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C Reithmann

Publications and source records attributed to C Reithmann.

51 records · Page 3Linked to original sources
PubMed1990

Stimulation and inhibition of adenylyl cyclase.

The adenylyl cyclase serves as a signal transduction system located in the plasma membrane of apparently every cell type of higher organisms. It generates the second messenger cyclic AMP, which plays an important regulatory role in many diverse biological processes such as glycogenolysis and lipolysis, cardiac inotropy and chronotropy, vascular tone, neurotransmitter and hormone release in neuronal and endocrine cells, respectively, and cell growth. Cyclic AMP acts by activating cyclic AMP-dependent protein kinases which phosphorylate various ubiquitous and cell-specific proteins, which covalent modifications cause the final cellular responses. The complete adenylyl cyclase signal transduction system is a multi-component system consisting of various receptors which recognize the extracellular signals, two guanine nucleotide-binding regulatory proteins (G-proteins) which transduce the signals from the receptors to the adenylyl cyclase and the effector enzyme which catalyzes the formation of cyclic AMP from ATP. This brief review focusses on our current knowledge about structure and function of the components involved in the stimulation and inhibition of adenylyl cyclase and provides some recent data on possible modifications of these components.

Adenylyl Cyclases↗
PubMed1989

Intrinsic sympathomimetic activity of beta-adrenoceptor antagonists: down-regulation of cardiac beta 1- and beta 2-adrenoceptors.

Prolonged treatment of cultured rat heart muscle cells containing beta 1- and non-muscle cells containing beta 2-adrenoceptors with beta-adrenoceptor antagonists devoid of intrinsic sympathomimetic activity had no effect on beta-adrenoceptor density. In contrast, antagonists with intrinsic sympathomimetic activity decreased beta-adrenoceptor density and response (adenylate cyclase stimulation) in both heart muscle (beta 1) and non-muscle cells (beta 2) by a maximum of about 50%. An even larger down-regulation of beta-adrenoceptors and loss of receptor-stimulated adenylate cyclase activity was induced by the full endogenous agonist, noradrenaline, with the beta-adrenoceptors of heart muscle cells (beta 1) being much more sensitive to the beta 1-selective noradrenaline than the heart non-muscle cell beta 2-adrenoceptors. When combined with noradrenaline, the antagonists with intrinsic sympathomimetic activity prevented the action of noradrenaline at both beta 1- and beta 2-adrenoceptors, thereby leading to an apparent up-regulation of receptor density and response. This apparent reversal from an agonist to an antagonist action was observed at much lower concentrations of noradrenaline at beta 1- than at beta 2-adrenoceptors. The data presented indicate that the beta-adrenoceptor antagonists with intrinsic sympathomimetic activity, but not those without, upon prolonged treatment decrease the density and responsiveness of both beta 1- and beta 2-adrenoceptors in cultured rat heart cells. This suggests that the intrinsic sympathomimetic activity of these agents is not a subtype-selective component. Furthermore, the agonist and antagonist activity of these agents apparently depends on the concomitant presence of an endogenous full agonist and an its own affinity and that of the partial agonist for the beta-adrenoceptor subtype.

Adenylyl Cyclases↗
PubMed1989

Mechanism of noradrenaline-induced heterologous desensitization of adenylate cyclase stimulation in rat heart muscle cells: increase in the level of inhibitory G-protein alpha-subunits.

The mechanism of heterologous desensitization of adenylate cyclase stimulation was studied in cultured neonatal rat heart muscle cells. After culturing of the cells for 3 days in the presence of 1 microM noradrenaline there was in addition to a 52% decrease in isoproterenol-stimulated adenylate cyclase activity, a lessening of the stimulation of beta-adrenoceptor-independent adenylate cyclase by guanosine-5'-O-(thiotriphosphate) and forskolin by 24 and 34%, respectively. The decrease in receptor-independent adenylate cyclase stimulation by forskolin, but not the attenuation of isoproterenol-stimulated adenylate cyclase activity, was abolished by pertussis toxin (PTX) pretreatment of the cells. Gi, the inhibitory G-protein of adenylate cyclase was therefore quantitated. Labelling of the Mr approximately 40 kDa PTX substrates in membranes of noradrenaline-treated cells was increased by 70% as shown by pertussis toxin-catalyzed ADP ribosylation of heart cell membranes. This increase was also seen in the presence of an excess of purified beta gamma-subunits of transducin and of GTP, suggesting that the increased labelling was not due to elevation of the level of beta gamma-subunits or increase in the concentration of GTP in the membranes of noradrenaline-treated cells. Analysis of the PTX substrates on high resolution urea/SDS-polyacrylamide gels revealed that at least two distinct PTX substrates (40 and 41 kDa) were present in rat heart cell membranes. The labelling of both substrates was increased in membranes of desensitized cells. Immunoblotting of heart cell membranes with anti-Gi alpha-antibodies demonstrated a marked increase in the amount of Gi alpha in membranes of noradrenaline-treated cells. In contrast, immunoblotting with anti-beta-antibodies showed that the level of the beta-subunit of G-proteins (36 kDa) was unchanged after noradrenaline exposure. The data indicate that prolonged treatment of rat heart muscle cells with noradrenaline leads to an increase in the level of alpha-subunits of Gi-proteins. This suggests that this increase is responsible for the observed heterologous desensitization of adenylate cyclase stimulation.

Adenosine Diphosphate↗
PubMed1988

Homologous vs. heterologous desensitization of the adenylate cyclase system in heart cells.

Exposure of cultured heart muscle cells to noradrenaline led to a decrease in the effects of isoproterenol and prostaglandin E1 on cAMP formation and contraction velocity. However, heterologous desensitization, as measured by prostaglandin E1 stimulation, only occurred at higher noradrenaline concentrations than homologous desensitization (isoproterenol stimulation). As the defects of the adenylate cyclase system in heart failure are attributed to noradrenaline-induced desensitization, it is concluded from the results that, in comparison to the subsensitivity to beta-adrenoceptor agonists in failing human hearts, a decrease in the responsiveness to other receptor-dependent adenylate cyclase stimulators should also occur but only at higher degrees of heart failure.

Adenylyl Cyclases↗
PubMed1988

Studies on the role of central catecholaminergic mechanisms in the antidotal effect of the oxime HI 6 in soman poisoned mice.

The effects of atropine and the oxime HI 6 on running performance, brain and plasma cholinesterase activity and brain catecholamines were investigated in mice intoxicated with sublethal doses of soman (100 micrograms/kg s.c.). The running time on a rotating mash wire drum (total running time 60 min) after injection of soman was reduced to 17.2 min. Treatment with atropine (10 mg/kg i.p.) or HI 6 (55 mg/kg i.p.) improved the running performance to 48.2 and 44.8 min, respectively. Cholinesterase activity was decreased in soman poisoned mice to 47.3% in plasma and 43.5% in brain. Therapy with the oxime HI 6 resulted in a reactivation of soman-inhibited peripheral cholinesterase to 76.6%, but failed to reactivate central cholinesterase. Dopamine levels in mice brain were elevated in soman poisoning by 23.2%, whereas noradrenaline levels remained unchanged. The increase in brain dopamine levels was antagonized by atropine as well as by HI 6. The results of this study lead to the speculation that central dopaminergic mechanisms may be involved in soman toxicity as well as in the antidotal action of atropine and the mainly peripherally acting oxime HI 6.

Animals↗
PubMed1987

The role of endogenous noradrenaline in the beta-blocker withdrawal phenomenon--studies with cultured heart cells.

An in vitro model to evaluate the role of endogenous noradrenaline in the beta-blocker withdrawal phenomenon is described: Beating chicken heart muscle cells (5000 beta 1-adrenoceptors/cell) and heart nonmuscle cells (3000 beta 2-adrenoceptors/cell) were cultured in serum-free, hormone-supplemented medium. Basal state, subtype selective down-regulation of beta-adrenoceptors by endogenous noradrenaline (decrease in receptor number, beta 1 more than beta 2) was simulated by addition of noradrenaline to the culture medium; chronic beta-blockade was simulated by exposure of the cells for 3 days to various beta-blockers (propranolol, no ISA; timolol, slight ISA; pindolol, strong ISA). Beta-blocker withdrawal phenomenon--increased response in isoproterenol-induced cAMP production and positive inotropy--is correlated with the increase in the number of beta-adrenoceptors after withdrawal of the drugs. Propranolol induces a withdrawal phenomenon at every degree of noradrenaline-induced basal state down-regulation of beta-adrenoceptors; in contrast, a withdrawal phenomenon by pindolol is only seen at a higher degree of beta-adrenoceptor down-regulation. In the presence of physiological noradrenaline concentrations pindolol affects beta-adrenoceptor subtypes in a qualitatively different manner: the number of beta 1-adrenoceptors is increased, the number of beta 2-adrenoceptors is decreased. This finding demonstrates that the intrinsic sympathomimetic activity of nonselective beta-blockers can manifest itself only if the receptors are not strongly down-regulated. As beta 2-adrenoceptors are present in a much less down-regulated state than beta 1, ISA mainly acts on beta 2-adrenoceptor subtype, thus, presenting a beta 2-"pseudo-selectivity" of ISA.

Adrenergic beta-Antagonists↗
PubMed1987

Endogenous noradrenaline and beta-adrenoceptor regulation in cultured heart cells.

a) Cultured heart muscle cells from chicken embryos and neonatal rats possess about 5000 beta 1-adrenoceptors/cell, heart non muscle cells from these species about 3000 beta 2-adrenoceptors/cell. Agonist binding in heart muscle cells results in cAMP formation and in a positive inotropic effect. b) Beta 1- and beta 2-adrenoceptors of heart cells are downregulated to different degrees in the presence of physiological noradrenaline concentrations in the culture medium. c) Using chicken heart cells in culture, a cellular model of the beta-blocker withdrawal phenomenon in the heart has been established, elucidating the modifying action of endogenous catecholamines and the role of ISA (partial agonist activity) in this phenomenon and classifying the so-called "beta 2-pseudo-selectivity" of the ISA action. d) In the absence of added catecholamines, (-)propranolol exposure of the cells for 3 days increases the number of beta-adrenoceptors of chicken heart muscle cells up to 170% of control, while in rat heart muscle cells the receptor number remains unchanged. The mechanism of this "basal" receptor downregulation in chicken heart muscle cells is unclear. The data presented demonstrate the usefulness of heart cell cultures in the study of beta-adrenoceptor regulation of the heart.

Adenylyl Cyclases↗
PubMed1985

Cardiac glycoside tolerance in cultured chicken heart muscle cells--a dose-dependent phenomenon.

In cultured heart muscle cells from 10-13 day-old chicken embryos, the effects of acute (4 h) and chronic (3 days) exposure of the cells to varying concentrations of ouabain have been studied. In these cells, the cardiac glycoside ouabain binds to a specific cardiac glycoside receptor (KD = 4 X 10(-7) M; 750,000 receptors/cell). Binding to this receptor results in inhibition of active Na+/K+-transport [EC50 for active (86Rb+ + K+)-influx = 4 X 10(-6) M], and in an increase in beating velocity ("positive inotropic effect"; EC50 = 4 X 10(-7) M); toxic signs (arrhythmias) appear at concentrations greater than or equal to 6 X 10(-7) M. During exposure of the cells to 3 X 10(-6) M ouabain for 3 days, tolerance develops with respect to both the positive inotropic and the toxic effect. The mechanism underlying this tolerance is identified as an increase in the number of active sodium pump molecules per cell, while the binding properties of the cardiac glycoside receptor remain unchanged. The development of cardiac glycoside tolerance is only observed in the presence of severe impairment of Na+/K+-homeostasis, due to cardiac glycoside-induced inhibition of active Na+/K+-transport. This, however, only occurs in the presence of toxic (receptor occupation greater than or equal to 60%), but not in the presence of positive inotropic, non-toxic (receptor occupation 20-60%), ouabain concentrations. We conclude that the development of cardiac glycoside tolerance during long-term treatment in patients with heart failure should not occur with submaximal dose regimens, when toxic signs (arrhythmias) are absent.

Animals↗
PubMedDate not supplied

Noradrenaline-induced desensitization in cultured heart cells as a model for the defects of the adenylate cyclase system in severe heart failure.

Cultivation of rat heart muscle cells for up to 5 days in the presence of 10(-6) mol/l (-)noradrenaline leads to a desensitization of the cells to inotropic stimulation and cAMP formation at different levels of the cAMP-system: The decrease in the responsiveness to the beta-adrenoceptor agonist isoprenaline quantitatively parallels the down-regulation of beta 1-adrenoceptors. The impairment of the positive inotropic effect of the phosphodiesterase-inhibitor IBMX is associated with a diminished basal cAMP-formation but an unchanged phosphodiesterase-activity in noradrenaline-treated cells. This decrease in basal cAMP-accumulation as well as the attenuation of the forskolin-stimulated cAMP-formation indicate that a defect in the adenylate cyclase system beyond the receptor must be involved in noradrenaline-induced desensitization. In contrast to the diminished effectiveness of cAMP-increasing agents the positive inotropic effect of ouabain and the alpha-adrenoceptor-mediated positive inotropic effect of phenylephrine are unaltered in noradrenaline-treated cells. It is concluded from these results that in addition to the down-regulation of beta 1-adrenoceptors a defect on the post-receptor level of adenylate cyclase occurs in noradrenaline-induced desensitization not affecting mechanisms beyond the catalytic subunit of adenylate cyclase. As these findings are very similar to those observed in isolated preparations from severely failing human hearts, the presumption is confirmed that noradrenaline-induced desensitization might constitute an important etiological factor in the subsensitivity of failing human hearts to inotropic stimulation.

1-Methyl-3-isobutylxanthine↗
PubMedDate not supplied

Mechanisms in acute septic cardiomyopathy: evidence from isolated myocytes.

Although often not considered, the heart is one of the targets of multiple organ failure in sepsis and septic shock, with myocardial depression being a prominent component of this "acute septic cardiomyopathy". Hypotheses concerning the etiology of this depression are increasingly elucidated on a cellular level, including dysfunction of the beta-adrenoceptor/G protein/adenylate cyclase system, calcium channel blockade by cardiodepressant factor, contractile impairment by activated leucocytes, as well as inhibition of protein synthesis by Pseudomonas exotoxin A. In the search for "mechanisms of myocardial depression in sepsis", isolated cardiomyocytes may play a role as research tools with respect to: a) discrimination between direct and indirect cardiodepressant effects; b) identifying not only the acute, but also chronic toxin- and mediator-induced cardiodepression; c) clarification of the mechanism of action of cardiodepressant bacterial toxins and sepsis mediators; d) establishment of in vitro models of leucocyte-mediated cardiodepression in sepsis.

Acute Disease↗