Changes In Beta-adrenergic Receptors Research Articles

Evidence for Coexistence of Three β-Adrenoceptor Subtypes in Human Peripheral Lymphocytes

Peripheral circulating lymphocytes are easily accessible cells for investigating changes in beta-adrenergic receptors (ADRBs) in humans, but previous reports indicate that these cells only express ADRB2. This study aimed to investigate whether ADRB1 and ADRB3 were expressed in peripheral lymphocytes and the changes of ADRBs in congestive heart failure. Our study demonstrates that ADRB1, ADRB2, and ADRB3 coexist in human peripheral lymphocytes, with differential binding property and expression level. Patients with congestive heart failure had significantly decreased total ADRB density and mRNA levels of ADRB1 and ADRB2 genes, but not ADRB3, compared with healthy subjects. The levels of mRNA of ADRB1 and ADRB2 in peripheral lymphocytes from patients with congestive heart disease were significantly increased after drug treatment. Our study, for the first time, indicates that human peripheral lymphocytes coexpress ADRB1, ADRB2, and ADRB3, which has important implications for precisely predicting clinical response to drug therapy in congestive heart failure.

Enhanced β-adrenergic receptors in the brain and pancreas during pancreatic regeneration in weanling rats

Adrenergic stimulation has an important role in the pancreatic beta-cell proliferation and insulin secretion. In the present study, we have investigated how sympathetic system regulates the pancreatic regeneration by analyzing Epinephrine (EPI), Norepinephrine (NE) and beta-adrenergic receptor changes in the brain as well as in the pancreas. EPI and NE showed a significant decrease in the brain regions, pancreas and plasma at 72 hrs after partial pancreatectomy. We observed an increase in the circulating insulin levels at 72 hrs. Scatchard analysis using [(3)H] propranolol showed a significant increase in the number of both the low affinity and high affinity beta-adrenergic receptors in cerebral cortex and hypothalamus of partially pancreatectomised rats during peak DNA synthesis. The affinity of the receptors decreased significantly in the low and high affinity receptors of cerebral cortex and the high affinity hypothalamic receptors. In the brain stem, low affinity receptors were increased significantly during regeneration whereas there was no change in the high affinity receptors. The pancreatic beta-adrenergic receptors were also up regulated at 72 hrs after partial pancreatectomy. In vitro studies showed that beta-adrenergic receptors are positive regulators of islet cell proliferation and insulin secretion. Thus our results suggest that the beta-adrenergic receptors are functionally enhanced during pancreatic regeneration, which in turn increases pancreatic beta-cell proliferation and insulin secretion in weanling rats.

Alterations in adrenergic receptor signaling in heart failure.

In the failing heart, several changes occur in cardiac adrenergic receptor-signal transduction pathways. The most striking of these changes occur in beta-ARs, and of the changes in beta-adrenergic receptors, beta1-receptor down-regulation is the most prominent. Other changes include uncoupling of beta2-adrenergic receptors and increased activity of the inhibitory G-protein, Gi. Most of these changes appear to be related to increased activity of the adrenergic nervous system, i.e. increased exposure to norepinephrine. Antagonists of the adrenergic nervous system improve left ventricular function and outcome in patients with heart failure. This fact supports the notion that activation of these neurohormonal systems exerts a net long-term detrimental effect on the natural history of chronic heart failure and that myocardial adrenergic desensitization phenomena are at least partially adaptive in the setting of left ventricular dysfunction.

14 Changes in beta-adrenergic receptors during left ventricular hypertrophy caused by chronic pressure or volume overload

14 Changes in beta-adrenergic receptors during left ventricular hypertrophy caused by chronic pressure or volume overload

Cardiac adenylyl cyclase, beta-adrenergic receptors, and G proteins in salt-sensitive hypertension.

The present study investigated whether high salt intake (8%) in Dahl salt-sensitive and salt-resistant rats with and without hypertension produces a heterologous desensitization of cardiac adenylyl cyclase as observed in various types of hypertension and human heart failure. In membranes from Dahl salt-sensitive rats on a high-salt diet (8%) basal, isoproterenol-, 5'-guanylylimidodiphosphate-, and forskolin-stimulated adenylyl cyclase was reduced compared with the low-salt (0.4%) group and Dahl salt-resistant rats on either 0.4% or 8% sodium chloride. The activity of the catalyst was depressed, and the expression of the immunodetectable inhibitory G proteins Gi alpha was increased in Dahl salt-sensitive rats on 8% sodium chloride, whereas the density of beta-adrenergic receptors and the activity of the stimulatory G protein Gs alpha reconstituted into Gs alpha-deficient S49 cyc- mouse lymphoma cell membranes were unchanged in any condition studied. We conclude that high salt intake in salt-sensitive hypertensive Dahl rats produces hypertension, cardiac hypertrophy, and heterologous desensitization of cardiac adenylyl cyclase. The latter alteration is due to an increase of Gi alpha proteins and a depressed catalyst activity of adenylyl cyclase. The results demonstrate that heterologous adenylyl cyclase desensitization can precede the development of contractile dysfunction in later stages and can occur independently of changes in beta-adrenergic receptors.

Changes in Beta-Adrenergic Receptors under Long-Term Application of Ritodrine in Pregnant-Rat Myometrium

Changes in beta-adrenergic receptor density under long-term application of ritodrine were investigated using longitudinal muscle strips of day-21-pregnant-rat myometrium. Membrane (-)[3H]dihydroalprenolol binding sites were increased by 3.1 x 10(-8) M ritodrine, with a maximum increase at 3.1 x 10(-7) M, followed by a gradual decrease at higher concentrations. Under long-term application of a lower dose (1.5 x 10(-7) M) of ritodrine, the number of the binding sites increased up to 20 min, then decreased to the control level and fluctuated after 90 min. These results suggest that at lower doses, ritodrine might activate adenylate cyclase, resulting in conformational changes in the receptors, and that receptor density might vary in parallel with cellular cAMP production under long-term application of ritodrine.

Effect of forced-running stress on beta-adrenergic receptors in rat brain regions and liver.

Rats were exposed to forced-running stress for 1 day, 3 days or a long term (approximately 2 weeks), and beta-adrenergic receptor binding was then assayed using [3H]dihydroalprenolol (DHA) in six brain regions and the liver. In the pons + med.obl., hypothalamus and midbrain, a reduction in beta-adrenergic receptor density was first evident on day 1. In contrast, a decrease in beta-adrenergic receptor density in the cerebral cortex and hippocampus was first evident on day 3. Decreased [3H]DHA binding in the pons + med.obl., cerebral cortex and hippocampus subsequently plateaued for the duration of the forced-running stress. In the midbrain and hypothalamus, however, decreased [3H]DHA binding subsequently returned to control levels despite the exposure to the forced-running stress. In the cerebellum and the liver, [3H]DHA binding did not change significantly throughout the stress. These results indicate that the forced-running stress induces both the time- and region-specific changes in beta-adrenergic receptors. Moreover, the rats showed either a behavioral depression or a spontaneous recovery of running activity during the 2 weeks following the end of the long-term stress. Thus, we also examined the relationship of beta-adrenergic receptors to these behavioral differences. [3H]DHA binding for the behavioral depression group was lower in the hippocampus and higher in the liver than for the spontaneous recovery group.

Arterial Oxygen Consumption After Hemorrhagic Shock: The Effect of β-Adrenergic Agonist

Oxygen consumption of the rabbit femoral artery after hemorrhagic shock was studied. Hemorrhagic shock was initiated and maintained at 60 mm Hg of systolic blood pressure for 2 hours. A significant reduction in femoral artery oxygen consumption was observed after hemorrhagic shock (1.64 +/- 0.14 microliter/g/hr) when compared to oxygen consumption in the normal condition (2.52 +/- 0.22 microliter/g/hr). Application of the beta-agonist isoproterenol significantly increased oxygen consumption in the isolated femoral artery after hemorrhagic shock (2.66 +/- 0.20 microliter/g/hr), but did not exceed the normal values recorded without stimulation. Also, isoproterenol significantly increased oxygen consumption in the femoral artery of nonhemorrhagic condition (4.84 +/- 0.42 microliter/g/hr). The increase in oxygen consumption conditioned by isoproterenol was significantly lower after hemorrhagic shock compared with values in the nonhemorrhagic state. The data suggest that oxygen consumption is regulated by beta-adrenergic receptors, and the phenomenon of diminished oxygen consumption after hemorrhagic shock probably occurs because of changes in beta-adrenergic receptors, causing the appearance of change in the mechanism of oxygen consumption.

Differential effects of chronic antidepressants in behavioural tests of β-adrenergic and GABAB receptor function

The effects of chronic administration of anti-depressant drugs (28 days SC via Alzet 2ML4 osmotic minipumps) on the functional sensitivity of beta-adrenergic and GABA receptors have been assessed. Phenelzine (10 mg/kg), tranylcypromine (1 mg/kg), imipramine (30 mg/kg) and desmethylimipramine (10 mg/kg) attenuated the motor-suppressant effects of salbutamol (3 mg/kg) observed at 21-22 days of drug administration. No changes in the motor-suppressant effects of the GABA prodrug progabide (50 mg/kg) or the GABAB agonist (+/-)-baclofen (5 mg/kg) were induced by these antidepressants. These findings extend and confirm previous reports of functional changes in beta-adrenergic receptors but not of GABAB receptors following chronic antidepressant treatment.

Biventricular down-regulation of beta-adrenergic receptors in right ventricular hypertrophy induced by monocrotaline.

To investigate the changes in beta-adrenergic receptors in the right and left ventricles during the development of cardiac hypertrophy, right ventricular hypertrophy (RVH) was induced in rats by a single injection of monocrotaline (M) (60 mg/kg). The rats demonstrated RVH at 2 weeks and severe RVH with congestive heart failure at 4 weeks after the injection. Changes in the beta-adrenergic receptors in the membrane fractions of both ventricles were examined in the M-treated vs. control rats using 125I-iodocyanopindolol (ICYP). Plasma and tissue catecholamines were measured. Plasma levels of norepinephrine (NE) and epinephrine (EP) showed no change at 2 weeks after the injection. However, at 4 weeks there was a significant increase in NE (p less than 0.01), but not in EP. There was no significant change in NE or EP in either ventricle at 2 weeks vs. the controls. However, at 4 weeks there was a significant increase in both NE and EP in the right ventricle (RV) (p less than 0.05). NE in the left ventricle (LV) tended to increase, but not a significant extent (p less than 0.1). With the development of RVH at 2 weeks, the maximum number of binding sites (Bmax) in both the RV and LV showed a significant decrease (p less than 0.01). There was a 21% decrease of Bmax in the RV and 15% in the LV at 2 weeks vs. the controls. At 4 weeks, the changes in Bmax were more prominent, with a 57% decrease in the RV and 22% in the LV.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in beta-adrenergic receptors and catecholamlnes in the heart of rats submitted to various types of trauma

Changes in beta-adrenergic receptors and catecholamlnes in the heart of rats submitted to various types of trauma

Viral Pneumonia Attenuates Adenylate Cyclase But Not Beta-Adrenergic Receptors in Murine Lung

Respiratory infections provoke increased airway reactivity in both asthmatic and otherwise healthy subjects in part through impaired beta-adrenergic relaxation of bronchial and tracheal muscle. The precise mechanism remains obscure, but some studies report a decrease in the number of beta-adrenergic receptors. The present study was designed to assess the effect of viral respiratory infection with influenza A on lung beta-adrenergic receptors and adenylate cyclase activation in mice under two separate protocols. First, to determine whether changes are due to a local or systemic effect, we compared mice with influenza infections limited to the upper respiratory tract to mice with infection of the total respiratory tract. Four days after upper respiratory tract infection there were no changes in either isoproterenol- or NaF-stimulated adenylate cyclase activity. In contrast, there was an 82% decrease in isoproterenol- and a 25% decrease in NaF-stimulated adenylate cyclase activity on the fourth day after total respiratory tract infection. There were no changes in beta-adrenergic receptors or receptor coupling to adenylate cyclase with either type of infection. Our second protocol compared acutely infected mice to postrecovery mice. Twelve days after infection the virus was no longer present in the lungs, and adenylate cyclase activity was restored to normal. These data suggest that viral respiratory infection may impair airway function through attenuation of receptor and postreceptor activation of adenylate cyclase activity.

Human Chorionic Gonadotropin-Induced Heterologous Desensitization of Rabbit Luteal Adenylyl Cyclase Is Associated with Altered Receptor and G-Protein Function*

We injected hCG into pseudopregnant rabbits on day 7 of pseudopregnancy and analyzed changes in the components of luteal adenylyl cyclase system in order to determine which components are responsible for altered hormonal responsiveness upon desensitization. hCG-induced desensitization was homologous (loss of responsiveness to LH) early (first 6 h), then became heterologous (partial loss of responsiveness to catecholamines) later (12-48 h). The total number of LH receptors was reduced approximately 30% 3 h after treatment at a time when LH stimulation of adenylyl cyclase activity was not altered. Total LH receptors remained at this level until 24 h, when total receptors were reduced by 88%. While total LH receptor number remained constant, LH-stimulated adenylyl cyclase activity was declining to 57% of the control value at 12 h. Available unoccupied LH receptors were reduced by 96% at 12 h. The affinity of the occupied receptors was reduced 4-fold before down-regulation. The changes in beta-adrenergic receptor number paralleled the changes in catecholamine responsiveness. hCG treatment also altered luteal G-protein function, as assessed by reconstitution of adenylyl cyclase activity in S49 cyc- lymphoma membranes and ADP ribosylation by cholera and pertussis toxins. Isoproterenol (ISO)-reconstituting activities of luteal Gs (the stimulatory G-protein of adenylyl cyclase) were depressed by 65% 12-48 h after hCG treatment, the same time as reduced catecholamine responsiveness. In contrast, NaF-reconstituting activities were at control levels at 12 h and reduced by 55% at 24 and 48 h. Pertussis toxin's ability to ADP ribosylate alpha i 40 was increased 3 and 6 h after treatment, while cholera toxin's ability to ADP ribosylate alpha s 45 was reduced throughout the study period. These studies demonstrate that hCG-induced heterologous desensitization results in a complex series of changes in beta-adrenergic and LH receptors as well as G-protein function, which account for the altered hormonal responsiveness.

Long-term effects of basal forebrain lesions on cholinergic, noradrenergic and serotonergic markers in mouse neocortex

Lesions of basal forebrain cholinergic neurons projecting to cerebral cortex and hippocampus have recently been exploited as animal models for some of the neurochemical and behavioral deficits of Alzheimer's disease. We have observed that electrolytic lesions of cholinergic basal forebrain nuclei can lead to morphological plasticity in adult mouse cortex. In the present study, the acute and chronic sequelae of basal forebrain electrolytic lesion on cortical synaptic chemistry have been examined. In addition to choline acetyltransferase (ChAT) activity, levels of norepinephrine and of serotonin were reduced within a week after the lesion. Recovery of ChAT activity and of serotonin levels began within a month after the lesion. Serotonin type 2 receptor binding exhibited an acute reduction after the lesion in ipsilateral cortex, followed later by a chronic bilateral decrease. No significant changes in beta-adrenergic receptors were apparent at any time after the lesion despite a permanent and bilateral reduction of norepinephrine levels after the lesion. The potential significance of these results for cortical plasticity regulation and Alzheimer's disease is discussed.

Modulation of the β-Adrenergic Receptor-Coupled Adenylate Cyclase by Chemical Inducers of Differentiation: Effects on β Receptors and the Inhibitory Regulatory Protein Gi

Several drugs known to induce differentiation in tumor cells were analyzed for their effects on the beta-adrenergic receptor-coupled adenylate cyclase system in two human carcinoma cell lines, HeLa and A431. Each of the drugs was tested alone or in combination with sodium butyrate (NaBu), a known inducer of this signal transduction system. Puromycine amino nucleoside (PMAN) caused the largest increase in beta-adrenergic receptors in HeLa cells followed by hexamethylenebisacetamide (HMBA) whereas 5'-azacytidine (5AZC) was ineffective. In addition, PMAN but not the others acted together with NaBu to elevate receptor levels 12-fold over control values. In contrast, HMBA and 5AZC were much more effective on A431 cells, PMAN caused only a slight increase in beta receptors and none of the drugs acted in concert with NaBu. The increase in beta receptors was usually accompanied by a corresponding increase in isoproterenol-stimulated adenylate cyclase activity. These effects of the drugs appeared to require protein synthesis as they were blocked by cycloheximide. In addition, some of the drugs caused a substantial decrease in basal adenylate cyclase activity. This effect on basal activity was abolished in cells treated with pertussis toxin, which ADP-ribosylates the inhibitory GTP-binding protein, Gi. Both HeLa and A431 cells contained a 41 kDalton substrate for the toxin which corresponds to the alpha subunit of Gi. The Gi subunit was ADP-ribosylated by the toxin to a similar extent in membranes from control and drug-treated cells. Thus, the drugs appear to induce quantitative changes in beta-adrenergic receptors and qualitative changes in Gi which results in a highly responsive beta-adrenergic-stimulated adenylate cyclase.

Alpha- and beta-adrenergic receptors in proximal tubules of rat kidney.

Proximal tubules were isolated from the rat kidney by collagenase digestion of the cortical tissue followed by Percoll gradient centrifugation. Microscopic and hormone-stimulated adenylate cyclase activity studies proved the purity of the preparation. [3H]Prazosin, [3H]rauwolscine, and [125I]iodocyanopindolol were used to identify and quantitate respectively the alpha 1-, alpha 2- and beta-adrenergic receptors. Proximal tubular (F4) particulate fraction was compared against other cortical nephron segment (F1, F2) fractions and the total collagenase-digested cortex particulate suspension (Ft). Proximal tubules were enriched in alpha 1- and alpha 2-adrenergic receptors compared with Ft (alpha 1-receptor, 100.4 +/- 4.5 vs. 87.4 +/- 4.9; alpha 2-receptor, 250 +/- 16.2 vs. 185.1 +/- 12 fmol/mg protein). The fractions enriched in glomeruli and distal tubular segments (F1, F2) had relatively low concentrations of alpha 1- and alpha 2-adrenergic receptors. In contrast, beta-adrenergic receptor concentration in the proximal tubules was approximately 25% of that in the Ft fraction and approximately 10% of that in the F1 fraction. Isoproterenol-stimulated adenylate cyclase activities in the different fractions corroborated well with the pattern suggested by the [125I]iodocyanopindolol binding studies. Our results suggest that whole-cortex preparation radioligand binding studies may reflect proximal tubular alpha 1- and alpha 2-adrenergic receptor changes quite well. They may, however, miss or give erroneous impressions about beta-adrenergic receptor changes occurring in different cortical nephron segments.

Beta-adrenergic receptor changes during tertatolol treatment in healthy volunteers: relevance for beta-blocking therapy.

Tertatolol is a new potent beta-blocker without intrinsic sympathomimetic activity and lacking beta 1/beta 2 receptor subtype selectivity. Tertatolol was found in vitro to be a competitive inhibitor of beta-adrenergic receptors in competitive binding experiments. However, the density of beta-adrenergic receptors on intact human lymphocytes preincubated with tertatolol was reduced. When given at therapeutic doses (5 mg/day) to human volunteers, it induced a reduction in the number of beta-adrenergic receptors (Bmax) without any change in the affinity (KD) for intact lymphocytes (beta-adrenergic receptors were measured by the specific binding of 3H-CGP 12177). This effect was seen 7 h (-54%), 24 h (-35%) and 48 h (-30%) after a single drug dose. A similar receptor reduction was observed 7 h (-42%), 24 h (-37%) and 48 h (-15%) after 14 daily doses of the drug. In parallel, the pharmacological efficacy of the drug was evident from the reduction in heart rate in supine and upright positions and after submaximal exercise; heart rate was reduced to the same extent after single or repeated drug doses. The reduction in receptor number correlated with the reduction in heart rate in both the supine (p less than 0.001) and upright (p less than 0.01) positions and after exercise (p less than 0.02). We conclude that tertatolol, besides competitively inhibiting beta-adrenergic receptors, induces a marked and lasting decrease in beta-adrenergic receptor number. This effect may be important for the beta-blocking effects of this drug.

Characterization of a potentially reversible increase in beta-adrenergic receptors in isolated, neonatal rat cardiac myocytes with impaired energy metabolism.

Previous studies have reported that the numbers of beta- and alpha-adrenergic receptors increase in ischemic myocardium. In vivo studies have raised questions regarding the mechanisms involved in the adrenergic receptor alterations and the consequences of these alterations. The purpose of this study was to evaluate potential relationships among beta-adrenergic receptor changes, high energy phosphate reduction, and severity of cell injury in cultured neonatal rat myocytes treated with metabolic inhibitors. The potential for reversal of the receptor changes also was addressed. Binding parameters were measured using [125I]iodocyanopindolol. After 4 hours incubation in potassium cyanide and 2-deoxyglucose, there was a 43% increase in beta-adrenergic receptor number, 41% decrease in adenosine triphosphate, and minimal morphological change in myocytes. Twenty-four hours after removal of the inhibitors, myocytes exhibited a return to normal of the receptor number and adenosine triphosphate level. Iodoacetate treatment for up to 3 hours resulted in marked reduction in adenosine triphosphate and increasing severity of cell injury. The number of beta-adrenergic receptors was unchanged at 1.2 hours, increased at 1.5-2 hours, and decreased at 3 hours. Thus: beta-adrenergic receptor density increases during relatively early stages of injury in metabolically impaired myocytes with reduced adenosine triphosphate levels and decreases subsequently, after the myocytes become irreversibly injured; the increased beta-adrenergic receptor density in moderately injured myocytes can be reversed upon removal of the injurious agent and restoration of the cellular adenosine triphosphate level; and changes in catecholamines mediated by an intact nervous system are not required for an increase in beta-adrenergic receptor density in the setting of impaired energy metabolism.

Cardiac beta-adrenergic receptors in salt-dependent genetic hypertension.

The pattern of cardiac beta-adrenergic receptor changes in different hypertrophy models varies according to the pathophysiology. In salt-sensitive Dahl rats, high dietary salt intake leads to a moderate degree of cardiac hypertrophy associated with increased numbers of cardiac beta-adrenergic receptors but unchanged affinity for agonists. Isoproterenol-stimulated cardiac adenylate cyclase is also higher in salt-loaded hypertensive rats without any change in basal or NaF-stimulated activities. In contrast, neither beta-adrenergic receptors nor adenylate cyclase activities are affected by variations in dietary salt in salt-resistant Dahl rats. The extent of isoproterenol-induced down regulation of beta-adrenergic receptors on isolated cardiac myocytes as well as the recovery from this down regulation is not significantly different in either strain of Dahl rats and is not influenced by dietary salt. The enhancement of beta-adrenergic pathways in salt-dependent genetic hypertension may be involved both in the initiation of cardiac hypertrophy and the preservation of contractile function.

Agonist-induced changes in beta-adrenergic receptors on intact cells.

Competition by beta-adrenergic agonists and antagonists for 125I-pindolol binding sites on intact cells (1321N1 human astrocytoma and C62B rat glioma) was measured using short time binding assays as previously described (Toews, M. L., Harden, T. K., and Perkins, J. P. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 3553-3557). Preincubation of cells with agonists converted about half of the cellular beta-adrenergic receptors from a form exhibiting high affinity for the agonists isoproterenol and epinephrine and the antagonist sotalol to a form exhibiting much lower apparent affinity for these ligands in short time assays. Exposure to agonists did not alter the affinity of receptors for the antagonist metoprolol. This change in the ligand binding properties of the receptor was rapid (t1/2 = 1-2 min following a lag of about 0.5 min), reversible (t1/2 = 6-8 min), and dependent on the agonist concentration present during the preincubation (K0.5 = 15 nM for isoproterenol). Both isoproterenol and sotalol attained equilibrium with the high affinity receptors very rapidly but equilibrated only slowly with those receptors exhibiting low apparent affinity in short time assays. These results are interpreted in terms of a model which postulates that both the low apparent affinity in short time assays and the subsequent slow equilibration of hydrophilic ligands with these receptors result from agonist-induced internalization of a fraction of cell surface beta-adrenergic receptors. The relationship of this change in receptor binding properties to other aspects of agonist-induced desensitization of the beta-adrenergic receptor-coupled adenylate cyclase system is discussed.