Beta-adrenergic Receptor Concentration Research Articles

The effects of postnatal anoxia on behaviour and on the muscarinic and beta-adrenergic receptors in the hippocampus of the developing rat

Exposure of rats to 25 min anoxia within 24 h following birth caused behavioural as well as biochemical changes during their development and maturity. Following postnatal anoxia, a significant increase in the concentration of the cholinergic muscarinic receptors in the hippocampus was noted at the early stages of development, between 6 and 20 days of age, but reached normal values at 40 days of age. However, at this age, significant increase in the concentration of beta-adrenergic receptors in the hippocampus was found, which remained significantly high during maturity and adulthood, as compared to controls. Rats submitted postnatally to anoxia exhibited hyperactivity in the open field which was maximal at 20–25 days of age and declined towards normal values at 40 days of age. At maturity, between 60 and 80 days of age, these rats showed poor performance in a complex 6-choice discrimination learning but not in simple differential conditioning. Possible correlations between the behavioural and biochemical findings are discussed.

Fasting reduces cardiac beta-adrenergic receptor concentration and ornithine decarboxylase activity in rats

Fasting reduces cardiac beta-adrenergic receptor concentration and ornithine decarboxylase activity in rats

The increase in hormone-stimulated adenylate cyclase activity following Rous sarcoma virus transformation.

Rous sarcoma virus (RSV)-infected chicken embryo cells were used to study the effect of viral transformation on the hormone-stimulated synthesis of cyclic AMP. Transformation by RSV greatly increased the cells' ability to synthesize and accumulate cyclic AMP in response to the beta-adrenergic agonist isoproterenol as compared to untransformed cells. This enhancement was observed in both intact cells and in membranes prepared from these cells. The inclusion of guanosine 5'-0-(3-thiotriphosphate), a nonhydrolyzable analogue of GTP, in assays of adenylate cyclase activity did not abolish the quantitative differences between the transformed and normal cell membranes. Infection of cells by Rous-associated virus, which lacks the oncogene src, did not induce this hyperresponsiveness thus indicating the probable involvement of the src gene product in this phenomenon. The duration of the isoproterenol-induced cyclic AMP elevation was longer in the transformed than in the untransformed cells; transformed cells, unlike untransformed cells, required at least 120 min before full desensitization became established. Membranes prepared from transformed cells specifically bound more than 5 times the quantity of the beta-adrenergic radiolabeled antagonist (-)3H-dihydroalprenolol and 125I-iodocyanopindolol compared to the untransformed cell membranes. Thus, it appears that major differences between the transformed and normal phenotypes reside in the concentration of membrane beta-adrenergic receptors and the inability of RSV-transformed cells to self-limit their response to specific external stimuli.

Receptor-associated changes of the catecholamine-sensitive adenylate cyclase in glioma cells doubly transformed with Moloney sarcoma virus.

A doubly transformed rat glioma cell line, designated C6V-1, was obtained from rat glioma C6 cells by infection with a rat-adapted variant of Moloney sarcoma virus (MSV-M-os). The C6V-1 cells show karyotypic changes in chromosome number (43) and structure, while C6 cells possess a normal male karyotype. C6V-1 and C6 cells were employed for characterization of a receptor-adenylate cyclase system of the surface membrane. C6V-1 cells showed lower adenylate cyclase activity than that of C6 cells, though the apparent Km for ATP in both types of cells was the same. The maximal stimulation of adenylate cyclase by isoproterenol was significantly reduced, and Kact for isoproterenol was approximately 18-fold lower in C6V-1 cells. When the concentration of beta-adrenergic receptors was measured by various concentrations of [3H] dihydroalprenolol (DHA), the maximal binding sites of C6 and C6V-1 cells were 760 and 230 fmol/mg protein, respectively, without any changes in the association constant for DHA. The concentration of isoproterenol required for 50% displacement of the [3H] DHA binding (Kd) was the same (around 1.5 X 10(-6)M) in both cells, measured in the presence of GTP. Thus the 19-fold drop in the Kd/Kact ratio in C6V-1 cells suggests an incomplete coupling of beta-receptors to adenylate cyclase. Cyclic AMP phosphodiesterase activity and cAMP content in C6V-1 were lower than in C6 cells. Mitochondrial monoamine oxidase and cytosomal enolase activities, however, were somewhat higher in C6V-1 cells. The results indicate that a set of changes in the receptors and in the cyclic AMP system of C6V-1 is one of the specific alterations by transformation, even though those may not be the cause of cell transformation.

Possible involvement of parotid beta-adrenergic receptors in the etiology of sialadenosis.

The concentration of beta-adrenergic receptors was determined in rat and human parotid glands, in normal tissue as well as after sympathetic denervation of the rat, and in human sialadenosis. Receptor levels were clearly elevated after denervation of the rat and in sialadenosis. The possible implications of these findings for the etiology of human sialadenosis are discussed.

The reversal of the Gpp(NH)p-activated state of adenylate cyclase by GTP and hormone is by the “collision coupling” mechanism.

The guanylyl imidodiphosphate-activated state of turkey erythrocyte cyclase can be reversed to the basal state by the simultaneous action of beta-agonists and GTP. The rate of reversal diminishes progressively with decreasing concentration of beta-adrenergic receptors on the membrane surface, whereas the extent of reversal is always maximal. The rate of reversal is found to be linearly dependent on the concentration of beta-adrenergic receptors within the membranes. This result supports the notion that the interaction of the enzyme unit and the beta-adrenergic receptor is catalytic and therefore of the "collision coupling" type. The dependence of the rate of reversal reaction on epinephrine concentration is noncooperative with an apparent dissociation constant of KD = 3.0 X 10(-6) M. The fraction of guanylyl imidodiphosphate-activated cyclase system which can be reversed by GTP and a beta-agonist strongly depends on temperature and reveals a sharp transition at 24 degrees C which is the freezing temperature of the inner monolayer. It is suggested that the GTP regulatory unit is inactive when the inner monolayer is frozen.

Glucocorticoids increase pulmonary beta-adrenergic receptors in fetal rabbit.

beta-Adrenergic agonists stimulate surfactant release and decrease fluid in lung alveoli of fetuses. Both effects are most evident toward the end of gestation. We used [3H] dihydroalprenolol (DHA) to investigate the development of pulmonary beta-adrenergic receptors in rabbit fetuses and to study the effect of glucocorticoid treatment on the beta-receptor number. In the lung particulate preparation, DHA binding was rapid, reversible, stereoselective, and of high affinity. The order of potency for adrenergic agonists in competing for DHA binding was isoproterenol > epinephrine = norepinephrine, which is typical of interactions at a beta 1-adrenergic receptor. Using DHA, we demonstrated that the concentration of pulmonary beta-receptors increased significantly between 28 and 31 days of gestation; however, there was no change in the dissociation constant during gestation. After injecting betamethasone (0.17 mg/kg, 24 hours) into rabbits at 25 days of pregnancy, we found that the concentration of pulmonary beta-receptors increased from 44.2 +/- 6.6 fmol/mg protein in untreated fetuses to 77.9 +/- 5.6 fmol/mg protein in treated fetuses. However, this treatment did not affect the DHA binding sites in the fetal rabbit heart. Maternal treatment with the T3 analogue 3,5-dimethyl-3'-isopropyl-L-thyronine (0.5-1 mg/kg) at a dosage which increased both surfactant synthesis and release did not alter pulmonary receptor concentration. Our results indicate that the concentration of pulmonary beta-adrenergic receptors increases in the fetus at term and suggest that this increase is stimulated by endogenous glucocorticoid in fetal circulation.

The human placenta--a rich source of beta-adrenergic receptors: characterization of the receptors in particulate and solubilized preparations.

A crude particulate fraction of human placenta possesses a high concentration of beta-adrenergic receptors, as determined by (-)-[3H]dihydroalprenolol binding (approximately 240 fmol/mg protein; Kd approximately 2 nM). The sites display all the typical characteristics of beta-adrenergic receptors, including rapid and reversible kinetics, saturability, and appropriate specificity and stereospecificity. Computer modelling of ligand binding data indicate that the binding of (-)-[3H]dihydroalprenolol to these sites conforms closely to the pattern anticipated for interactions of the ligand with a homogeneous class of receptors according to the law of mass action. The rejeptors are readily solubilized with digitonin, retaining their typical beta-adrenergic characteristics. The human placenta is likely to be a particularly useful source of beta-adrenergic receptors for purification because of its high receptor content and its ready availability in substantial quantities.

Reduced vascular beta-adrenergic receptors in deoxycorticosterone-salt hypertensive rats

A method has been established for measuring beta-adrenergic receptors in membranes prepared from rat mesenteric arteries. The beta-adrenergic antagonist 125 I-iodohydroxybenzylpindolol was used as ligand in direct binding experiments. In the presence of 10 −4M phentolamine, 125I-iodohydroxybenzylpindolol binding was saturable, stereospecific, rapid and reversible. The relative potencies of beta-adrenergic agonists in displacing 125I-iodohydroxybenzylpindolol correlated with their relative potencies in stimulating adenylate cyclase activity in artery homogenates. This method was used to measure beta-receptors in mesenteric arteries of rats made hypertensive by deoxycorticosterone-salt treatment. Arteries from rats with established hypertension contained a reduced concentration of betaadrenergic receptors. This decrease was from 10.1 ± 1.5 (S.E.M.) to 7.0 ± 1.2 fmoles/mg protein. There was no change in affinity for 125I-iodohydroxybenzylpindolol (0.10 ± 0.02n M).

Beta adrenergic receptors in lymphocyte subpopulations

To further evaluate the potential utility of lymphocyte beta adrenergic receptor assays in the study of receptor alterations in human disease, we studied highly purified populations of B and T lymphocytes in peripheral blood to see if differences existed in the concentration or affinity of beta adrenergic receptors and catecholamine-responsive cAMP levels. The mean number of receptors present in particulate fractions of B cells did not differ significantly from the number found in T cells. Similarly, no significant difference in the dissociation constant for (−)[ 3H]dihydroalprenolol was found. Cyclic adenosine monophosphate (cAMP) accumulation in whole lymphocytes as measured by radioimmunoassay was comparable, although a tendency toward lower basal and stimulated levels in the T cells was evident. The data suggest that differences observed in concentrations of beta adrenergic receptors or catecholamine-responsive cAMP accumulation in lymphocytes from patients with varying illnesses are not likely to be due to differences in the proportions of circulating B and T lymphocytes.