Evidence for participation of catecholamines in cardiac action of ouabain: Positive chronotropic effect

1 The shortening of cycle length (=positive chronotropic effect) by ouabain produced in isolated spontaneously beating atria of the guinea-pig was analyzed.2 The action of ouabain was dose-dependent; threshold response was seen at 1 x 10(-7) M, and maximal response occurred at 4 x 10(-7) M. The half-time of the ouabain effect was about 20 minutes.3 The positive chronotropic effect of ouabain was reduced to 40% by beta-adrenoceptor blockade (3.3 x 10(-9) M propranolol) or by reserpine-depletion of catecholamines. Incubation of reserpine-treated atria with noradrenaline partially restored the action of ouabain.4 The effect of ouabain was greatly dependent upon the calcium concentration. The optimal calcium level was 2.5 x 10(-3) M. Calcium and ouabain acted synergistically.5 Increasing calcium concentrations inhibited the positive chronotropic effect of noradrenaline in a manner similar to increasing ouabain concentrations.6 A hypothesis is proposed which explains the chronotropic effect of ouabain on the basis of two mechanisms: (1) increase of the catecholamine concentration affecting the pacemaker; (2) mobilization of calcium, i.e. increase of the biologically effective intracellular calcium level.     

The effects of drugs on pacemaker regions of isolated rabbit renal pelvis

Acetylcholine failed to change the frequency of spontaneous contractions in the proximal region of isolated rabbit renal pelvis, but significantly increased the contractile frequency in the middle and distal regions, which then reached similar levels to those of the proximal region. Pretreatment with reserpine caused a decrease in the frequency of spontaneous contractions in the proximal, but not in the middle or distal regions. Reserpine-pretreated tissues developed hypersensitivity to catecholamines, while acetylcholine produced effects similar to those observed in control preparations. Atropine and N-methyl-scopolamine antagonized the action of acetylcholine in both the middle and the distal regions, suggesting that the action was exerted through muscarinic receptors. Adrenaline and alpha-stimulating drugs, but not isoprenaline, significantly increased the contractile frequency of all three tissues: the increase in the proximal region reached levels in excess of its fundamental maximal frequency. Phentolamine caused a significant decrease in the frequency of the proximal region and fully inhibited the stimulating action of catecholamines, indicating that this stimulation was mediated by alpha-adrenoceptors. The myogenic nature of pacemaking cells was confirmed by the effect of tetrodotoxin, ouabain and verapamil. The decrease in frequency in the pacemaker region of the proximal pelvis whether caused by reserpine or phentolamine indicates a significant role of catecholamines in modulating pacemaker activity.     

Modulation by ouabain and diphenylhydantoin of veratridine-induced 22Na influx and its relation to 45Ca influx and the secretion of catecholamines in cultured bovine adrenal medullary cells

Summary The effects of ouabain and diphenylhydantoin on the secretion of catecholamines induced by veratridine were investigated in cultured bovine adrenal medullary cells with special reference to ion fluxes. Veratridine itself induced an influx of ²²Na and ⁴⁵Ca as well as secretion of catecholamines, which were antagonized by tetrodotoxin, a selective inhibitor of voltage dependent Na channels. The secretion of catecholamines caused by veratridine was not observed either in Na free or Ca free medium. Veratridineinduced influx of ⁴⁵Ca did not occur in Na free medium, while veratridine-induced influx of ²²Na occurred even in Ca free medium. Veratridine-induced influx of ²²Na, ⁴⁵Ca and secretion of catecholamines were all potentiated by ouabain, a potent inhibitor of Na, K-ATPase. Omission of K from the medium, a condition which suppresses the Na, K-ATPase activity, also augmented these cell responses caused by veratridine. On the contrary, diphenylhydantoin, which is known to decrease the intracellular concentration of Na, reduced the veratridine-induced influx of ²²Na, ⁴⁵Ca and secretion of catecholamines. The potentiating effects of ouabain on the veratridine-induced cell responses were all abolished by diphenylhydantoin. These findings imply that veratridine, ouabain and K removal as well as diphenylhydantoin modulate the intracellular accumulation of ²²Na which is involved in the influx of ⁴⁵Ca and the secretion of catecholamines.     

On the release of catecholamines and dopamine-beta-hydroxylase evoked by ouabain in the perfused cat adrenal gland.

1 Secretion of catecholamines (CA) and dopamine-beta-hydroxylase (DBH) activity from the retrogradely perfused cat adrenal gland was studied following ouabain infusion. Perfusion with ouabain (10(-4) M) for 10 min caused a gradual release of CA in the effluent which reached its peak 30 min after the ouabain pulse, and was maintained constant for at least 1 h. The effect of ouabain seemed to be irreversible. 2 Mecamylamine, while blocking the CA secretory effects of acetylcholine (ACh) perfusion, did not affect the secretion of CA evoked by ouabain. In denervated adrenal glands, ouabain-induced CA secretion was similar to that in the contralateral, innervated gland. However, physostigmine perfusion potentiated the CA secretory effects of ouabain. 3 The release of CA evoked by ouabain was accompanied by a proportional release of DBH activity. The time course of appearance of DBH activity followed the pattern of CA release. 4 The CA and DBH outputs in response to a pulse of ouabain were suppressed in the absence of calcium. Calcium reintroduction to a calcium-free perfused, ouabain-treated gland not only restored but greatly potentiated the release of CA and DBH. The amplitude of the secretory response to calcium reintroduction in ouabain-treated glands was proportional to the extracellular calcium concentration, and was antagonized by an external sodium-deficient medium. 5 These data demonstrate that ouabain releases CA from the perfused cat adrenal gland by a calcium-dependent exocytotic mechanism. The secretory effect of ouabain is not secondary to the release of ACh from cholinergic nerve terminals present in the adrenal gland, but due to a direct action on the chromaffin cell itself. In addition, the results suggest that this action is exerted through redistribution of monovalent cations secondary to the inhibition by the glycoside of the sodium pump. Such monovalent cation redistribution may cause a rise of intracellular ionized calcium levels through the activation of an internal sodium-dependent calcium influx system probably located in the chromaffin cell membrane.     

The modification of inotropic action of ouabain by 6-hydroxydopamine and alpha-methyl-p-tyrosine in dogs.

The action of ouabain on myocardial inotropism pretreated with 6-hydroxydopamine and alpha-methyl-p-tyrosine was studied. Inotropic action of ouabain was not changed by depletion of catecholamines in the brain, in which the central sympathetic neurons were destroyed by an intraventricular administration of 6-hydroxydopamine. Systemic administration of 6-hydroxydopamine and alpha-methyl-p-tyrosine reduced ventricular catecholamines to 5.3% and 20.8% of the control, respectively. Percent increase of contractility by ouabain after the pretreatment of 6-hydroxydopamine and alpha-methyl-p-tyrosine was reduced to 16.7% and 50.3% of the control, respectively. The results obtained suggest that catecholamines in the myocardium play some important role in producing cardiotonic action of cardiac glycosides. Brain catecholamines or the central sympathetic nervous system do not appear to participate in the exertion of the positive inotropic action of cardiac glycosides.     

Influence of panaxatriol-type saponin on secretion of catecholamines from isolated perfused rabbit adrenal gland

In the previous observations, it was reported that both total ginseng saponin and panaxadiol revealed the marked secretory effect of catecholamines (CA) from the rabbit adrenal gland and that CA secretion induced by them is due to dual mechanisms, cholinergic action and the direct action. In the present study, an attempt to investigate the effect of panaxatriol-type saponin (PT), which is known as an active component of Korean ginseng, on the secretion of CA from the rabbit adrenal gland was made. PT (200 μg) administered into adrenal vein evoked significantly secretion of CA from the isolated perfused rabbit adrenal gland. Secretory effect of CA produced by PT was attenuated clearly by treatment with chlorisondamine or adenosine, but was markedly increased by physostigmine. Perfusion of Krebs solution containing PT (200 μg) for 30 min potentiated greatly secretion of CA induced by acetylcholine. PT-induced CA secretion was weakened considerably by ouabain treatement or perfusion of calcium-free Krebs solution. These experimental data demonstrate that PT releases CA from the isolated perfused rabbit adrenal gland by a calcium-dependent exocytotic mechanism. It seems that the secretory effect of PT is caused through the release of acetylcholine from cholinergic terminals present in the adrenal gland and a direct action on the chromaffin cell itself.     

Phenylalanine, brain phenethylamine and motor activity in the rat

Varying doses of L-phenylalanine were administered intraperitoneally to rats and motor activity determined by means of photocells in an activity cage. Control rats, and rats whose brain catecholamines were depleted by reserpine, α-methyl-p-tyrosine, or intraventricular 6-hydroxydopamine showed a decrease or no change in motor activity following phenylalanine injections. Injections of the decarboxylase inhibitor, Ro 4–4602, failed to alter the effect of phenylalanine. After injections of [³ H]phenylalanine, [³ H] β-phenethylamine accounted for less than 5 % of brain tritium and radioactivity associated with this amine declined rapidly with time. The proportion of brain radioactivity due to β-phenethylamine was increased by the monoamine oxidase inhibitor, pargyline, and was not affected by the peripheral decarboxylase inhibitor Ro 4–4602. When Ro 4–4602 and pargyline were given together, the increase in radioactive phenethylamine produced by pargyline was prevented, suggesting that a significant proportion of brain phenethylamine may be synthesized in the periphery and can enter brain if monoamine oxidase is inhibited.     

Long-term regulation of catecholamine formation by ouabain in cultured bovine adrenal chromaffin cells

The long-term effects of ouabain, an inhibitor of Na+/K+ -ATPase, on catecholamine formation in cultured bovine adrenal chromaffin cells were examined. The increase in [14C]catecholamine formation from [14C]tyrosine induced by ouabain was dependent on incubation time, and its maximal effect was observed after incubation for 8 h. The stimulatory effect of ouabain was concentration dependent (10-300 nM), causing maximal stimulation at 300 nM. The formation of [14C]catecholamines induced by ouabain was not increased by incubation with [14C]DOPA instead of [14C]tyrosine. Ouabain-induced [14C]catecholamine formation was influenced by decreases in extracellular Ca2+ concentration, but not by the presence of cycloheximide or actinomycin D. These results suggested that ouabain stimulates continuous activation of hydroxylation of tyrosine through a Ca2+ -dependent mechanism in cultured bovine adrenal chromaffin cells.     

Ionic and metabolic requirements for stimulation of secretion by ouabain in bovine adrenal medullary cells

The effect of ouabain on the secretion of catecholamines from isolated bovine adrenal medullary cells was investigated. Ouabain enhances the basal rate of secretion approximately 2-fold, with half-maximal stimulation occurring at a glycoside concentration of around 5 X 10(-7) M. Parallel measurements of the release of dopamine beta-hydroxylase (EC 1.14.17.1) (an enzyme associated with chromaffin granules) and lactate dehydrogenase (EC 1.1.1.27) (which is confined to the cytosolic compartment) suggest that this increase in secretion occurs as a result of an enhanced rate of exocytosis rather than by any other route. The stimulatory effect of ouabain is dependent on extracellular sodium but is maintained in the nominal absence of calcium and is unaffected by changes in the major external anion. Neither tetrodotoxin nor phenoxybenzamine alters the response to glycoside treatment, but the calcium channel blocker methoxyverapamil reduces the catecholamine secretion evoked by ouabain in a dose-dependent fashion. This study serves to characterize the secretory action of ouabain in isolated chromaffin cells and to provide a foundation for the ion flux studies reported in the following paper.     

The effect of catecholamines on the influx of calcium and the development of tension in denervated mouse diaphragm muscle.

1 The nature of the catecholamine-induced contracture of chronically denervated mouse diaphragm muscle has been investigated and compared with the contractural response evoked by acetylcholine. 2 The time course of onset of catecholamine-sensitivity in denervated diaphragm muscles was similar to the development of acetylcholine sensitivity. However, catecholamine contractures were absent in tissues denervated for periods longer than 90 days whereas acetylcholine-sensitivity was still evident several months after denervation. 3 The catecholamine-induced contracture of the denervated muscle was inhibited specifically by beta-receptor blocking drugs and was unaffected by alpha-receptor blocking drugs and cholinoceptor antagonists. 4 Catecholamine-induced contractures of denervated muscles, unlike contractures to acetylcholine, were dependent upon the presence of spontaneous fibrillation and the amplitude of spontaneous fibrillation was increased by catecholamines. Fibrillation was absent in the presence of tetrodotoxin (1 muM), 2,4-dinitrophenol (10 muM), potassium cyanide (10 muM), ouabain (100 muM), in lithium chloride Ringer solution and at low temperature. Under these conditions catecholamine-induced contractures, but not those to acetylcholine, were abolished. 5 Labelled calcium was found progressively to enter denervated muscle fibres and this entry of calcium was increased by catecholamines. It is suggested that this calcium entry may represent either an increased calcium permeability of denervated muscle fibres which is increased further by catecholamines or the presence of a calcium current that occurs during the fibrillatory potentials of denervated muscle.     

d-Amphetamine-induced hepatotoxicity: possible contribution of catecholamines and hyperthermia to the effect studied in isolated rat hepatocytes

Amphetamines are indirect-acting sympathomimetic drugs widely abused due to their physical and psychostimulating effects. However, the use of these drugs has been associated with numerous reports of hepatotoxicity. While glutathione depletion induced by amphetamines contributes to the exposure of hepatocytes to oxidative damage, other indirect effects attributed to amphetamines may have a role in cell injury. To examine this possibility, Wistar rats were used for plasma measurements of d-amphetamine and catecholamines (noradrenaline, adrenaline and dopamine) (15 min) after i.p. injection of d-amphetamine (5, 20 and 80 mg/kg). Freshly isolated rat hepatocytes were put into contact for 2 h with concentrations of d-amphetamine and catecholamines similar to those found in vivo. Since hyperthermia is a common consequence of acute amphetamine intake, the study using isolated hepatocytes was conducted at 37 °C and also at 41 °C in order to simulate high temperature levels. We found that hyperthermia was an important cause of cell toxicity: in vitro, a rise in incubation temperature from 37 to 41 °C causes oxidative stress in freshly isolated rat hepatocytes, as shown by a depletion of reduced glutathione (GSH; 23%), an increase of oxidized glutathione (GSSG; 157%), the induction of lipid peroxidation with 77% increase of thiobarbituric acid substances TBARS) and the consequent loss of cell viability (≤ 44%). Single treatment of isolated hepatocytes with catecholamines at 37 °C induced lipid peroxidation (29% increase of TBARS) but had no effect on glutathione or cell viability. Conversely, a single treatment with d-amphetamine induced glutathione depletion (≤ 24% depletion of GSH) with no effect on lipid peroxidation or cell viability. Also, d-amphetamine potentiated the induction by catecholamines of lipid peroxidation at 37 °C (≤ 48% increase of TBARS), while concomitant treatment of d-amphetamine and catecholamines potentiated cell death at 41 °C (≤ 56% of cell death) although no effect on viability was seen at 37 °C. It is concluded that the aforementioned modifications induced by d-amphetamine in vivo are cytotoxic to freshly isolated rat hepatocytes. Received: 30 October 1996 / Accepted: 13 January 1997     

The lack of effect of oxytetracycline on responses to sympathetic nerve stimulation and catecholamines in vascular tissue.

The effects of oxytetracycline, an inhibitor of amine binding in connective tissue, on the responses of perfused rabbit ear arteries to sympathetic nerve stimulation and to intraluminally administered noradrenaline were examined. The contractions of aortic strips to catecholamines in the presence of oxytetracycline were also examined. Oxytetracycline (0.1 mM) had no discernable effect on the magnitude of constrictions, measured as reductions in flow, produced by either nerve stimulation (0.5-10 Hz) or noradrenaline (0.5-50 ng) in the ear artery. In addition, the time taken for vessels to recover towards control flow values after endogenously released or exogenously applied noradrenaline had acted was not increased by oxytetracycline. Oxytetracycline (0.1 mM) did not alter the position or shape of the concentration-response curve to noradrenaline nor did it enhance the amplitude of individual responses to catecholamines in aortic strips. It is concluded, contrary to the observations of Powis (1973), that oxytetracycline does not increase the magnitude or duration of responses to sympathetic nerve activation or to catecholamines and that binding to connective tissue is of no material consequence in terminating their action in vascular tissue.     

The effects of acute and chronic administration of morphine on the turnover of brain and adrenal catecholamines in rats

Brain and adrenal catecholamine turnover in adult female rats treated with morphine was investigated. A different time course response of brain and adrenal catecholamines to -methyl-p-tyrosine methylester (AMT) administration in normal rats was observed; the catecholamine turnover rate in adrenal glands appeared to be much slower than in the brain. Acute morphine increased the turnover of brain dopamine and noradrenaline as well as of adrenal catecholamines, whereas chronic morphine treatment induced a decrease in the turnover of brain noradrenaline. Withdrawal induced by nalorphine produced an increase in the utilization of brain noradrenaline and adrenal catecholamines; this effect could be related to the withdrawal stress situation induced by the opiate antagonist. Although the mechanism of morphine action may implicate other neurotransmitters besides catecholamines, our results contribute to evidence that brain and adrenal catecholamines could be involved in the mechanism of morphine tolerance and/or dependence.     

Catecholamine release evoked by lithium from the perfused adrenal gland of the cat.

The effect of Li on catecholamine release by cat isolated retrogradely perfused adrenal gland was investigated. Replacement of Na (119 mM) by Li in the Krebs solution evoked a progressive increase in the spontaneous release of catecholamines that reached a maximum within 45 min and was Ca-dependent. This response was specific for Li, since sucrose or choline used as osmotic substitutes for Na, failed to increase the spontaneous release of catecholamines by the adrenal gland. In glands perfused with Li-Krebs for 30 min a sharp secretory response was observed when Li was replaced by sucrose or choline; no such an effect was seen when Li was replaced by Na. Partial replacement of Na by sucrose, in ouabain (10(-4) M, 10 min) pretreated glands perfused with normal Krebs induced a sharp increase in the catecholamine output whilst replacement by Li produced a significantly lower response. Reintroduction of Ca (2.5 mM, 2 min) in glands previously perfused with Ca-free, Mg-containing Li-Krebs, evoked a sharp increase in catecholamine release. No such an effect was seen when the glands were perfused with Ca-free normal, choline- or sucrose-Krebs. The release of catecholamines evoked by Ca reintroduction in glands previously perfused with Ca-free Li-Krebs was directly dependent on the Li concentration and the length of time of the Li loading period. In summary, our results indicate that Li accumulates in the cells and can partially substitute Na in the Na-Ca counter-transport system at the plasma membrane of the chromaffin cell.     

Increases in plasma cyclic AMP dependent on endogenous catecholamines.

Administration of tyramine (with or without phentolamine) as well as induction of ether anesthesia or insulin hypoglycemia caused a sharp increase in plasma cyclic AMP in rats. Based on the findings that the treatment of rats with reserpine, 6-hydroxydopamine, cocaine or propranolol totally abolished tyramine-induced increases in plasma cyclic AMP, it was concluded that catecholamines released from sympathetic neuronal terminals by tyramine could activate adenylate cyclase via the stimulation of postsynaptic beta-adrenoceptors. In contrast, catecholamines secreted from adrenal medulla were largely responsible for the increase in plasma cyclic AMP induced by ether anesthesia; whereas glucagon, in addition to adrenal catecholamines, played a significant role in hypoglycemia-induced increases in plasma cyclic AMP. Assay of plasma cyclic AMP following these stimuli is very promising as a test for adrenergic activities in experimental and clinical studies.     

Nicotine-induced release of catecholamines from rat hippocampus and striatum

The present report is a comparative study of [³H]catecholamine release from noradrenergic and dopaminergic neuron terminals of the central nervous system, induced by nicotine and potassium depolarization. Striatal and hippocampal slices of rat brain that had incorporated [³H]catecholamines in previous incubations with the radioamines were superfused and stimulated by nicotine and high potassium. Nicotine produced a marked [³H]catecholamine release from these two brain areas, this effect being much greater in the striatum. The time course of radioactive efflux released by nicotine was different from that induced by high potassium and similar to that evoked by tyramine. Nicotine induced released was dose dependent, inhibited by low temperature, independent of extracellular calcium and not inhibited by an excess of magnesium. Studies with newly synthesized [³H]dopamine and [¹⁴C]urea suggested that nicotine acted on the storage vesicles. [³H]catecholamine release was continuous during prolonged nicotine stimulation, in contrast to the transient efflux induced by prolonged stimulation by potassium, and additive when the slices were superfused simultaneously with nicotine plus potassium. Previous nicotine superfusion did not modify the typical potassium release, and the response to nicotine was also not altered when the slices were exposed previously to high potassium. Based on these results, the mechanism of nicotine release of catecholamines from central catecholaminergic neuron terminals is discussed.     

Effects of adrenergic drugs on pressor responses to hypothalamic stimulation

Summary In 12 female dogs renal excretion and catabolism of ¹⁴C-(±)-adrenaline, ¹⁴C-(±)-noradrenaline, ¹⁴C-dopamine and ³H-(±)-normetanephrine were investigated using a modified stop-flow technique. Radioactive compounds were infused, together with inulin, into the left renal artery for 10 min. During the first 2 min of the infusion period the left ureter was occluded. Urine samples were serially collected from both kidneys up to the end of the infusion. In the urine the total radioactivity and the pattern of radioactive metabolites were measured.On average, the infused kidney excreted from the infused dose of ¹⁴C-adrenaline 9.4% as adrenaline, 27.9% as metanephrine and 5.8% as deaminated or conjugated metabolites. From infused ¹⁴C-noradrenaline 7.4% was excreted as noradrenaline, 3.5% as normetanephrine and 1% as deaminated or conjugated compounds. When ³H-normetanephrine was infused the urine contained only radioactive normetanephrine (22.2%). From the infused dose of ¹⁴C-dopamine 9.6% was excreted as dopamine, 16.2% as 3-O-methyldopamine and 3.7% as deaminated or conjugated compounds. — Urine from the other kidney contained 1/25 to 1/5 the radioactivity of that from the infused side, but the pattern of radioactive compounds was similar.From the excretion rate of simultaneously infused inulin the filtration fraction of the infused kidney was determined. That part of the infused ¹⁴C-catecholamines which was excreted unmetabolized in the urine, corresponds to the filtration fraction in this kidney. Therefore, it is suggested, that in mammals the unmetabolized catecholamines of the urine are mainly excreted by glomerular filtration and not by tubular secretion. On the other hand, the urinary O-methylated radioactive catecholamines, which were excreted by the infused kidney at a high rate, were formed in this organ from the infused catecholamines and were excreted by tubular secretion. Thus, in mammals tubular secretion is linked to an inactivation of these compounds by O-methylation.Supported by the Deutsche Forschungsgemeinschaft.     

Anaphylaxis-induced increase in plasma cyclic AMP dependent on endogenous catecholamines in rats

Plasma cyclic AMP levels during anaphylactic shock in rats was studied in 7 groups of animals: (1) control rats; (2) rats with adrenomedullectomy; (3) rats treated with propranolol; (4) rats with reserpinization; (5) rats with 6-hydroxydopamine-induced chemical sympathectomy; (6) rats treated with hexamethonium, and (7) rats treated with cocaine (catecholamine uptake inhibitor). All experiments were carried out in ovalbumin-sensitized rats. Plasma cyclic AMP showed a rapid increase during anaphylactic shock in control rats. Adrenomedullectomy abolished the anaphylaxis-induced increase in plasma cyclic AMP, while hexamethonium had no effect. Propranolol caused a dose-dependent abolition of the increase. The treatment of rats with reserpine, 6-hydroxydopamine or cocaine partially inhibited the increase in plasma cyclic AMP. The results show that the adrenal medulla is the major source of catecholamines during anaphylactic shock, and that catecholamines in the adrenergic neuronal terminals may be partly responsible for the anaphylaxis-induced increase in plasma cyclic AMP.     

Chronic administration of morphine in cats: effects on adrenal and urinary catecholamines.

Adrenaline and noradrenaline levels in the adrenal glands and the excretion of both bioamines in urine of adult cats were investigated after chronic administration of morphine and nalorphine-induced withdrawal. After 7 days of daily consecutive morphine treatment, a significant increase in the adrenal noradrenaline content and a drop in adrenaline content were observed. After 2 weeks of daily injection of morphine, no significant changes were observed in the adrenal catecholamine level. One month of treatment with the opioid caused a significant increase in the adrenal content of both adrenaline and noradrenaline. Urinary excretion of catecholamines was significantly increased during the 4 weeks of treatment. In animals subjected to spontaneous or induced withdrawal with nalorphine, the adrenal content of catecholamines was altered and the ratio adrenaline/noradrenaline in the adrenal gland was shifted towards noradrenaline. A first injection of morphine produced an excitant manic response characterized by hyperexcitement and aggressive behaviour; animals chronically treated with the drug showed a progressively diminished response to this effect of the drug. It is concluded that physical dependence on morphine is reached by cats chronically treated with morphine and that this effect of the drug influences adrenomedulllary function in a different fashion depending on the stage of morphine treatment.     

Clozapine-induced myocarditis: role of catecholamines in a murine model

Clozapine, an atypical antipsychotic, is very effective in the treatment of resistant schizophrenia. However, cardiotoxicity of clozapine, particularly in young patients, has raised concerns about its safety. Increased catecholamines have been postulated to trigger an inflammatory response resulting in myocarditis, dilated cardiomyopathy, and death, although this has not yet been thoroughly studied. Here, we used the mouse to study whether clozapine administration could cause adverse myocarditis associated with an increase in catecholamines. Male Balb/C mice, age ~6 weeks, were administered 5, 10 or 25 mg/kg clozapine daily for 7 and 14 days; one group was administered 25 mg/kg clozapine plus 2 mg/kg propranolol for 14 days. Saline-treated mice served as controls. Heart sections were stained with hematoxylin and eosin for histopathological examination. Plasma catecholamines were measured with HPLC. Myocardial TNF-alpha concentrations were determined by ELISA. Histopathology of clozapine-treated mice showed a significant dose-related increase in myocardial inflammation that correlated with plasma catecholamine levels and release of TNF-alpha. Propranolol significantly attenuated these effects. A hypercatecholaminergic state induced by clozapine could explain the occurrence of myocarditis in some patients. Our data suggest that a beta-adrenergic blocking agent may be effective in reducing the incidence and severity of clozapine-induced myocarditis.