Beta-adrenoceptor Desensitization Research Articles

Cytosolic factor(s) determines the impaired desensitization of beta-adrenoceptors in the myocardium of spontaneously hypertensive rats

Loss of membrane-bound β-receptors in the myocardium of spontaneously hypertensive rats has been explained on the basis of enhanced agonist-induced desensitization. However, in both cardiac myocytes and cell-free preparations, isoproterenol and cyclic AMP-dependent protein kinase are less effective in desensitizing β-receptors from hypertensive compared to normotensive animals. Reconstitution of cardiac membrances from hypertensive rats with cytosol from normotensive controls restores the ability of isoproterenol and protein kinase A to desensitize β-receptors. Impaired desensitization in spontaneously hypertensive rats is, therefore, not due to intrinsic defects of the β-receptor but, rather, to absence of regulatory cytosolic factor(s).

A Different Desensitization Pattern of Cardiac β-Adrenoceptor Subtypes by Prolonged In Vivo Infusion of Isoprenaline

(-)Isoprenaline was continuously administered to rats at a rate of 0.4 mg/kg/h for 7 days via subcutaneously (s.c.) implanted osmotic minipumps. This treatment induced cardiac hypertrophy and a marked decrease in basal as well as catecholamine-stimulated adenylate cyclase activity in a ventricular plasma membrane fraction. The total number of beta-adrenoceptors was downregulated by one-half the amount of the receptor sites obtained in a control group. However, in the isoprenaline-treated group, the beta 2-adrenoceptors constituted a significantly smaller proportion of the total beta-adrenoceptor population (28%) than in the control group (50%). Transformation of these relative into absolute values indicates that prolonged isoprenaline treatment induced a significantly higher downregulation of beta 2- than of beta 1-adrenoceptors. The fact of a different beta-adrenoceptor desensitization pattern in response to in vivo administration of nonselective beta-adrenergic agonists therefore must be taken into consideration when desensitization is used as a method for determination of subtype selectivity of an agonist per se. However, we were unable to detect the "lost" beta-adrenoceptors in a light vesicular fraction. In our study, this fraction was not separable from plasma membranes, as substantiated by levels of plasma membrane markers as high as in the plasma membrane fraction and by a guanine nucleotide-dependent adenylate cyclase activity.

Catecholamine-induced desensitization of brain beta adrenoceptors in vivo and reversal by corticosterone

The function of beta adrenoceptors in the brain was studied with a new technique involving the microdialysis collection of extracellular cyclic AMP (cAMP). Prolonged infusion of isoproterenol (2 hr) was found to induce a marked desensitization of the beta receptor-cAMP response. Administration of corticosterone (10 mg/kg) 3 hrs prior to the infusion markedly attenuated the desensitization. The results indicate that the catecholamine-induced desensitization of brain beta adrenoceptors can be studied in vivo and that corticosterone is capable of reversing this effect in the brain as has been reported previously for peripheral tissues.

Impaired desensitization of cardiac beta-adrenoceptors in the spontaneously hypertensive rat.

Reduced inotropic responsiveness by spontaneously hypertensive rats (SHR) to beta agonists is associated with, and may be secondary to, a decline in cardiac membrane-bound beta adrenoceptors. It is widely assumed that this decline reflects a desensitization of the receptor secondary to enhanced sympathetic drive, but direct evidence is lacking. We examined this issue in cell-free cardiac preparations from age-matched SHR and Wistar-Kyoto (WKY) rats. Isoproterenol induced a time- and concentration-dependent decline in membrane-bound beta receptors but was considerably more effective in WKY than in SHR (43% decline versus 29% in SHR, P less than 0.01). Since cyclic AMP-protein kinase inhibitor prevented only 15-20% of the isoproterenol-induced decline, this was probably mediated by the beta receptor kinase. Cyclic AMP-dependent protein kinase induced a 38% desensitization in WKY rats but was completely ineffective in SHR. These results suggest that desensitization of the beta receptor is impaired in SHR, probably because the structural organization of the receptor in the cardiac membranes is altered.

Impaired response of experimental diabetic mice to tricyclics: A possible beta-adrenergic mechanism

Diabetes is reportedly associated with alterations in peripheral and central noradrenergic systems. The latter might be involved in the antidepressant effects of imipramine-like drugs in both humans and animals. Therefore, it is possible that diabetics show an impaired responsiveness to tricyclics. To test this possibility the effects of streptozotocin (STZ)-induced experimental diabetes in mice were assessed in two psychopharmacological tests: 1) the reversal of apomorphine- (16 mg/kg) induced hypothermia and 2) the hypoactivity by a direct beta-agonist (clenbuterol 0.06 mg/kg). At day 15 after STZ or vehicle treatment, imipramine (4 mg/kg) antagonized the apomorphine-induced hypothermia in diabetic (D) and nondiabetic (ND) mice and clenbuterol produced hypoactivity in both groups. At day 30 and 45, the ability of imipramine (1, 2, 4, 8, 16 mg/kg), clomipramine (8 mg/kg) and desipramine (2 mg/kg) to reverse apomorphine-induced hypothermia disappeared at the same time that clenbuterol lost its ability to induce hypomotility in D mice. These impaired responses on both tests were corrected by a short period of insulin therapy. These two tests may reflect central beta- adrenergic functions. Therefore, these data suggest that the impaired responsiveness of diabetic mice might be due at least in part to a noradrenergic dysfunction. Possibly, in diabetes, a beta-adrenoceptor desensitization identical to that observed at the peripheral level occurs in the central nervous system. The possibility that a thyroid hormone deficiency may be involved was also tested. Decreased T3 plasma levels were found in D mice concomitant with the impaired pharmacological responses and T3 supplementation turned these responses to normal. An hypothetical link between these thyroid effects and the beta-adrenergic desensitization in D mice could be suggested but remains to be determined.

Interaction of some atypical antidepressants and adrenoceptor antagonists with imipramine: A behavioural study with isoprenaline-induced drinking

The systemic administration of isoprenaline to rats produced a dose-dependent increase in drinking which was antagonized by propranolol. While oral administration of the antidepressant, imipramine, alone had no significant effect on this response, the increase was significantly inhibited by administration of imipramine together with each of the following drugs over a period of 4 days: bupropion (21.0 mg/kg/day, p.o.), a selective inhibitor of the uptake of dopamine and nomifensine (10.6 mg/kg/day, p.o.), a relatively selective dopamine and a blocker of the uptake of noradrenaline. Similarly, the combination of the selective alpha 1 -adrenoceptor antagonist, prazosin (2.37 mg/kg/day, p.o.); the selective alpha 2-adrenoceptor antagonist, yohimbine (2.38 mg/kg/day, p.o.) or the non-selective alpha-adrenoceptor blocker, phentolamine (4.65 mg/kg/day, p.o.), with imipramine caused a significant inhibition of the isoprenaline-induced drinking. It is concluded that fast desensitization of central beta-adrenoceptors in the rat can be produced after the oral subacute simultaneous administration of imipramine with alpha-adrenoceptor antagonists or atypical antidepressants, such as nomifensine or bupropion.

Acidic sphingomyelinase: Relationship with antidepressant-induced desensitization of beta-adrenoceptors

Previous results indicate a dose-dependent decrease of lysosomal sphingomyelinase activity induced by tricyclic antidepressants in cell cultures. A possible association of this effect with the antidepressant-induced down-regulation of beta-adrenoceptors was postulated. We report here the determination of beta-adrenoceptor binding sites and lysosomal sphingomyelinase activity in the cerebral cortex of rats treated chronically with desipramine or with the potential antidepressant drug midalcipran (which is devoid of effect on beta-adrenoceptors). The effect of midalcipran on lysosomal sphingomyelinase activity was also determined on C6 glioma cells. In C6 glioma cells, midalcipran did not decrease sphingomyelinase activity, at variance with the enzymatic inhibition induced by desipramine (DMI). In the rat cerebral cortex, neither DMI nor midalcipran modified sphingomyelinase activity. In agreement with previously reported effects, DMI induced beta-adrenoceptor desensitization in the rat cerebral cortex, while midalcipran remained ineffective. Our results indicate that in the rat cerebral cortex, the activity of lysosomal sphingomyelinase is not modulated by chronic treatment with antidepressant drugs, whatever their effect on beta-adrenoceptor sites. Our results suggest that sphingomyelinase activity is not associated with the desensitization of beta-adrenoceptors, taken as an index of the therapeutic action of antidepressants. The results indicate that care should be taken when extrapolating to in vivo situations the conclusions derived from cell culture conditions.

Mechanisms regulating the adrenaline-induced long-term potentiation in bullfrog sympathetic ganglia.

Two regulatory mechanisms on the long-term potentiation of transmitter release induced by adrenaline (adr.-l.t.p.) in bullfrog sympathetic ganglia were studied by recording intracellularly the fast excitatory postsynaptic potentials. An increase in exposure time to adrenaline from 10 min to 60 min did not enhance the magnitude of adr.-l.t.p. However, increasing an exposure time to dibutyryl cyclic AMP (1 mM) up to 60 min progressively enhanced the magnitude of the nucleotide-induced potentiation, indicating the desensitization of the beta-adrenoceptor. The desensitization remained at 20 min after the removal of adrenaline in all the five cells but disappeared at 60-90 min in four cells out of eight. Under the latter condition, the second l.t.p. was summated on the first one. Dibutyryl cyclic GMP (100 microM) blocked the generation of the l.t.p. induced by dibutyryl cyclic AMP (1 mM) as well as that of adr.-l.t.p. Muscarine (10 microM) or adenosine (1 mM), a possible candidate for raising intraterminal cyclic GMP, did not significantly affect adr.-l.t.p. These results suggest that adr.-l.t.p. is regulated by the desensitization of beta-adrenoceptor and a process which involves endogenous cyclic GMP acting on a step subsequent to the cyclic AMP production.

Studies in vivo and in vitro of terbutaline-induced beta-adrenoceptor desensitization in healthy subjects.

Beta-Adrenoceptor function was studied in eight healthy subjects before, during and 24 and 72 h after cessation of 2 weeks continuous oral treatment with the beta 2-adrenoceptor agonist terbutaline (sustained release, 7.5 mg twice daily). In vivo, blood pressure, heart rate, plasma noradrenaline and plasma cyclic AMP responses to isoprenaline (0.01, 0.02 and 0.05 microgram min-1 kg-1 intravenously) were related to the plasma concentrations of isoprenaline. for comparison, beta 2-adrenoceptor function was evaluated in lymphocytes in vitro by studies of isoprenaline-induced accumulation of cyclic AMP and radioligand binding studies using 125I-iodohydroxybenzylpindolol. In vivo, the beta 2-mediated plasma cyclic AMP response to isoprenaline was markedly attenuated during terbutaline treatment and was still reduced by 38% (P less than 0.05) 72 h after discontinuation of treatment. The blood pressure and heart rate responses to isoprenaline were unaffected by treatment. Isoprenaline-induced elevations of plasma noradrenaline concentrations were markedly reduced during terbutaline treatment. This indicates an attenuation of isoprenaline-induced increases in sympathetic nerve function and could explain why no attenuation of the isoprenaline-induced vasodilatation was observed. Thus, plasma cyclic AMP seems to be a better marker than diastolic blood pressure when evaluating beta 2-adrenoceptor responsiveness in vivo in man, since it is not influenced by counter-regulatory increases in sympathetic nerve activity and/or noradrenaline overflow from sympathetic nerves. In lymphocytes, the isoprenaline-stimulated cyclic AMP accumulation was reduced by 75% and the beta-adrenoceptor binding sites were reduced by 40% 12 h after dosing. Also the lymphocyte beta 2-adrenoceptors recovered slowly after withdrawal of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Endotoxin-induced alterations in guinea pig coronary vascular beta 2-adrenoceptor function: a role for cyclo-oxygenase products.

Bordetella pertussis and in particular its cell wall constituent endotoxin induces a hyporesponsiveness of the coronary vascular beta2-adrenoceptor and a hyperreactivity of the alpha2-adrenoceptor in the guinea pig coronary vascular system [5]. In literature, much attention has been paid to betaadrenoceptor desensitization and several hypotheses have been proposed in order to explain the molecular mechanisms of this phenomenon [4]. The involvement of mediators other than catecholamines has been considered. Among these mediators, arachidonic acid and its metabolites have been suggested to play a role in desensitization of beta-adrenoceptors in various experimental designs [2, 3, 4]. Since endotoxin is a powerful stimulator of the immune system and thus initiates the biosynthesis of various inflammatory mediators among which are the arachidonic acid metabolites, it seems of particular interest to examine the hypothesis that arachidonic acid metabolites are involved in beta2-adrenoceptor impairment in the coronary vascular system of the guinea pig heart after endotoxin. Furthermore, these experiments might also elucidate the possible role of prostaglandin synthesis for alpha2-adrenoceptor hyperreactivity.

Concomitant glucocorticoid treatment prevents the development of beta-adrenoceptor desensitization in the guinea pig lung.

The aim of the present study was to investigate, whether concomitant administration of the synthetic glucocorticoid betamethasone (BM), theophylline (THEO), or the muscarinic antagonist ipratropium bromide (IPRA) could influence the desensitization-associated decrease of beta-adrenoceptors in the guinea pig lung during prolonged in vivo treatment with the beta 2-agonist terbutaline (TER). The animals were sacrificed 20 hrs after the last drug dosage and the lung membrane homogenates were prepared for 3H-dihydroalprenolol (3H-DHA) binding in vitro. Treatment with TER 200 micrograms/kg subcutaneously twice a day for five days decreased by 22% the maximum number of binding sites (Bmax) at saturation in comparison with the saline-treated controls. Concomitant administration of BM 2 mg/kg intraperitoneally abolished this effect of TER, whereas THEO 20 mg/kg or IPRA 5 micrograms/kg failed to modify it. None of the in vivo treatments affected the binding affinity of 3H-DHA. In vitro, TER inhibited in a concentration-dependent manner 3H-DHA binding to the lung membranes of untreated guinea pigs. At high concentrations IPRA, but not THEO or BM, showed some binding to the beta-receptors as well. Thus, it is concluded that glucocorticoids may prevent beta-adrenoceptor desensitization in the lungs via an indirect mechanism, e.g. inhibition of phospholipase A2 enzyme.

Cardiac beta-adrenoceptor desensitization after sinoaortic baroreceptors denervation or isoproterenol-pretreatment

The chronotropic reactivity to beta-adrenergic stimulation was studied in isolated perfused hearts from sinoaortic baroreceptors denervated (SAD) rats in the phases of maximal tachycardia (5 h) and normalized heart rate (15 days) or after repeated injection of isoproterenol (1 mg/kg, 3 times daily for 10 days). The threshold doses of isoproterenol needed to cause a significant acceleration of heart rate were significantly higher in acute SAD (1.6 X 10(-7)M), chronic SAD (3.2 X 10(-7)M), and isoproterenol-pretreated (6.4 X 10(-7)M) rats than in control rats (0.8 X 10(-7)M). The maximal increment in basal heart rate was lower in acute (59%) and chronic (48%) SAD rats and in isoproterenol-pretreated rats (26%) than in control rats (78%). These results suggest that the increased efferent sympathetic outflow to the heart induced by SAD causes desensitization of cardiac beta-adrenoceptors and may explain the gradual tachycardia disappearance in this SAD hypertensive rats.

Role of cyclic AMP in cardiac beta-adrenoceptor desensitization: studies using prenalterol and inhibitors of phosphodiesterase.

We examined the effects of prolonged exposure of cardiac cells in primary culture to the partial beta-adrenoceptor agonist prenalterol and inhibitors of phosphodiesterase on their subsequent ability to increase intracellular cyclic AMP during a 5-min exposure to 50 microM isoprenaline (receptor responsiveness). Although prenalterol possesses only 7% of the agonist activity of isoprenaline on adenylate cyclase, it induces extensive beta-adrenoceptor desensitization. Three hours after exposing the cells to 1 microM prenalterol, beta-adrenoceptor responsiveness was reduced by 40% (p less than 0.05), whereas after 12 h the reduction averaged 55%. Prolonging the incubation time to 48 h had no further effect on the magnitude of receptor desensitization. The magnitude of the desensitization was concentration dependent. On exposure of cells to 10(-8) M prenalterol for 16 h, receptor responsiveness was reduced by 19%, and at concentrations of 1 microM and higher responsiveness was reduced by 60% (p less than 0.01). Receptor desensitization appeared to be due to an inability of receptors to activate adenylate cyclase as well as to receptor loss. To investigate if beta-adrenoceptor desensitization as well as receptor loss could be mediated by cyclic AMP, the cells were exposed for 16 h to inhibitors of phosphodiesterase. Exposure of cells to the phosphodiesterase inhibitor isobutylmethylxanthine (0.1 mM) (which increased intracellular cyclic AMP by between 50 and 150%) also induced receptor desensitization. The reduction in receptor responsiveness averaged 62% (p less than 0.01). The loss in responsiveness could be accounted for by an inability of receptors to activate adenylate cyclase as well as by receptor loss.(ABSTRACT TRUNCATED AT 250 WORDS)

Presynaptic alpha 2- and postsynaptic beta-adrenoceptor sensitivity in slices of rat neocortex after chronic treatment with various antidepressant drugs

The effect of chronic (4 weeks) treatment of rats with the antidepressant drugs desipramine, maprotiline, chlorimipramine, zimelidine or iprindol on the sensitivity of presynaptic alpha 2- and postsynaptic beta-adrenoceptors in neocortical slices was investigated. Acute (1 day) treatment with the antidepressants did not affect the efflux of cyclic-AMP induced by isoprenaline (1 microM) from neocortical slices, while after chronic treatment the efflux of cyclic AMP was consistently reduced. Following acute administration the electrically-evoked release of [3H]noradrenaline (NA) from radiolabelled cortical slices remained unchanged. Upon chronic treatment with desipramine the release of [3H]NA was enhanced by about 45%. In contrast, after chronic treatment with maprotiline or chlorimipramine the electrically-evoked release of [3H]NA was not affected, whereas release was even slightly reduced after chronic administration of zimelidine or iprindol. In all cases, however, a similar inhibitory effect of exogenous NA (0.1 microM) on the release of [3H]NA was found. These data indicate that the desensitization of postsynaptic beta-adrenoceptors in rat brain after chronic treatment with antidepressant drugs is not paralleled by a reduction of presynaptic alpha 2-adrenoceptor sensitivity.

Involvement of catecholamines in haemophilus influenzae induced decrease of ?-adrenoceptor function

The deeper airways of patients with asthmatic bronchitis are often infected with Haemophilus influenzae. Vaccination of guinea pigs with H. influenzae resulted in a significant impairment of the isoproterenol induced relaxation of isolated tracheal spirals by approximately 50% 4 days following vaccination. In the present study we further investigated the effects of some drugs affecting catecholamine release on the H. influenzae induced functional desensitization of tracheal spirals. Benserazide, an inhibitor of dopa-decarboxylase, completely prevented the reduction in isoproterenol-induced relaxation after H. influenzae vaccination, while no effect on relaxation of tracheal spirals from control animals was detected. On the other hand, inhibiting the re-uptake of catecholamines with desipramine did not influence the relaxation in the H. influenzae vaccinated tracheal spirals. Treatment of control animals with desipramine however resulted in a decreased relaxation of the isolated spirals by 40%. One day following vaccination with H. influenzae the level of norepinephrine in lung tissue was significantly elevated by 71%, and in plasma by 77%, while after 4 days no significant effects were observed. The spontaneous release of norepinephrine, epinephrine and dopamine of tracheal incubates was increased at days 1 and 4 following vaccination. The release of catecholamines from minced lung incubates of H. influenzae pretreated guinea pigs did not differ from that of controls. On the basis of these results it may be suggested that catecholamine metabolism is changed in lungs from H. influenzae vaccinated animals. Catecholamines, accordingly may play a role in the desensitization of beta-adrenoceptors by H. influenzae.

Mepacrine treatment prevents immobilization-induced desensitization of beta-adrenergic receptors in rat hypothalamus and brain stem

Forced immobilization is a severe stress in rats which diminishes levels of epinephrine in specific nuclei in the hypothalamus and brain stem, suggesting that release of epinephrine is stimulated to a rate which exceeds the rate of its replacement. In the pineal gland, frog erythrocytes, C 6 astrocytoma cells and rat brain, β-adrenoceptor agonists appear to regulate the number of their receptors. Exposure to high concentrations of an agonist leads to apparent decrease in receptors reflected by a decrease in maximal specific binding of antagonists. The apparent decreases in receptors have been shown to be attended by decreases in physiologic responsiveness. In C 6 astrocytoma cells, β-agonists stimulate methylation of phosphatidylethanolamine to increase formation of membrane phosphatidylcholine which in turn appears to enhance activation of adenyl cyclase. Interference with the metabolism of phospholipids by exposure to phospholipase A 2 inhibitor, mepacrine (quinacrine), prevents agonist-induced desensitization of β-adrenoceptors in astrocytoma cells. In the present study repeated immobilization stress has been found to decrease significantly the number of β-adrenoceptors in hypothalamus and brain stem while increasing the number ofα 2-adrenoceptors. The desensitization of beta-adrenoceptors was prevented by treatment with mepacrine.