Adrenergic Receptor Binding Research Articles

Changes in Platelet Alpha-2-Adrenoceptors in Fibromyalgia: Effects of Treatment with Antidepressants

The aim of this study was to determine platelet α₂-adrenergic receptor (α₂-AR) binding sites in fibromyalgia both before and after treatment with sertraline or placebo. The maximum number of binding sites (B_max) and their affinity (K_d) for [³H]rauwolscine, a selective α₂-AR antagonist, were measured in 13 normal volunteers and 22 fibromyalgia patients. Severity of illness was evaluated by means of the Hamilton Depression Rating Scale (HDRS) and dolorimetric assessments of tenderness at tender points. Fibromyalgia patients had repeated measurements of [³H]rauwolscine binding characteristics both before and after subchronic treatment with sertraline or placebo for 12 weeks. [³H]rauwolscine binding K_d values were significantly higher in fibromyalgia patients than in normal volunteers. There were significant inverse correlations between [³H]rauwolscine binding K_d values and duration of illness, age and lower energy. Significantly higher [³H]rauwolscine binding K_d values were found in fibromyalgia patients in an early phase of illness (<3 years) than in fibromyalgia patients with a protracted illness (>3 years). Repeated administration of sertraline had no significant effects on [³H]rauwolscine binding B_max or K_d values. The results suggest that fibromyalgia and, in particular, fibromyalgia in an early phase of illness, is accompanied by lowered affinity of platelet α₂-ARs.

Subtype-selective N-methyl-D-aspartate receptor antagonists: synthesis and biological evaluation of 1-(arylalkynyl)-4-benzylpiperidines.

A search of our compound library for compounds with structural similarity to ifenprodil (5) and haloperidol (7) followed by in vitro screening revealed that 4-benzyl-1-(4-phenyl-3-butynyl)piperidine (8) was a moderately potent and selective antagonist of the NR1A/2B subtype of NMDA receptors. Substitution on the benzyl group of 8 did not significantly affect NR1A/2B potency, while addition of hydrogen bond donors in the para position of the phenyl group enhanced NR1A/2B potency. Addition of a hydroxyl moiety to the 4-position of the piperidine group slightly reduced NR1A/2B potency while reducing alpha-1 adrenergic and dopamine D2 receptor binding affinities substantially, resulting in improved overall selectivity for NR1A/2B receptors. Finally, the butynyl linker was replaced with propynyl or pentynyl. When the phenyl was para substituted with amine or acetamide groups, the NR1A/2B potency order was butynyl > pentynyl >> propynyl. For the para methanesulfonamide or hydroxyl groups, the order was butynyl approximately propynyl > pentynyl. The hydroxyl propyne (48) and butyne (23) were among the most potent NR1A/2B antagonists from this study. They both potentiated the effects of L-DOPA in the 6-hydroxydopamine-lesioned rat, a model of Parkinson's disease, dosed at 10 mg/kg ip, but 48 was not active at 30 mg/kg po.

Kainic acid (KA)-induced seizures in Sprague-Dawley rats and the effect of dietary taurine (TAU) supplementation or deficiency.

Male Sprague-Dawley rats received TAU supplementation (1.5% in drinking water) or TAU deficient diets for 4 weeks to test for a possible neuroprotective role of TAU in KA-induced (10 mg/kg s.c.) seizures. TAU supplementation significantly increased serum and hippocampal TAU levels, but not TAU content in temporal cortex or striatum. TAU deficient diets did not attenuate serum or tissue TAU levels. Dietary TAU supplementation failed to decrease the number or latency of partial or clonic-tonic seizures or wet dog shakes, whereas a TAU deficient diet decreased the number of clonictonic and partial seizures. This study does not support previous observations of an anticonvulsant effect of TAU against KA-induced seizures. KA-treatment decreased alpha 2-adrenergic receptor binding sites and TAU content in the temporal cortex across all dietary treatment groups, supporting previous evidence of severe KA-induced damage and neuronal loss in this brain region.

Dihydropyridine and beta adrenergic receptor binding in dogs with tachycardia-induced atrial fibrillation.

We have shown that rapid atrial activation, as occurs during atrial fibrillation (AF), reduces L-type Ca2+ current (ICa) and that this is the principal mechanism of the action potential duration and refractoriness changes that characterize tachycardia-induced atrial remodeling. The present study was designed to determine whether atrial tachycardia alters biochemical indices of the number of L-type Ca2+ channels and/or of the number and binding affinity of beta-adrenergic receptors. In canine atrial sarcolemmal preparations, the number and binding affinity of dihydropyridine receptors were determined with the use of 3H-nitrendipine and that of beta-adrenergic receptors with 125I-iodocyanopindolol. Results were obtained with preparations from dogs paced at 400/min for 1 (P1, n = 20), 7 (P7, n = 9), and 42 (P42, n = 9) days, and compared with observations in sham-operated controls (P0, n = 14). Pacing reduced the Bmax of dihydropyridine receptors, from 157 +/- 18 fmol/mg (P0) to 116 +/- 9 fmol/mg (P1, P < 0.05), 100 +/- 14 fmol/mg (P7, P < 0.05) and 94 +/- 9 fmol/mg (P42, P < 0.01). The affinity of dihydropyridine receptors was unchanged, with the Kd averaging 711 +/- 102 pM. 656 +/- 74 pM, 633 +/- 155 pM and 585 +/- 92 pM in P0, P1, P7 and P42 dogs. Neither Bmax nor Kd of beta-adrenergic receptors was altered by rapid pacing. Values of Bmax of dihydropyridine receptors correlated with atrial ICa current density (r2 = 0.95) and ERP (r2 = 0.99). Rapid atrial activation results in downregulation in the number of dihydropyridine receptors without altering the number or affinity of beta-adrenergic receptors. The reductions in ICa that play an important role in the atrial electrical remodeling by which 'AF begets AF' appear to be due at least in part to a decrease in the number of L-type Ca2+ channels in cardiac cell membranes.

Adrenergic receptors in premenstrual dysphoric disorder. II. Neutrophil β 2-adrenergic receptors: G s protein coupling, phase of menstrual cycle and prediction of luteal phase symptom severity

Abnormal β 2-adrenergic receptor coupling to G s protein is implicated in depressive disorders. Steroid hormones and antidepressants modulate β-adrenergic receptor coupling, which may relate to the therapeutic efficacy of antidepressants. We examined β 2-adrenergic receptors in 18 patients with premenstrual dysphoric disorder (PMDD), in 15 control subjects during the follicular phase and in 12 patients during late luteal phase. Antagonist-measured receptor density, agonist-measured receptor density in the high- and low-conformational states and agonist affinity to both states were measured. Coupling indices to G s protein were determined from agonist-displacement experiments. Follicular β 2-adrenergic receptor density was higher in patients than in control subjects, with a trend for higher receptor density in the high-conformational state. The phase of menstrual cycle had no effect on β 2-adrenergic receptor regulation in PMDD. Exploratory correlations showed that the K L/ K H ratio was related to anxiety ratings in control subjects and % R H was correlated with symptom severity in patients. In patients, follicular β 2-adrenergic receptor binding measures were correlated with luteal symptom severity. These findings suggest abnormal β 2-adrenergic receptor regulation in PMDD. Further exploration of the role of β-adrenergic receptor kinase, sex steroid hormones and antidepressants on β-adrenergic receptor regulation in PMDD is warranted.

Hypothyroidism-evoked shifts in hippocampal adrenergic receptors: implications to ischemia-induced hippocampal damage.

Hypothyroidism was induced in a group of male Fischer 344 rats by administration of 0.05% propylthiouracil (PTU) in the drinking water for 12 weeks. Control rats were not treated. Plasma levels of thyroid hormones indicated that PTU treatment had produced severe thyroid hormone deficiency. In PTU-treated rats compared to control rats, levels of total T3 and total T4 were reduced 54.5% and 53.7%; while levels of free T3 and free T4 were reduced 87.1% and 96.5%. Functional hypothyroidism was demonstrated by: (i) a 49.1% decrease in hepatic plasma membrane alpha1-adrenergic receptor binding, and (ii) a 11.2-fold increase in hepatic gamma-glutamyltranspeptidase activity; relative to the expression of these parameters in control rats. Membranes were isolated from hippocampi of control, PTU-induced hypothyroid and thyroxine-replaced rats and specific adrenergic receptor binding determined by radioligand binding techniques. Hypothyroidism resulted in a shift in the balance of alpha1 and beta2 adrenergic receptor binding by evoking: an increase in alpha1-adrenergic receptor binding to 1.57-fold of control levels; and, a decrease in beta2-adrenergic receptor binding to 64% of control levels. Thyroid hormone replacement carried out in PTU-treated hypothyroid rats at 30 microg/kg s.c. per day for the last 3 days of the 12 week PTU-treatment protocol, which reversed physical and functional hypothyroidism, reversed the observed changes in hippocampal adrenergic receptor binding, indicating them to be thyroid hormone, and not PTU, -dependent. This receptor shift evoked by hypothyroidism may, in part, explain the protective effect of hypothyroidism on ischemia-induced hippocampal damage by favoring inhibitory input and limiting excitotoxic input by catecholamines.

Effects of monocular enucleation on receptor binding and innervation pattern of the noradrenergic system in the superior colliculus of the pigmented rat

To examine visual activity-dependent regulation of the central noradrenergic (NAergic) system, we carried out in vitro autoradiography for α 2-, β-adrenergic and NMDA, non-NMDA glutamatergic receptors together with immunohistochemical analysis for dopamine β-hydroxylase in the major central visual structures, including the lateral geniculate nucleus, superior colliculus (SC), and visual cortex of pigmented rats, that had received monocular enucleation. β-Adrenergic receptor ( β-AR) binding in the SC contralateral to the enucleated eye was significantly decreased (82%, P<0.01) following monocular deprivation started at P12 and continued for 2 or 11 weeks. No significant change in β-AR binding was found in other structures examined. The number of varicosities in NAergic fibers was significantly increased following longer enucleation, i.e. for 11 weeks, in the contralateral SC (197%, P<0.001), whereas that in the ipsilateral SC was reduced (75%, P<0.001). Changes in α 2-adrenergic, NMDA, and non-NMDA glutamatergic receptor binding were small in these animals. Changes in neither β-AR binding nor innervation pattern of NAergic fibers were found in one-year-old rats that had received a comparable period of monocular enucleation. Furthermore, neither unilateral ablation of the visual cortex to reduce a different set of major afferents nor neonatal enucleation, which induced anatomical reorganization of the afferents, was found to be effective. These findings suggest that β-AR binding and innervation pattern of NAergic fibers in the SC are modified only when massive imbalance of retinal afferent activity is imposed during a limited period in early postnatal life (i.e. the sensitive period).

Carboxyl-promoted enhancement of selectivity for the β 3 adrenergic receptor. Negative charge of the sulfonic acid BMS-187413 introduces-β 3 binding selectivity

Carboxyl and other negatively charged groups were found to be most effective at producing human β 3 adrenergic receptor binding selectivity in 1 (BRL 37344) and related compounds. The sulfonic acid analog 7 (BMS-187413) is a novel and potent β 3 adrenergic agonist that binds selectively, and thus has an in vitro profile that compares favorably with that of BRL 37344.

Down-regulation of β-adrenergic receptor following long-term monocular deprivation in cat visual cortex

To examine how adrenergic receptor binding is modified by experimental manipulation of sensory afferent, we carried out binding experiments (membrane fraction and in vitro autoradiography) for both α 2- and β-adrenergic receptors in the brain of cats which had been deprived of vision in one eye. In the cerebral cortex of control animals, β-adrenergic receptor (β-AR) binding was found to be higher in the occipital regions than in other regions, while α 2-AR binding was relatively uniform. Monocular deprivation throughout the postnatal sensitive period (1–7 month of age) significantly decreased β-AR binding in the visual cortex and lateral geniculate nucleus. Scatchard plot analysis in the visual cortex showed ca. 50% reduction in B max and little change in K d No significant difference was found in α 2-AR binding following monocular deprivation. Similar extent of down-regulation in β-AR binding was confirmed in all layers of visual cortex using autoradiography.

Chiral recognition in adrenergic receptor binding mimics prepared by molecular imprinting.

Molecularly imprinted polymers were prepared against the adrenomimetic agents ephedrine and pseudoephedrine. These compounds each incorporate two chiral centres. The polymers were evaluated with respect to enantiodiscrimination of various adrenergic ligands. The selectivity of the polymeric binding sites for the imprinted molecules was very high, and it was found that binding of both the enantiomeric and diastereomeric isomers of the imprint species were effectively obstructed, it was found that these polymers could selectively recognize the enantiomers of the endogenous adrenergic ligand epinephrine as well as several beta-adrenergic blockers. These observations suggest that these polymers effectively mimic the recognition patterns exhibited by natural adrenergic receptors.

Noradrenergic mechanisms in stress and anxiety: II. Clinical studies

Studies in animals have shown a relationship between alterations in noradrenergic brain system function and behaviors of anxiety and fear. These findings have generated the hypothesis that the symptoms seen in patients with anxiety disorders may be related to alterations in noradrenergic function. A number of clinical studies have tested this hypothesis, utilizing measures of catecholaminergic function such as heart rate and blood pressure, measurement of norepinephrine and its metabolites in urine and plasma and adrenergic receptor binding in platelets, as well as pharmacological challenge to the noradrenergic system. Acute stressors, such as public speaking, have been associated with an increase in heart rate, blood pressure, and norepinephrine and its metabolites in urine and plasma. Findings in patients with panic disorder at baseline related to heart rate, blood pressure, baseline norepinephrine and its metabolites, and platelet adrenergic receptors have been mixed, while the most consistent findings have been blunted growth hormone response to clonidine and increased 3-methoxy-4-hydroxy-phenylethylene-glucol (MHPG) and anxiety following stimulation of the noradrenergic system with yohimbine. Baseline measures of noradrenergic function in patients with posttraumatic stress disorder (PTSD) have also been mixed, while an increased heart, blood pressure and norepinephrine response to traumatic reminders, as well as increased behavioral (as well as different brain metabolic) response to yohimbine, have been found in PTSD. There are fewer studies of noradrenergic function in the other anxiety disorders, and the findings there have not been consistent. These studies provide evidence for increased noradrenergic responsiveness in panic disorder and PTSD, although there does not appear to be an alteration in baseline noradrenergic function in these patients. © 1996 Wiley-Liss, Inc.

Noradrenergic mechanisms in stress and anxiety: II. Clinical studies

Studies in animals have shown a relationship between alterations in noradrenergic brain system function and behaviors of anxiety and fear. These findings have generated the hypothesis that the symptoms seen in patients with anxiety disorders may be related to alterations in noradrenergic function. A number of clinical studies have tested this hypothesis, utilizing measures of catecholaminergic function such as heart rate and blood pressure, measurement of norepinephrine and its metabolites in urine and plasma and adrenergic receptor binding in platelets, as well as pharmacological challenge to the noradrenergic system. Acute stressors, such as public speaking, have been associated with an increase in heart rate, blood pressure, and norepinephrine and its metabolites in urine and plasma. Findings in patients with panic disorder at baseline related to heart rate, blood pressure, baseline norepinephrine and its metabolites, and platelet adrenergic receptors have been mixed, while the most consistent findings have been blunted growth hormone response to clonidine and increased 3-methoxy-4-hydroxy-phenylethylene-glucol (MHPG) and anxiety following stimulation of the noradrenergic system with yohimbine. Baseline measures of noradrenergic function in patients with posttraumatic stress disorder (PTSD) have also been mixed, while an increased heart, blood pressure and norepinephrine response to traumatic reminders, as well as increased behavioral (as well as different brain metabolic) response to yohimbine, have been found in PTSD. There are fewer studies of noradrenergic function in the other anxiety disorders, and the findings there have not been consistent. These studies provide evidence for increased noradrenergic responsiveness in panic disorder and PTSD, although there does not appear to be an alteration in baseline noradrenergic function in these patients.

α2-Adrenergic receptor binding in canine Purkinje fibers

Autoradiograms were developed from free running strands of Purkinje fibers and from left ventricular myocardium of dog hearts exposed to [7- methoxy- 3H]prazosin or [ O- methyl- 3H]yohimbine in the presence or absence of excessive non-radioactive ligands. Both Purkinje fiber and ventricular myocardium showed high density, tissue-specific binding to [ 3H]prazosin. In contrast, high density, tissue-specific binding to [ 3H]yohimbine was present in Purkinje fibers but not in ventricular myocardium. Membrane fractions showed high affinity, saturable, and displaceable binding with [ 3H]yohimbine in preparations from canine cardiac Purkinje fibers but not those from canine cardiac ventricular myocardium. Scatchard analysis of the canine Purkinje membrane α 2-adrenergic receptor binding showed a B max of 54.9 fmol/mg protein with a K d of 6.25 nM. These results confirm the electrophysiological findings that post-junctional α 2-adrenergic receptors are present in Purkinje fibers.

Effect of beef tallow diet on alpha 2-adrenergic receptor binding in rat brain regions.

We examined the effects of dietary fats consisting of different fatty acids on alpha 2-adrenergic receptor binding in the hypothalamus and cerebral cortex. Sprague-Dawley male rats were meal-fed isoenergetic diets based on safflower oil or beef tallow for 8 weeks. alpha 2-Adrenergic receptor bindings were determined with [3H]para-aminoclonidine. Binding affinities of alpha 2-adrenergic receptor in the hypothalamus and cortex were significantly higher in the beef tallow diet group. The values of the polyunsaturated/saturated fatty acid ratio and the fluidity of the plasma membrane in the hypothalamus and cortex were lower in the beef tallow diet group than in the safflower oil diet group. These results suggest that the beef tallow diet decreases membrane fluidity by altering the fatty acid composition in rat brain regions. Consequently, alpha 2-adrenergic receptor binding affinities in the brain regions are higher in rats fed the beef tallow diet than in rats fed the safflower oil diet.

Benzodiazepine agonists reverse the effects of noise exposure on central benzodiazepine receptors and cardiac responsiveness

Rats were submitted to 110 dB white noise exposure for 1, and 6 hours and brain α 1, β 1 and benzodiazepine receptor binding was evaluated with selective ligands. An increase in cerebral benzodiazepine receptor (CBR) concentration, without any significant change in affinity constant, occurred after the 6 h treatment; no change was observed in adrenergic receptor binding at any period of exposure. Both diazepam and clonazepara pre-treatment reversed the effects of noise on CBR binding, confirming a role of these receptors in the response to noise stress. Furthermore, these benzodiazepine agonists influenced the response of cardiac and aortic tissues, which are known to be changed by stress exposure. Diazepam and clonazepam pre-treatment protected cardiac tissue from the effects of 6h noise stress, and a potentiation of aortic responses was detected, although at different tunes of exposure. The differences between the responses of these peripheral tissues to benzodiazepine treatment suggest that the expression depends on the tissue examined and the period of exposure.

Tannin inhibits adenylate cyclase in airway epithelial cells

Tannin, isolated from cotton bracts extract and implicated in the pathogenesis of byssinosis, inhibits adenosine 3',5'-cyclic monophosphate (cAMP) production and Cl- secretion in bovine airway epithelial cells in part by inhibiting adrenergic receptor binding. The purpose of this study was to determine whether tannin affected other parts of the adrenergic-cAMP signal transduction pathway by examining the effect of tannin on guanosine 5'-triphosphate (GTP)-regulatory pathways (G proteins) and on adenylate cyclase activity. cAMP production in confluent airway epithelial cells was measured in the presence of cholera toxin (100 micrograms/ml), an activator of GS proteins, and forskolin (0.1-1,000 microM), a direct activator of adenylate cyclase. Cholera toxin stimulated cAMP production; this response, however, was inhibited in cells pretreated with 50 micrograms/ml tannin. Forskolin (100 microM) stimulated cAMP production 13-fold above baseline values. Tannin pretreatment inhibited the stimulatory effect of forskolin on cAMP release in a dose-dependent manner with a tannin concentration causing 50% inhibition of 7.5 micrograms/ml. The stimulatory effect of forskolin on cAMP release was completely inhibited in cells pretreated with 50 micrograms/ml tannin. The inhibition was reversible 3 h after removal of tannin from the solution. Tannin also inhibited forskolin-stimulated adenylate cyclase activity in a dose-dependent, noncompetitive manner. We conclude that forskolin and cholera toxin stimulate cAMP production in airway epithelial cells and that tannin inhibits the production of cAMP in airway epithelial cells by a direct effect on adenylate cyclase activity.

Palmitoylation of the alpha 2A-adrenergic receptor. Analysis of the sequence requirements for and the dynamic properties of alpha 2A-adrenergic receptor palmitoylation.

Previous studies have demonstrated that the alpha 2A-adrenergic receptor (alpha 2A AR) incorporates [3H]palmitate and that replacement of Cys442 by Ala or Ser eliminates detectable acylation without perturbing coupling to pertussis toxin-sensitive GTP-binding proteins (Kennedy, M. E., and Limbird, L. E. (1993) J. Biol. Chem. 268, 8003-8011) or, as shown here, without perturbing agonist-dependent receptor phosphorylation, in contrast to the consequences of eliminating beta 2-adrenergic receptor acylation. As a first step in revealing the functional role for this post-translational modification at the alpha 2A AR, we explored sequences in the alpha 2AAR which confer alpha 2AAR acylation and whether or not [3H]palmitoylation of the alpha 2AAR is dynamic. Deletion of the 7 terminal amino acids distal to Cys442 of the alpha 2AAR did not eliminate detectable [3H]palmitoylation of the alpha 2AAR, whereas truncation to Leu441 did, indicating both that Cys442 is the likely site for acylation and that sequences distal to Cys442 are not required for acylation at Cys442. Since mutation of sequences proximal to Cys442 altered overall receptor structure, based on markedly reduced detectable adrenergic receptor binding, proximal motifs required for palmitoylation of the alpha 2AAR could not be explored further. When the turnover of [35S]Met/Cys-labeled alpha 2AAR was compared with the turnover of the [3H]palmitate-labeled alpha 2AAR, it was of interest that agonist treatment accelerated the half-life of decay of the [3H]palmitate-labeled alpha 2AAR without detectable receptor down-regulation, providing evidence that the acylation of the alpha 2AAR may be a dynamic process.

Liver beta-adrenergic receptors, G proteins, and adenylyl cyclase activity in obesity-diabetes syndromes

The ob and db genes produce similar hormonal anomalies in mice. Although the expression of the syndromes diverges with age, at 8-12 wk both ob/ob and db/db mice are hyperglycemic and hyperinsulinemic and show evidence of hypercorticoidism. Nevertheless, membranes isolated from livers of ob/ob and db/db mice behave differently in terms of adenylyl cyclase activity and beta-adrenergic receptor function. There are three times as many beta 2-adrenergic receptor binding sites and a threefold increase in the response to catecholamines in ob/ob mouse liver membranes than in comparable preparations from normal controls or db/db mice. By contrast, the two main G proteins of liver membranes (Gs alpha and Gi alpha 2) are less abundant in the mutants, ob/ob and db/db, than in their respective lean controls. Adrenalectomy normalizes the exaggerated response to beta-adrenergic agonists and the number of beta-adrenergic binding sites in the ob/ob mouse. This shows that the enhanced beta-adrenergic receptor response is linked to hypercorticoidism. Cellular maturation and differentiation (D. C. Watkins, J. K. Northrup, and C. C. Malbon, J. Biol. Chem. 262: 10651-10657, 1987) and diseases such as obesity and diabetes (cf. N. McFarlane-Anderson, J. Bailly, and N. Bégin-Heick, Biochem. J. 282: 15-23, 1992) have been associated with modifications in the complement of G proteins detected in cells. However, the relationship among levels, types, and intracellular localization of G proteins in tissues and their influence on the transduction of the message to an effector system, such as adenylyl cyclase, are not yet well understood.

Dose-effect and pharmacokinetic-pharmacodynamic relationships of the β1adrenergic receptor blocking properties of various doses of carvedilol in healthy humans

To evaluate the pharmacodynamic properties of carvedilol across a broad range of doses in relation to its enantiospecific kinetics and adrenergic receptor occupancies, relative to placebo and propranolol. Twelve healthy male subjects were investigated on six separate occasions at least 1 week apart when they received either a single peroral dose of 40 mg propranolol, 12.5, 25, 50, or 100 mg carvedilol, or placebo. The subjects were extensively profiled at supine rest, and they underwent supine bicycle ergometry before and at 2, 4, 6, 9, 12, and 22 hours after dosing. At these time points blood was drawn for the high performance liquid chromatographic determination of the enantiomers of carvedilol and for the radioreceptor assay determination of alpha 1- and beta 1-adrenergic receptor binding and related concentrations. Carvedilol was confirmed to bind to beta 1-adrenergic receptors and (albeit to a lesser extent) to alpha 1-adrenergic receptors. Carvedilol furthermore attenuated the ergometric increase in heart rate in a closely dose-related fashion, which exemplified its beta 1-adrenergic receptor blocking effects. However, the basal efferent adrenergic drive might have been too low to show consistent alpha 1-blocking properties. The radioreceptor and enantiomer kinetics were proportional with dose. There was no indication that the overall kinetic behavior of contributing active metabolites would differ from that of the S(-)-enantiomer. On average, there was a smooth linear relationship between the ergometric treatment responses and log-transformed dose, log-transformed concentrations of the S(-)-enantiomer, and the radioreceptor assay derived beta 1-adrenergic receptor occupancies. The relative complexity of the kinetics of carvedilol (enantiospecific kinetics and dynamics, protein binding, and involvement of active metabolites) does not preclude relatively simple and straight-forward dose-effect and kinetic-dynamic relationships.

Distribution and density of α– and β–adrenergic receptor binding sites in the bovine mammary gland

Radioreceptor binding studies were designed to localize and determine the number of alpha- and beta-adrenergic receptors in the mammary gland of lactating cows. 3H-prazosin, 3H-rauwolscine and 3H-dihydroalprenolol were used for the regional characterization of alpha 1-, alpha 2- and beta-adrenergic receptors by competitive inhibition of binding of 3H-ligands with unlabelled adrenergic agonists and antagonists. The alpha 1-, alpha 2- and beta 2-adrenergic receptor subtypes could thus be demonstrated in the regions of the teats, large mammary ducts and parenchyma. Tissues of the teat wall, of the large mammary ducts above the gland cistern and of the mammary parenchyma were prepared to determine the density of alpha 1-, alpha 2- and beta-receptors by saturation binding assays using 3H-prazosin, 3H-rauwolscine and 3H-dihydroalprenolol respectively. Binding to high affinity sites was reversible within minutes and saturable. Equilibrium was reached within minutes. The number of alpha 1- and alpha 2-adrenergic receptors decreased from the teat to the mammary ducts to the parenchyma. Most of the alpha 1- and alpha 2-adrenergic receptors were found in the teat wall, whereas in the parenchyma alpha-adrenergic receptors were absent or barely detectable. The density of beta-adrenergic receptors was similar in the teat wall and the large mammary ducts, but much lower in the parenchyma. Thus, alpha 1-, alpha 2- and beta-adrenergic receptors were found mainly in the milk purging system and hardly at all in mammary parenchyma. Inhibition of milk removal by alpha-adrenergic stimulation is possibly due to constriction of teat wall and to constriction of the mammary ducts, whereas enhanced milk flow after beta-adrenergic stimulation is possibly due to relaxation not only of the teat sphincter and teat wall, but probably also of the large mammary ducts.