Beta-adrenoceptor Binding Sites Research Articles

Characterization of beta-adrenergic receptors in duck cerebral cortex

Beta-adrenoceptor binding sites were characterized in duck cerebral cortex by an in vitro binding technique, using 3H-dihydroalprenolol (3HDHA) as a receptor-specific radioligand. The specific binding of 3HDHA to duck cerebral cortical membranes was found to be rapid, stable, saturable, reversible, and of high affinity. Saturation analysis resulted in a linear Scatchard plot suggesting binding to a single class of receptor binding sites with high affinity (Kd = 1.18 nM) and high capacity (Bmax = 162 fmol/mg protein). Competition studies showed the following relative rank order of potency of various compounds to inhibit the 3HDHA binding: antagonists ­ICI 118,551 > S(-)-propranolol >> betaxolol, yohimbine, WB-4101, prazosin, mianserine; agonists - isoprenaline approximately equal to fenoterol> salbutamol >> clonidine, phenylephrine. The obtained data suggest that in duck cerebral cortex beta-adrenergic receptors (like those described in brains of chick and pigeon) are of the beta2 subtype. This is in contrast to what has been reported for the mammalian brain, where - among beta-adrenoceptors - the beta1 subtype is predominant.

Dynamic regulation of beta-adrenergic receptors by endothelin-1 in smooth-muscle cells.

Elevated endothelin-1 (ET-1) levels are found in atherosclerosis, myocardial ischemia, and heart failure, and are correlated with increased mortality rates. Contrary to expectations, elevations of endogenous ET-1 levels in transgenic mice are not associated with increases in arterial blood pressure or with vasospasm, although these effects can be observed after i.v. ET-1 administration. The aim of this study was to determine the regulatory effects of ET-1 on the expression of vasodilator beta-adrenergic receptors and their ability to activate adenylyl cyclase. Smooth-muscle cells were incubated with ET-1 (10(-7) mol/L) for 3 days. The density of ET-1 or beta-adrenergic receptor binding sites was determined using a radioligand binding procedure. Adenylyl cyclase activity was measured to assess any functional changes in the beta-adrenergic receptor density. ET-1 incubation reduced ET-1 binding sites by 70%. In contrast, the beta-adrenergic receptor density increased from 354 +/- 35 to 538 +/- 50 fmol/mg protein (p < 0.01; n = 7) after 3 days. ET-1 increased beta-adrenergic receptors dose-dependently. Incubation with ET-1 for different periods of time showed an initial decrease of 30% after 6 h of ET-1 incubation. However, after 24 h ET-1 induced an increase of beta-adrenergic receptors, reaching a maximal amount after 48 h. An increased stimulation of beta-adrenergic receptor-activated adenylyl cyclase was observed after 3-day ET-1 incubation compared to controls. These data demonstrate that chronic ET receptor activation by ET-1 results in a functionally significant increase in beta-adrenergic receptor density and adenylyl cyclase activity.

Evidence for direct interaction of ketamine with alpha 1- and beta 2-adrenoceptors.

1. Ketamine has a number of effects that suggest that it may interact with alpha- and beta-adrenoceptors. To date, the experimental evidence for this has been indirect and has been based on physiological studies using competitive blocking agents. In the present study we sought to determine from receptor binding studies whether ketamine binds directly to alpha- and beta-adrenoceptors. 2. Membrane preparations of alpha 1- and beta 2-adrenergic binding sites were obtained from urinary bladder and urethrae of sheep. These binding sites were characterized by saturation analyses using [3H]-prazosin for alpha 1-adrenoceptor binding sites and [125I]-cyanopindolol (CYP) for the beta 2-adrenoceptor binding sites. The receptors were further characterized by displacement studies using selective and non-selective antagonists. 3. Studies in which ketamine was used to displace [3H]-prazosin revealed a Kd of 3.40 +/- 1.23 x 10(-3) mol/L for ketamine binding to alpha 1-adrenoceptors. Displacement studies of [125I]-CYP by ketamine showed a Kd of 0.35 +/- 0.03 x 10(-3) mol/L for ketamine binding to beta 2-adrenoceptors. 4. We conclude that ketamine interacts directly with both alpha 1- and beta 2-adrenoceptors and that such interactions probably explain the reported effects of this agent on the vasculature and the bronchial tree.

Effects of beta 2-agonist- and dexamethasone-treatment on relaxation and regulation of beta-adrenoceptors in human bronchi and lung tissue.

1. Long-term treatment with beta 2-adrenoceptor agonists can lead to a decreased therapeutic efficacy of bronchodilatation in patients with obstructive pulmonary disease. In order to examine whether or not this is due to beta-adrenoceptor desensitization, human bronchial muscle relaxation was studied in isolated bronchial rings after pretreatment with beta 2-adrenoceptor agonists. Additionally, the influence of pretreatment with dexamethasone on desensitization was studied. 2. The effect of beta 2-agonist incubation alone and after coincubation with dexamethasone on density and affinity of beta-adrenoceptors was investigated by radioligand binding experiments. 3. In human isolated bronchi, isoprenaline induces a time- and concentration-dependent beta-adrenoceptor desensitization as judged from maximal reduction in potency by a factor of 7 and reduction of 73 +/- 4% in efficacy of isoprenaline to relax human bronchial smooth muscle. 4. After an incubation period of 60 min with 100 mumol l-1 terbutaline, a significant decline in its relaxing efficacy (81 +/- 8%) and potency (by a factor 5.5) occurred. 5. Incubation with 30 mumol l-1 isoprenaline for 60 min did not impair the maximal effect of a subsequent aminophylline response but led to an increase in potency (factor 4.4). 6. Coincubation of dexamethasone with isoprenaline (120 min; 30 mumol l-1) preserved the effect of isoprenaline on relaxation (129 +/- 15%). 7. In radioligand binding experiments, pretreatment of lung tissue for 60 min with isoprenaline (30 mumol l-1) resulted in a decrease in beta-adrenoceptor binding sites (Bmax) to 64 +/- 1.6% (P < 0.05), while the antagonist affinity (KD) for [3H]-CGP-12177 remained unchanged. 8. In contrast, radioligand binding studies on lung tissue pretreated with either dexamethasone (30 mumol l-1) or isoprenaline (30 mumol l-1) plus dexamethasone (30 mumol l-1) for 120 min did not lead to a significant change of Bmax (160 +/- 22.1% vs 142.3 +/- 28.7%) or KD (5.0 nmol l-1 vs 3.5 nmol l-1) compared to the controls. 9. In conclusion, pretreatment of human bronchi with beta-adrenoceptor agonists leads to functional desensitization and, in lung tissue, to down-regulation of beta-adrenoceptors. This effect can be counteracted by additional administration of dexamethasone. Our model of desensitization has proved useful for the identification of mechanisms of beta-adrenoceptor desensitization and could be relevant for the evaluation of therapeutic strategies to counteract undesirable effects of long-term beta-adrenoceptor stimulation.

Androgen support of lacrimal gland function.

The effects of dihydrotestosterone (DHT) (1 mg/kg) on biochemical parameters related to lacrimal secretion, basal tear flow rate, and pilocarpine-stimulated lacrimal gland fluid secretion, in mature ovariectomized rabbits were studied. The effects of the synthetic estrogen diethylstilbestrol (DES) (100 micrograms/kg), on lacrimal gland biochemical parameters in normal mature female rabbits was also studied. Ovariectomy decreased the total serum levels of testosterone (T) by 88.5% and androstenedione by 35.9%, without changing the levels of dehydroepiandrosterone (DHEA) of its sulfate. Ovariectomy caused a significant regression of the lacrimal glands, decreasing total DNA by 35%, and total protein by 22%. DHT treatment of ovariectomized animals prevented lacrimal gland regression, increasing total gland DNA (31%) and total protein (18%). DHT treatment also increases Na+, K(+)-ATPase activity (29%) and beta-adrenergic receptor binding sites (23%) compared to the ovariectomized group. DHT increased pilocarpine stimulated lacrimal gland fluid secretion (13.26 +/- 1.47 microL/min) compared to the ovariectomized group (7.72 +/- 0.41 microL/min), but DHT treatment paradoxically decreased basal tear flow rate (1.02 +/- 0.04 microL/min) as compared to the ovariectomized rabbits (1.96 +/- 0.12 microL/min). DES decreased the total serum T from 59.33 +/- 10.54 pg/mL to 21.5 +/- 6.06 pg/mL. DES decreased total Na+,K(+)-ATPase by 12% and increased beta-adrenergic receptor binding sites by 83.3%. These results suggest that androgens play a major role in supporting lacrimal gland secretory function. Additionally, they suggest that estrogens may influence certain aspects of lacrimal functions, although it is not clear to what extent those actions are elicited directly or indirectly.

Effect of glucocorticosteroid treatment on glucocorticoid receptor expression in human adipocytes.

The influence of glucocorticoid excess on expression of the glucocorticoid receptor (GR) and beta-adrenoceptor subtype was studied in isolated adipocytes obtained by sc fat biopsies from 17 healthy individuals. The biopsies were taken before and after 7 days of treatment with 25 mg prednisolone, given orally. GR and beta 1- and beta 2-adrenoceptor messenger ribonucleic acid (mRNA) levels were measured with a solution hybridization assay, and the number of beta 1- and beta 2-adrenoceptor binding sites was determined in radioligand binding experiments. GR protein levels were determined by Western blot analysis using an anti-GR antibody. Both GR protein and GR mRNA levels decreased significantly (P < 0.05) by about 50% after treatment, whereas no significant changes were demonstrated in either beta 1- or beta 2-adrenoceptor mRNA levels. The number of beta 2-adrenoceptor-binding sites, however, increased by 70% after treatment (P < 0.05), whereas the number of beta 1-adrenoceptor binding sites was not affected. The affinity of each receptor subtype was not significantly altered by steroid treatment. In conclusion, an in vivo glucocorticoid excess decreases GR mRNA as well as GR protein levels and selectively increases beta 2-adrenoceptor density in sc fat cells of healthy individuals. This indicates that glucocorticoids modulate human adipose metabolism by altering the expression of regulatory proteins at various mRNA and post-mRNA levels.

Effects of weight reduction on the regulation of lipolysis in adipocytes of women with upper-body obesity.

1. The regulation of lipolysis was studied in 14 upper-body obese women aged 26-55 years. Isolated subcutaneous adipocytes from the abdominal region were examined before and after an 8 to 12-week-long weight reduction programme, during which the mean body mass index (kg/m2) of the group was reduced from 36.4 +/- 0.9 to 30.3 +/- 1.0. 2. Fat cell volume was reduced from 891 +/- 39 to 655 +/- 45 pl. The sensitivity to noradrenaline stimulation of lipolysis in vitro increased fivefold after weight reduction. A corresponding increase in sensitivity was found with the beta 2-selective adrenoceptor agonist terbutaline. However, the number of beta 2-adrenoceptor binding sites as assessed by radioligand binding with 125I-labelled cyanopindolol was not changed. No changes were observed when dobutamine (a beta 1-selective adrenoceptor agonist) and clonidine (an alpha 2-adrenoceptor agonist) were used. 3. The basal lipolysis rates decreased by about 50% after weight reduction and the maximum enzyme activity of hormone-sensitive lipase was also reduced by almost 50%. 4. Plasma concentrations of insulin, noradrenaline and total testosterone decreased and sex hormone-binding globulin increased after weight reduction. Calculated apparent free testosterone levels decreased by more than 40% after weight reduction. 5. In conclusion, weight reduction leads to increased efficiency of adipocyte lipolysis with decreased resting lipolysis rate but increased sensitivity to stimulation by catecholamines, which may be attributed to a decreased activity of hormone-sensitive lipase and an increased sensitivity of beta 2-adrenoceptors. Changes in circulating levels of catecholamines, insulin and testosterone may play a role in these modifications of adipocyte function.(ABSTRACT TRUNCATED AT 250 WORDS)

Neuropeptide Y prevents agonist-stimulated increases in contractility.

Neuropeptide Y has been shown to inhibit contractility in the rat heart. Although the reasons for this effect are not known, it is possible that postsynaptic adrenergic mechanisms involving neuropeptide Y may be responsible. To ascertain whether this neuromodulatory effect is possible for decreasing contractility, we investigated the effect of neuropeptide Y on agonist-stimulated contractility of the isolated rat myocardium. Receptor binding studies of purified cardiac membranes showed that incubating membrane in the presence of neuropeptide Y (10(-7) mol/L) decreased the number of alpha-/beta-adrenoceptor binding sites without affecting the affinity of these receptors. Isolated hearts perfused with phenylephrine (10(-5) to 10(-10) mol/L) or isoproterenol (10(-5) to 10(-10) mol/L) in a nonrecirculating Langendorff setup demonstrated a significant increase in contractility over control values, whereas no change in contractility was observed when the hearts were perfused with neuropeptide Y (10(-7) mol/L). However, in the presence of both agonist and neuropeptide Y the increase in contractility previously seen with agonist alone was not evident. Comparisons made with hearts taken from aortic banded rats yielded similar results. Although neuropeptide Y itself was ineffective in decreasing contractility, it prevented the agonists from stimulating contractility when perfused together. We conclude that neuropeptide Y does not directly decrease contractility but prevents agonist-stimulated increases in contractility through alpha-/beta-adrenoceptor pathways. This neuromodulatory effect of neuropeptide Y is unchanged in situations of increased sympathetic activity, such as hypertension.

Effects of chronic infusion of (-)-isoprenaline on rat cardiac muscarinic (M2)-cholinoceptors and beta 1- and beta 2-adrenoceptors.

1. The effects of chronic (-)-isoprenaline infusion (400 micrograms kg-1 h-1; 4 h-14 days) on rat ventricular muscarinic M2-cholinoceptors and beta 1- and beta 2-adrenoceptors was determined. [3H]-N-methylscopolamine (NMS) was used to measure M2-cholinoceptor binding and (-)-[125I]-cyanopindolol (CYP) was used for beta 1- and beta 2-adrenoceptor binding. 2. Chronic infusion of (-)-isoprenaline did not affect either the affinity of [3H]-NMS for M2-cholinoceptors, or the maximal density of these receptors (Bmax) at all treatment periods investigated (4 and 20 h; 7 and 14 days). The affinity of (-)-[125I]-CYP for beta-adrenoceptors was not changed following chronic (-)-isoprenaline infusion (4, 12 and 20 hours; 7 and 14 days). 3. Competition between (-)-[125I]-CYP and the selective beta 1-adrenoceptor antagonist CGP 20712A resulted in biphasic curves at all treatment periods corresponding to the presence of beta 1- and beta 2-adrenoceptor binding sites. beta 1-adrenoceptors made up the greater proportion of beta-adrenoceptors in rat ventricle. There was no change in the ratio of beta 1- to beta 2-adrenoceptors following 4 h (-)-isoprenaline infusion [(beta 1:beta 2), vehicle: 77.5 +/- 4.0%:22.5 +/- 4.0%, n = 6; (-)-isoprenaline: 79.2 +/- 1.2%:20.8 +/- 1.2%, n = 6], however infusion for 12 h increased the ratio [(beta 1:beta 2), vehicle: 65.2 +/- 6.1%:34.8 +/- 6.1%, n = 6; (-)-isoprenaline: 85.6 +/- 1.8%:14.4 +/- 1.8%, n = 6, P < 0.05], which was maintained for longer infusion periods (20 h-14 days). 4. Both beta 1- and beta 2-adrenoceptors were down-regulated in rat ventricular tissue following (-)-isoprenaline infusion. beta 2-adrenoceptors were down-regulated by 12 h (vehicle: 11.8 +/- 2.4 fmol mg protein-1, n = 6; (-)-isoprenaline: 3.5 +/- 0.4 fmol mg protein-1, n = 6, P < 0.05) however beta 1-adrenoceptors were not affected (vehicle: 21.7 +/- 2.0 fmol mg protein-1, n = 6; (-)-isoprenaline: 20.9 +/- 0.6 fmol mg protein-1, n = 6, P > 0.05) until 20 h infusion (vehicle: 19.4 +/- 2.2 fmol mg protein-1, n = 6; (-)-isoprenaline: 12.6 +/- 2.2 fmol mg protein-1, n = 6, P < 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Visualization of beta-adrenoceptor binding sites on human inferior vagal ganglia and their axonal transport along the rat vagus nerve

Because of uncertainties regarding the complete antihypertensive mechanism of action of beta-adrenoceptor antagonists, the present study determined whether vagal afferent neurons of humans and rats possess beta-adrenoceptors. Such a location would provide an appropriate target for beta-blockers to modulate neurotransmission of barosensitive neurons, thereby affecting blood pressure. Therefore, in vitro receptor autoradiography of high-affinity beta-adrenoceptor binding sites was performed on slices of human and rat inferior vagal (nodose) ganglia with [125I]-pindolol. Slide-mounted sections of human and rat inferior vagal ganglia were incubated with [125I]-pindolol in the absence or presence of propranolol (10 mumol/l) to define non-specific binding, atenolol (10 mumol/l) to inhibit binding to beta 1-adrenoceptors, or ICI 118551 (3 nmol/l) to inhibit binding to beta 2-adrenoceptors. Unilateral vagal ligation was also performed in the rat to study whether beta-adrenoceptors are subject to axonal transport along the vagus nerve. [125I]-pindolol bound with > 90% specific binding to sections both of human and of rat inferior vagal ganglia. Specific binding occurred over both neuronal perikarya and nerve fibres. In both species the beta 2-adrenoceptor subtype appeared to predominate, as defined by the differential ability of ICI 118551 (beta 2) and atenolol (beta 1) to inhibit the binding of [125I]-pindolol. Furthermore, unilateral vagal ligation in the rat caused an accumulation of specific binding adjacent to the ligature sites. We conclude that human and rat vagal afferent (and efferent) neurons possess beta-adrenoceptors that potentially could explain the mechanism of action of beta-adrenoceptor antagonists in the therapy of hypertension.

Alterations in beta-adrenoceptors and polyploidy in cultured aortic smooth muscle cells from different age groups of spontaneously hypertensive rats and Wistar-Kyoto rats.

The relationship between the number of beta-adrenoceptors and polyploidy in cultured aortic smooth muscle cells derived from different age groups of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were examined. The number of beta-adrenoceptors, the percentage of multinucleated cells and the incidence of polyploidy from cultured smooth muscle cells derived from SHR and WKY rats aged 3-4, 10-12 and 28-30 weeks were measured. The effect of passaging of the cells on the expression of beta-adrenoceptors and polyploidy on cultured smooth muscle cells from both SHR and WKY rats was also investigated. Receptor binding experiments were carried out using [125]-monoiodocyanopindolol with osmotically lysed cultured aortic smooth muscle cells to investigate the properties of vascular beta-adrenoceptors in SHR and WKY rats. The proportion of polyploid smooth muscle cells was determined by frequency distribution analyses of Feulgen DNA microdensitometric measurements. The incidence of polyploid smooth muscle cells was consistently higher in cells cultured from SHR than in those from WKY rats in all three age groups, with a positive correlation between polyploidy and age in SHR. Furthermore, in all three age groups the number of beta-adrenoceptor binding sites was also higher in cultured smooth muscle cells from SHR than in those from WKY rats. There was no significant difference in the receptor affinity. The increase in beta-adrenoceptor number was associated with an increase in polyploidy, and both of these changes were positively correlated both with the age of the rats from which these cells were derived and with the number of passages. Under cell culture conditions the expression of beta-adrenoceptor density increases with the number of passages in both SHR and WKY rats. Smooth muscle cells derived from older SHR and WKY rats have a greater propensity to develop polyploidy. This trend is significantly accelerated in cultured smooth muscle cells derived from SHR compared with those from WKY rats, suggesting a premature ageing process. These findings suggest that, in cultured smooth muscle cells from SHR and WKY rats, beta-adrenoceptors may influence the expression of polyploidy.

Regulation of beta-adrenergic receptor mRNA in rat C6 glioma cells is sensitive to the state of microtubule assembly.

Microtubule disrupter, colchicine, and microtubule stabilizer, taxol, were used to determine whether microtubules play a role in beta-adrenergic receptor mRNA homeostasis and agonist-induced down-regulation in C6 glioma cells. Colchicine treatment had significant, differential, time-dependent effects on constitutive beta 1- and beta 2-adrenergic receptor mRNA levels. These effects stemmed from the action of colchicine on microtubules, because beta-lumicolchicine, an inactive isomer, had no effect, and nocodazole, a structurally unrelated microtubule disrupter, had similar effects. Colchicine treatment had little effect on the total number of beta-adrenergic receptor binding sites as measured by (-)-[125I]iodopindolol binding, but did alter the relative proportion of beta 1- and beta 2-adrenergic receptor subtypes. Colchicine also had no effect on basal cyclic AMP levels. In contrast to colchicine, taxol treatment had little long-term effect on either beta 1- or beta 2-adrenergic receptor mRNA levels. Taxol antagonized the effects of colchicine on total binding and mRNA levels. Taxol treatment increased basal cyclic AMP levels fourfold and potentiated (-)-isoproterenol-induced cyclic AMP production. Colchicine pretreatment completely inhibited (-)-isoproterenol-induced down-regulation of beta 1-adrenergic receptor mRNA, but not that of beta 2-adrenergic receptor mRNA. Taxol pretreatment had little effect on isoproterenol-induced beta-adrenergic receptor mRNA down-regulation. Colchicine pretreatment also attenuated isoproterenol-induced receptor down-regulation and inhibited agonist-stimulated cyclic AMP production. These effects of colchicine were antagonized by taxol. Whereas the effects of taxol and colchicine on isoproterenol-induced down-regulation of beta-adrenergic receptor mRNA are consistent with their effects on cyclic AMP production, those of colchicine in the absence of stimulation must involve other mechanisms. The data demonstrate that the state of microtubule assembly can affect cyclic AMP levels, beta 1- and beta 2-adrenergic receptor mRNA, and binding site levels in C6 glioma cells.

Response of mouse proximal straight tubule and medullary thick ascending limb to beta-agonist.

Scatchard analysis of (3H)CGP-12177 and (125I)cyanopindolol (CYP) radioligand binding data revealed the presence of specific beta-adrenergic receptor-binding sites on microdissected mouse proximal straight and medullary thick ascending limb (mTAL) tubules. beta-Receptor (10(-6) M isoproterenol) stimulation of isolated perfused mTAL tubules produced a consistent hyperpolarization of the transepithelial potential that was blocked by propranolol (10(-6) M), a beta-receptor antagonist, and furosemide (10(-4) M), an inhibitor of the Na+/K+/2 Cl- triporter. In contrast, there was no electrogenic response to isoproterenol stimulation in isolated perfused proximal straight tubules. In summary, radioligand binding data show that both proximal straight and mTAL tubules possess beta-receptor-binding sites. Electrogenic transport in the mTAL can be modulated by beta-agonists, but there was no detectable electrogenic response to beta-receptor stimulation in proximal straight tubules.

Sequential changes in sympatho-neuronal regulation and contractile function following aortic regurgitation in rabbit heart.

This study was designed to determine the sequential changes in sympatho-neuronal regulation and contractile function of hearts exposed to volume overloading. Aortic regurgitation was produced by perforation of the aortic valve in 20 rabbits. Another 20 rabbits underwent sham operation. They were randomly assigned to a 1-day group, a 1-week group, and a 4-week group. Haemodynamics, myocardial beta-adrenoceptor binding number and catecholamines were measured in each period after production of aortic regurgitation. Left ventricular end-diastolic pressure increased and cardiac output decreased progressively over the week following production of aortic regurgitation, but they returned towards normal during the subsequent 3 weeks. Left ventricular free wall thickness increased 4 weeks after production of aortic regurgitation, and plasma norepinephrine increased 1 day after the procedure. Maximal binding sites of myocardial beta-adrenoceptors (125I-iodocyanopindolol) were decreased 1 day and 1 week after production of aortic regurgitation. Myocardial norepinephrine was also reduced. After 4 weeks, both beta-adrenoceptor binding sites and myocardial norepinephrine content were restored. Alterations in sympatho-neuronal regulation are related to the compensatory processes in volume overloaded hearts.

Effect of glucocorticosteroid treatment on beta-adrenoceptor subtype function in adipocytes from patients with asthma.

1. Adrenoceptor subtype function was studied in isolated adipocytes obtained by subcutaneous fat biopsies from nine patients with mild asthma. The biopsies were taken before and after 7 days treatment with 25 mg of prednisolone given orally. Lipolytic activity after stimulation with various adrenergic agent was measured, using glycerol release as an index of lipolysis. The number of beta 1- and beta 2-adrenoceptor binding sites was determined in radioligand binding experiments and beta 1- and beta 2-adrenoceptor mRNA levels were measured with a solution hybridization assay. 2. Lipolytic sensitivity (ED50) to isoprenaline, a non-selective beta-adrenoceptor agonist, increased 50-fold after treatment (P = 0.04). Sensitivity to terbutaline, a selective beta 2-adrenoceptor agonist, increased 25-fold (P = 0.01), whereas the ED50 values for dobutamine, a selective beta 1-adrenoceptor agonist, did not change significantly. Likewise, the sensitivity to the alpha 2-adrenoceptor agonist, clonidine, and to the drugs acting at post-receptor levels did not change significantly. Basal and maximum lipolytic rates on stimulation were not altered by the treatment. 3. The number of beta 2-adrenoceptor binding sites increased by 60% after treatment (P < 0.05), whereas the beta 1-adrenoceptor binding sites were not affected. The affinity of each receptor subtype for the displacing ligand, ICI 118.551, was not significantly altered by steroids. No significant changes were demonstrated in either beta 1- or beta 2-adrenoceptor mRNA levels. 4. Thus, glucocorticoids selectively increase beta 2-adrenoceptor density and function in patients with asthma, studied by using subcutaneous fat cells as an experimental model.

Evidence for beta-adrenoceptors in rat proximal convoluted tubules.

It is well documented that renal catecholamines, acting as paracrine or autocrine substances, modulate reabsorption of sodium and other substances in renal tubules. However, beta-receptors or the resulting stimulation of adenylate cyclase (AC) have not been measured directly in the proximal convoluted tubules (PCT) of mammalian kidney. In this study, the beta adrenergic receptors in the PCT from rat kidney were characterized pharmacologically by measuring agonist stimulated adenylate cyclase activity. Isoproterenol (ISO) (10 microM) significantly increased AC activity in the microdissected PCT (ISO, 193.0 +/- 23.6 vs basal, 118.7 +/- 14.3 fmol/mm/20; n = 8, p = 0.004). The stimulation of AC in PCT by ISO was blocked by 10 microM propranolol (p < 0.01); it was also blocked by either the beta-1-adrenergic blocker, 10 microM atenolol (p < 0.05), or the beta-2-adrenergic blocker, 10 microM ICI-118551 (p < 0.05). The beta adrenergic receptor binding sites were localized by autoradiography; specific [125I] iodocyanopindolol ([125I] ICYP) binding could be measured in the microdissected rat PCT. In conclusion, these findings provide evidence for beta adrenergic receptors in the PCT in rat kidney by radioligand binding and beta adrenergic agonist stimulated AC activity.

Early changes in second messenger but not receptor binding sites after acute subdural hematoma: an in vitro autoradiographic study.

Neurotransmitter receptor-coupled mechanisms have been recently recognized as important determinants of cell damage after central nervous system (CNS) trauma and ischemia. Many of these receptors exert their intracellular effects via second messenger systems. This study used in vitro autoradiographic radioligand binding to measure beta-adrenergic and muscarinic cholinergic receptors and adenylate cyclase and protein kinase C (PKC) binding sites two h after acute subdural hematoma in rats. Both beta-adrenergic and cholinergic receptor binding sites were unchanged in comparison to controls, while adenylate cyclase binding significantly decreased in the ischemic cortex under the hematoma. These changes may constitute a major limiting factor on receptor-linked therapeutic strategies in trauma and ischemia. Protein kinase C activation significantly increased in the ischemic area under the hematoma in these studies. This appears to be a response to calcium flux, which may be in part glutamate mediated.

"5-HT1R" or 5-HT1D sites? Evidence for 5-HT1D binding sites in rabbit brain.

Radioligand binding studies were performed in membranes of rabbit whole brain and striatum using the novel iodinated radioligand for 5-hydroxytryptamine 5-HT1B and 5-HT1D sites, Serotonin-5-O-Carboxymethyl-Glycyl[125I]Tyrosinamide ([125I]GTI). [125I]GTI labelled a finite number of high affinity sites in rabbit brain membranes, Bmax = 191 +/- 47 fmol/mg protein, pKD (-log mol/l) = 8.50 +/- 0.13, n = 5. The pharmacological profile of [125I]GTI binding was fully comparable to that reported previously in human and other brain preparations known to possess 5-HT1D sites (using either [3H]5-HT or [125I]GTI) and displayed a characteristic rank order of affinity: 5-carboxamido-tryptamine greater than 5-HT = dihydroergotamine greater than or equal to ergotamine greater than or equal to sumatriptan greater than or equal to CGS 12066 greater than or equal to metergoline greater than yohimbine greater than or equal to methysergide greater than ICYP greater than 8-OH-DPAT greater than or equal to CP 93129 greater than (-)pindolol greater than ketanserin greater than isamoltane greater than mesulergine greater than corynanthine greater than buspirone greater than MDL 72222. Autoradiographic studies were performed on rabbit brain slices using [3H]5-HT in the presence of 100 nmol/l 8-OH-DPAT and mesulergine (in order to mask 5-HT1A and 5-HT1C binding sites) and [125I]CYP (iodocyanopindolol) in the presence of 3 mumol/l isoprenaline and 100 nmol/l 8-OH-DPAT (in order to mask beta adrenoceptor and 5-HT1A binding sites). There was no detectable specific binding of [125I]CYP through the brain, thus excluding the presence of 5-HT1B sites in rabbit brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Enhanced myocardial adenylate cyclase activity in spontaneously hypertensive rats.

This study was designed to clarify the state of beta-adrenergic signal transduction and the disordered level of its transduction in hypertensive hearts, using myocardium from spontaneously hypertensive rats (SHR) as a generic model of essential hypertension. Beta-adrenergic receptor binding sites and dissociation constants in the extracted membranes of adult (70-100 days of age) SHR heart were not significantly different from those of Wistar-Kyoto (WKY) rats, the non-hypertensive control. The adenylate cyclase activities stimulated by isoproterenol with GTP, NaF and forskolin were significantly higher in SHR compared to those in WKY. To determine whether differences in signal transduction are natural or are a result of hypertension, we evaluated chronotropic responses in cultured cells of fetal hearts which had not been exposed to hypertension. Fetal cardiac muscle cells of SHR were more sensitive than WKY to isoproterenol stimulation over a wide concentration range. However, there were no statistically significant differences between these two strains with respect to the density of binding sites. These results suggest that in the transduction of adrenergic signals, alterations distal to the beta-receptors are present in the adult hearts of hypertensive rats, and, that the adrenergic signal transduction is already exaggerated in the pre-hypertensive fetal stage.

Brainβ-adrenoceptor binding sites in depressed suicide victims: effects of antidepressant treatment

beta-Adrenoceptor binding sites were measured by saturation binding of [3H]CGP 12177 in nine brain regions from 13 suicides, with a firm retrospective diagnosis of depression, who had been receiving antidepressant drugs, and 11 matched controls. Significantly lower numbers of beta-adrenoceptor binding sites were found in thalamus and temporal cortex (Brodmann area 38), but not in other brain regions, of antidepressant-treated suicides compared to controls. The lower number of beta-adrenoceptor binding sites in thalamus appeared to be related to drug treatment, whereas lower numbers of beta-adrenoceptors in temporal cortex were also found in antidepressant-free suicides.