+)-free conditions, and glutathione. The release of both noradrenaline and dopamine was highly Ca2+ and ATP dependent. Marked differences were observed in the kinetics of release between the two catecholamines. Noradrenaline showed an initial burst of release within 10 s after K+ depolarization. The release of noradrenaline was terminated after approximately 3 min of K+ depolarization. In contrast, dopamine release was more gradual, without an initial burst and without clear termination of release within 5 min. It is concluded that both catecholamines are present in nerve terminals in the rat hippocampus and that their release from (isolated) nerve terminals is exocytotic. The characteristics of noradrenaline release show several similarities with those of other classical transmitters, whereas dopamine release characteristics resemble those of neuropeptide release in the hippocampus but not those of dopamine release in other brain areas. It is hypothesized that in the hippocampus dopamine is released from large, dense-cored vesicles, probably colocalized with neuropeptides.