Inversal relationship between basal adenylcyclase activity and maximal degree of stimulation in membranes of red blood cells from rats

Summary The degree of stimulation of adenylcyclase activity, in membranes from immature red blood cells from rats, brought about by isoprenaline, guanylyl-imidodiphosphate and sodium fluoride is strongly dependent on the basal activity of the enzyme. The inversal relationship between basal activity and the maximal degree of stimulation by (–) isoprenaline, shows an apparent seasonal dependence.This work was supported by a grant from the Deutsche Forschungsgemeinschaft.     

Opposite effects of beta-adrenoceptor stimulation and 8-bromo-cyclic AMP on potassium efflux in mammalian heart muscle

beta-adrenoceptor stimulation by isoprenaline increases the potassium efflux in beating guinea-pig atria. This effect is not mimicked by 8-bromo-cyclic AMP, a cyclic AMP analogue which exerts a positive inotropic effect in this preparation.     

Opposite effects ofβ-adrenoceptor stimulation and 8-bromo-cyclic AMP on potassium efflux in mammalian heart muscle

Summary -adrenoceptor stimulation by isoprenaline increases the potassium efflux in beating guinea-pig atria. This effect is not mimicked by 8-bromo-cyclic AMP, a cyclic AMP analogue which exerts a positive inotropic effect in this preparation.     

An analysis of the effects of urethane on cardiovascular responsiveness to catecholamines in terms of its interference with Ca++ mobilization from both intra and extracellular pools

Urethane (1 X 10(-2) - 1 X 10(-1) M) reduced, in a concentration-dependent manner, both intra and extracellular Ca++ dependent noradrenaline-induced contractions of perfused rabbit ear artery as well as the tonic contractions produced by perfusion with high K+ solution. However, a quantitative analysis of the data indicated that for urethane concentrations similar to those found in plasma during anesthesia urethane antagonism is confined to noradrenaline-induced contractions which depend upon the mobilization of Ca++ from intracellular storage sites. In KCl-contracted arteries, urethane enhanced the relaxant effects of isoprenaline. - Urethane reduced the amplitude of contractions of spontaneously beating guinea-pig right atrium at concentrations which have only a limited effect on frequency. In addition, it decreased in a concentration-dependent manner the amplitude of isoprenaline-activated electrically driven, and K+ depolarized guinea-pig right ventricular strips. Urethane had no effect on the chrono and inotropic actions of isoprenaline on cardiac preparations. In in vivo experiments the chronotropic response to low doses of isoprenaline was significantly higher in urethane-treated as compared to unanesthetized rats. The higher dose of isoprenaline tested produced a significant fall in systolic blood pressure in urethane-anesthetized rats. A significant correlation exists between the chronotropic response to isoprenaline and resting heart rate values in urethane-anesthetized rats. These results indicate that urethane, at concentrations similar to those found in plasma during anesthesia selectively interferes with mobilization of Ca++ from intracellular storage sites. In addition, the interference of urethane anesthesia with the isoprenaline chronotropic effect 'in vivo' cannot be explained by a direct interference of urethane with beta-adrenoceptors at cardiac level.     

Reduction of isoprenaline induced tachycardia in pregnant rats. Role of a serum factor]

The in vivo isoprenaline (50 microgram/kg) induced tachycardia (beta 1 adrenergic stimulant effect) was decreased in pregnant (20th day) Rats compared to non pregnant Rats. The in vitro positive chronotropic effect of isoprenaline (10 ng/ml) on cultured Rat heart cells was abolished by pregnant Rat serum whereas progesterone (0.10 to 5 microgram/ml) or oestradiol (0.1 to 25 microgram/ml) were ineffective. The decreased beta 1 adrenergic responsiveness in pregnant Rats could be related to a seric factor, different from these two hormones.     

An analysis of the effects of urethane on cardiovascular responsiveness to catecholamines in terms of its interference with Ca++ mobilization from both intra and extracellular pools

Summary Urethane (1×10^–2–1×10^–1 M) reduced, in a concentration-dependent manner, both intra and extracellular Ca⁺⁺ dependent noradrenaline-induced contractions of perfused rabbit ear artery as well as the tonic contractions produced by perfusion with high K⁺ solution. However, a quantitative analysis of the data indicated that for urethane concentrations similar to those found in plasma during anesthesia urethane antagonism is confined to noradrenaline-induced contractions which depend upon the mobilization of Ca⁺⁺ from intracellular storage sites. In KCl-contracted arteries, urethane enhanced the relaxant effects of isoprenaline.—Urethane reduced the amplitude of contractions of spontaneously beating guinea-pig right atrium at concentrations which have only a limited effect on frequency. In addition, it decreased in a concentration-dependent manner the amplitude of isoprenaline-activated electrically driven, and K⁺ depolarized guinea-pig right ventricular strips. Urethane had no effect on the chrono and inotropic actions of isoprenaline on cardiac preparations. In in vivo experiments the chronotropic response to low doses of isoprenaline was significantly higher in urethane-treated as compared to unanesthetized rats. The higher dose of isoprenaline tested produced a significant fall in systolic blood pressure in urethane-anesthetized rats. A significant correlation exists between the chronotropic response to isoprenaline and resting heart rate values in urethane-anesthetized rats. These results indicate that urethane, at concentrations similar to those found in plasma during anesthesia selectively interferes with mobilization of Ca⁺⁺ from intracellular storage sites. In addition, the interference of urethane anesthesia with the isoprenaline chronotropic effect in vivo cannot be explained by a direct interference of urethane with -adrenoceptors at cardiac level.     

Effect of isoprenaline on dopamine receptors in the rabbit isolated renal artery

Summary The relaxant effects of isoprenaline on rabbit isolated renal artery and aorta were compared. The results suggest that although isoprenaline acts on -adrenoceptors in the aorta it stimulates dopamine receptors in the renal artery.     

Inhibition of isoprenaline-induced increase in plasma renin concentration by vasoconstrictors.

The vasoconstrictors angiotensin II, vasopressin and the alpha-sympathomimetic phenylephrine significantly inhibit the renin release caused by the beta-sympathomimetic isoprenaline. The mechanism of the inhibition is discussed.     

Sympathetic, muscarinic vasodilation in cranial vessels of the cat

Pressor responses evoked by stimulation of the preganglionic sympathetic trunk of the feline superior cervical ganglion have been recorded in vivo from the vascular bed perfused by one external carotid and the vertebral artery. When vasoconstrictor activity is blocked and potential vasodilator activity enhanced by close, intracarotid injection of guanethidine and prostaglandin F2 alpha respectively, stimulation evokes a weak pressor response followed, on cessation of stimulation, by a prolonged vasodilation lasting for 6-8 min. The magnitude and duration of the poststimulation vasodilation was reduced significantly by atropine. Due to the prolonged nature of the vasodilation, it is unlikely that a sympathetic cholinergic vasodilation in the classical sense is involved.     

VIP receptors and control of short circuit current in the human intestinal clonal cell line Cl.19A

At the maximally effective concentration of 10 nM, VIP induced a marked (12.5-fold stimulation above basal), and sustained increase in short circuit current in the human intestinal epithelial cell line Cl.19A grown on permeable filters and placed in Ussing chambers. Half-maximal increase of Isc was observed for 0.1 nM VIP. This was well correlated with the VIP-stimulated adenylate cyclase activity (ED50:0.07 nM). Binding studies using 125I-VIP indicated that Cl.19A cells express a peptide-specific VIP receptor with a dissociation constant of 0.07 nM. Covalent labeling of receptors followed by SDS-PAGE analysis of membrane proteins resulted in the identification of a 63,000 dalton binding protein in Cl.19A cells.     

Effects of several preoperative medications on fat cell lipolysis, and activity of adipose tissue cyclic AMP phosphodiesterase.

Effects were examined of atropine, diazepam, pethidene, promethazine, scopolamine, omnopon and papaverine on basal and noradrenaline-stimulated lipolysis in rat isolated fat cells and on rat adipose tissue cyclic AMP phosphodiesterase activity. Papaverine at high concentration (1 mM) inhibited both basal and hormone-stimulated lipolysis, whereas diazepam enhanced basal lipolysis. At a 'clinical dose', omnopon increased both basal and noradrenaline-stimulated lipolysis. Adipose tissue cAMP phosphodiesterase activity was strongly inhibited by 1 mM diazepam, papaverine, promethazine and omnopon (280 microgram ml-1). Lack of enhancement of lipolysis by the established cAMP phosphodiesterase antagonist papaverine, is compatible with simultaneous inhibition also of adipose adenyl cyclase. Diazepam-stimulated lipolysis is compatible with its phosphodiesterase inhibitory activity. It is proposed that papaverine-containing omnopon may offer some survival advantages during surgical stress by facilitating a caloric supply.     

Host defense against infections and inflammations: role of the multifunctional IL-6/IFN-beta 2 cytokine

IL-6/IFN-beta 2 appears to be one of the important mediators of the response to viral and bacterial infections and to shock. The biological effects now associated with IL-6/IFN-beta 2 include: stimulation of immunoglobulin secretion by mature B lymphocytes (BSF-2 activity), growth stimulation of plasmacytomas and hybridomas (HGF activity), activation of T cells, stimulation of hepatic acute phase protein synthesis (HSF activity), stimulation of hematopoiesis, cell differentiation (DIF activity), inhibition of tumor cell growth (AP activity) and other IFN-like effects. As a typical cytokine, IL-6/IFN-beta 2 is secreted by many cell types and acts in various combinations with other interleukins and interferons.     

Alpha-receptor subsensitivity of isolated atria from rats following repeated injections of phenylephrine or isoprenaline

Summary Daily injections of phenylephrine or isoprenaline for 4 or 7 days lowered the chronotropic sensitivity of -adrenoreceptors in isolated rat atria.This study was supported by grants from the National Science Council of Finland.     

Effects of receptor agonists on polyamine concentrations and spermidine/spermine N1-acetyltransferase activity in rat parotid gland

The polyamine putrescine might be formed via a degradation (catalyzed by spermidine/spermine N¹-acetyltransferase, SSAT) of the higher polyamines spermidine and spermine to putrescine. The involvement of different intracellular signal pathways in the regulation of putrescine formation was studied in explants and in cultured cells of rat parotid glands by using receptor agonists that activate separate second messenger systems, and measuring their effects on the concentrations of putrescine, spermidine and spermine and on the SSAT activity. The -adrenoceptor agonist isoprenaline, which is an activator of cAMP formation, increased the putrescine concentration and stimulated the SSAT activity. Pilocarpine, a drug that activates the muscarinic receptors and thereby enhances the phosphoinositide turnover, had no effect on either the polyamine concentrations or on the SSAT activity. Epidermal growth factor (EGF), which induces activation of a protein tyrosine kinase, had no effect on the polyamine concentrations or on the SSAT activity. The adenylate cyclase activator forskolin increased the glandular levels of putrescine. Taken together, these findings suggest that increases in putrescine concentration in cultured rat parotid gland cells are accompanied by accumulation of cAMP.     

Fluid fluxes in the ferret trachea

Summary A net reabsorption of fluid was observed in isolated ferret trachea under control conditions. Cholinergic stimulation resulted in net secretion of fluid while atropine blocked this response without any effect on the basal process of fluid reabsorption.Supported by research grants from the Cystic Fibrosis Foundation and the University of Florida (GML) and by a Pediatric Pulmonary Center award (A 2018) from the Division of Maternal and Child Health of the Department of Health and Human Services and Whitehall Foundation grant (No. 78-156 ck-1).     

Sympathetic,muscarinic vasodilation in cranial vessels of the cat

Summary Pressor responses evoked by stimulation of the preganglionic sympathetic trunk of the feline superior cervical ganglion have been recorded in vivo from the vascular bed perfused by one external carotid and the vertebral artery. When vasoconstrictor activity is blocked and potential vasodilator activity enhanced by close, intracarotid injection of guanethidine and prostaglandin F₂ respectively, stimulation evokes a weak pressor response followed, on cessation of stimulation, by a prolonged vasodilation lasting for 6–8 min. The magnitude and duration of the poststimulation vasodilation was reduced significantly by atropine. Due to the prolonged nature of the vasodilation, it is unlikely that a sympathetic cholinergic vasodilation in the classical sense is involved.     

Quantitative changes in beta-adrenergic responses of isolated atria from hyper- and hypothyroid rats

Concentration-response curves for the chronotropic and inotropic effects of isoprenaline, in the absence and presence of propranolol, were obtained on heart atria isolated from normo- or dysthyroid rats. Hyperthyroidism increased the chronotropic potency and efficacy of the beta-adrenergic agonist. The results are compatible with the view that thyroid hormone increases the density of functional beta-adrenoceptors in cardiac pacemaker tissue.     

Activité gonadotrope d'un extrait d'urines de femmes en ménopause

Summary An extract from pooled postmenopausal urine has been investigated for its gonadotrophic activity. Dose-response curves have been constructed for its total gonadotrophic activity (stimulation of estrogen secretion as reflected by the uterine weight increase in immature mice) and for its ICSH activity (stimulation of androgen secretion as reflected by the ventral prostate weight increase in hypophysectomised immature rats). In hypophysectomised immature rats, the extract produced repair of the interstitial cells of the testes and complete spermatogenesis. It contains, therefore, FSH as well as ICSH activity in comparable amounts, a fact which opens up interesting therapeutic possibilities.     

Molecular and structural effects of inverse agonistic mutations on signaling of the thyrotropin receptor – a basally active GPCR

Several mutations that decrease the basal signaling activity of G-protein coupled receptors (GPCRs) with pathogenic implications are known. Here we study the molecular mechanisms responsible for this phenotype and investigate how basal and further activated receptor conformations are interrelated. In the basally active thyroid stimulating hormone receptor (TSHR) we combined spatially-distant mutations with opposing effects on basal activity in double-mutations and characterized mutant basal and TSH induced signaling. Mutations lowering basal activity always have a suppressive influence on TSH induced signaling and on constitutively activating mutations (CAMs). Our results suggest that the conformation of a basally ‘silenced’ GPCR might impair its intrinsic capacity for signaling compared to the wild-type. Striking differences in conformation and intramolecular interactions between TSHR models built using the crystal structures of inactive rhodopsin and partially active opsin help illuminate the molecular details underlying mutations decreasing basal activity. G. Kleinau, H. Jaeschke: These two authors contributed equally to this work. Received 31 July 2008; received after revision 12 September 2008; accepted 19 September 2008     

Isoprenaline-like effects of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine on mechanical, biochemical and electrophysiological parameters in the mammalian heart

The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) mimicked the effects of isoprenaline on the force of contraction, the cAMP content and the slow Ca++ inward current (Isi) in isolated guinea pig papillary muscles. The results support the hypothesis that phosphodiesterase inhibitors and beta-adrenoceptor agonists exert their positive inotropic effects by increasing Isi via the common mediator cAMP.