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J C Rüegg

Publications and source records attributed to J C Rüegg.

At least 37 records · Page 2Linked to original sources
PubMed1995

Myofibrillar Ca2+ sensitivity of cardiomyopathic hamster hearts.

We studied the Ca2+ responsiveness of skinned muscle fibre preparations from the right and left ventricles of normal (FIB) and genetically cardiomyopathic (Bio-To-2) Syrian hamsters. Thus, we compared the Ca2+/force relationships of preparations from myopathic hamsters to those of age-matched (11-16 months old) normal animals. The pCa (i.e. -log10 [Ca2+]) required for 50% force activation (Ca2+ sensitivity) was higher in the myopathic hamsters than in controls (pCa50 values of 5.3 +/- 0.03 and 5.17 +/- 0.04, respectively); this difference might be due to an alteration in regulatory proteins. Indeed, after extraction (with vanadate) and replacement of troponin I with bovine cardiac troponin the pCa50 values were similar (pCa 5.35) to those of bovine ventricular fibres. The Ca2+ sensitizer EMD 53998 (10 microM) increased Ca2+ sensitivity in preparations from normal and cardiomyopathic hamsters equally, by 0.4 pCa units. Incubation of fibre bundles with the catalytic subunit of cyclic-adenosine-monophosphate-dependent protein kinase decreased Ca2+ sensitivity, thereby "normalizing" the enhanced Ca2+ responsiveness of fibres from cardiomyopathic hamsters. It is not clear, however, whether the pathologically increased Ca2+ sensitivity of the hearts of aged myopathic hamsters reflects a maladaptation, or a compensatory mechanism of the failing heart.

Actin Cytoskeleton↗
PubMed1995

Effects of troponin-I extraction with vanadate and of the calcium sensitizer EMD 53998 on the rate of force generation in skinned cardiac muscle.

The rate of tension development following release of ATP from caged-ATP in the presence of calcium was studied in skinned cardiac fibres from swine. A low-force rigor state was obtained by using butanedione monoxime (BDM) during the induction of rigor. BDM was washed out and following release of ATP in the presence of Ca2+ (pCa 4.3), the muscles contracted with an apparent rate of about 2 s-1 at 22 degrees C. After treatment with 10 mM vanadate to extract troponins I and C the fibres contracted independently of calcium. The rate of contraction upon release of ATP was slower than prior to extraction and was independent of [Ca2+]. Since treatment with vanadate has been shown to extract about 90% of troponin-I the results suggest that the muscles under these conditions are partially activated by removal of an inhibition of cross-bridge interaction by troponin I. A partial recovery of force was obtained by prolonged incubation in DTT containing solutions possibly reflecting reconstitution with troponin I still present in the fibre bundle. Treatment with a solution containing whole troponin caused almost complete recovery of calcium sensitivity and rate of force development. The calcium sensitizer EMD 53998 increased rates of contraction in a dose dependent manner, suggesting that this compound increases force and calcium sensitivity by increasing the cross-bridge attachment rates.

Adenosine Triphosphate↗
PubMed1994

The calcium sensitizer EMD 53998 antagonizes phosphate-induced increases in energy cost of isometric tension in cardiac skinned fibres.

We have investigated whether a Ca(2+)-sensitizing substance, the thiadiazinone derivative EMD 53998, can alter the ratio of ATPase activity to force, i.e. the tension cost in skinned fibres of swine cardiac trabecula in which the tension cost was increased by inorganic phosphate. In the presence of 10 mM inorganic phosphate (Pi) and thapsigargin 20 microM, EMD 53998 reduced the energy cost of isometric tension over the entire range of activating Ca2+ concentrations, resulting in a consistent change in slope (approximately 20% decrease) of the ATPase/force relation. We confirmed that in the absence of added phosphate and at maximal Ca2+ activation EMD 53998 had little if any effect on tension cost. We had previously reported that the effects of EMD 53998 and Pi on calcium sensitivity and maximum isometric tension are mutually antagonistic and our new energy data now support the proposal that EMD 53998 functionally antagonizes the effects of Pi on crossbridges. The decrease in the slope of the relation between ATPase and force caused by EMD 53998 may be interpreted to reflect either a decrease in the rate of 'detachment' (g(app)) of crossbridges or an increase in average force per crossbridge, as predicted by classical crossbridge models. Since the Pi release step of the crossbridge cycle is associated with the rate of 'attachment' (f(app)) rather than g(app), we conclude that the decrease in tension cost with EMD 53998 most likely reflects an increased force per crossbridge.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Diphosphate↗
PubMed1994

Coordination of nuclear and mitochondrial gene expression during the development of cardiac hypertrophy in rats.

We studied the coordination of nuclear and mitochondrial gene expression during cardiac hypertrophy following aortic stenosis or thyroid hormone treatment in rats. We measured mRNA levels for representative subunits of cytochrome-c oxidase, two encoded by mitochondrial DNA and two encoded by the nucleus, as well as the levels of one mitochondrial rRNA. In both models of hypertrophy, an increase of total tissue RNA, reflecting mainly cytosolic ribosomes, accompanied the increase in ventricular weight. Relative levels of mitochondrial rRNA remained unchanged, indicating a net synthesis of mitochondrial ribosomes as well. In both models, cytochrome-c oxidase activity and nuclear-encoded mRNAs remained fairly constant, whereas levels of mitochondrial mRNAs were transiently decreased 24 h after the growth stimulus. We conclude that, in the initial phase of hypertrophy, the signal regulating the synthesis of mitochondrial rRNA is synchronized with nuclear gene expression, whereas the signal regulating mitochondrial mRNA synthesis is not. We postulate that differential regulation of mitochondrial transcription and premature termination of the polycistronic transcript (the latter giving rise to the mitochondrial rRNAs) account for the observed results.

Animals↗
PubMed1994

Infusion of oxidized glutathione enhances postischemic segment-shortening in dog hearts.

Oxidized glutathione (GSSG) but not its reduced form (GSH) is taken up by intact myocardial cells, and is rapidly converted into GSH. Reduced glutathione is an important intracellular defense against oxygen-derived free radicals and has been found to enhance calcium sensitivity in skinned cardiac fibers. We have investigated the effects of intravenous GSSG on left ventricular systolic pressure, maximal rate of rise of pressure and regional segment-shortening in dogs subjected to occlusion of the left anterior descending artery for 30 minutes, followed by 45 minutes reperfusion. Starting 10 minutes before reperfusion, the dogs were randomly treated with either GSSG (100 mM, 5 ml/min, n = 5) or Ringer's solution (5 ml/min, n = 5) until 30 minutes of reperfusion. Myocardial blood flow was measured by radioactive microspheres. Infusion of GSSG increased total glutathione content in both ischemic (47 +/- 16 mumol/g protein) and nonischemic myocardium (71 +/- 17 mumol/g protein) as compared to controls (23 +/- 2 mumol/g protein, p < 0.05). In both groups paradoxical wall motion occurred in the ischemic region during occlusion. On reperfusion, regional dyskinesia persisted in controls; while, in glutathione-treated dogs, systolic segment-shortening reached half the baseline values (p < 0.05, treated vs controls, at 15, 30, 45 minutes reperfusion). During ischemia the area of pressure-length loops, obtained from simultaneous recordings of left ventricular pressure and regional segment length, decreased to 30 +/- 7% of baseline in controls and to 40 +/- 18% of baseline in GSSG-treated animals. After 45 minutes reperfusion it was restored to 78 +/- 22% baseline in treated hearts but was still 36 +/- 16 of baseline in controls (p < 0.05). We conclude that infusion of GSSG increases the intracellular stores of glutathione and improves the contractile state of postischemic myocardium.

Animals↗
PubMed1994

Inotropic effect of the cardioprotective agent 2,3-butanedione monoxime in failing and nonfailing human myocardium.

The drug 2,3-butanedione monoxime (BDM) was suggested to be a potent cardioprotective agent useful for cardiopreservation. The present study investigated the activity of BDM and its mechanism of action in human myocardium. In electrically driven left ventricular papillary muscle strips and right atrial trabeculae from failing (heart transplants) and nonfailing (donor hearts) human myocardium, isometric force development and the force-frequency relationship were examined. To study the sarcolemmal actions of BDM, competition experiments with 125I-iodocyanopindolol, 3H-ouabain and 3H(+)PN 200-110 were performed. The effect of BDM on the contractile apparatus was tested by investigating its effects on Ca++ sensitivity and on the relaxation parameters of skinned fiber preparations. In papillary muscle strips and in atrial trabeculae, BDM (0.001-30 mM) depressed the isometric force of contraction in a concentration-dependent manner. BDM was more potent in atrial than in ventricular tissue and it shifted the Ca++ concentration-response curve in atrial and ventricular tissue to the right. The potency of BDM to depress force development was significantly lower compared with the L type of Ca++ channel antagonist nifedipine in both atrial and ventricular myocardium. In the presence of 10 mM BDM, the force-frequency relationship becomes positive in failing myocardium but not in the presence of 1 mM BDM, which did not affect the specific binding of 125I-iodocyanopindolol, 3H-ouabain or 3H(+)PN 200-110. This indicated there was no action on beta adrenoceptors, cardiac glycoside receptors and dihydropyridine-type Ca++ channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

A synthetic peptide mimics troponin I function in the calcium-dependent regulation of muscle contraction.

A new technique for treating skinned cardiac muscle fibers has been developed in which troponin I is extracted, giving rise to unregulated fibers. The effect of the 12-residue troponin I peptide on these fibers indicates that this region of troponin I is solely responsible for muscle relaxation (inhibition of force). Furthermore, troponin I peptide-troponin C reconstituted fibers are stable through several contraction-relaxation cycles indicating the peptide can switch binding sites between actin and troponin C. The troponin I peptide can substitute for the native protein as part of the calcium-sensitive molecular switch that controls muscle regulation.

Amino Acid Sequence↗
PubMed1993

Effect of myosin heavy chain peptides on contractile activation of skinned cardiac muscle fibres.

Peptides derived from the sequence of the S1 domain of the myosin heavy chain were tested for their effects on the regulation of cardiac contractility. Basal calcium responsiveness of the contractile apparatus in terms of isometric tension generation and ATPase was determined in chemically demembranated ventricular fibre bundles. Incubation with a series of peptides derived from the peptide sequence around SH thiol group (Cys 707) resulted in a measurable increase in isometric tension and ATPase activity at sub-maximal concentrations of calcium but not at saturating levels of calcium activity, thus demonstrating a "calcium-sensitizing" effect of these peptides. The effects of two of these peptides, S1 687-716 and S1 701-717, are demonstrated to mimic, but importantly were not additive with, the calcium sensitization induced by lowering ATP concentration to 10 microM from 10 mM. This suggests the possibility of a similar mechanism of action underlying both types of sensitization. Because these effects demonstrate tissue specificity, were sensitive with respect to potency to not only amino acid composition but also sequence, and could not be duplicated by a similarly charged, non-homologous peptide, we attribute the effects to be specific to the sequences of these peptides. These data provide further evidence that the sequence between residues 687 and 717 of the S1 domain of the myosin heavy chain influences the calcium responsiveness of the contractile apparatus.

Actins↗
PubMed1993

Contractile properties of skinned preparations from ischaemic canine myocardium and coronary arteries.

The influence of prolonged ischaemia on the regulation of contraction in the myocardium and in the smooth muscle of coronary arteries was investigated. Chemically skinned preparations were used which enabled the contraction to be studied with the environment of the contractile filaments controlled. Myocardial ischaemia was produced in anesthetized adult beagle dogs by occlusion of the left anterior descending artery for 3 h and followed by 30 min reperfusion. Myocardial tissue and segments from coronary arteries were obtained from the ischaemic infarcted wall region ("in vivo ischaemic") and compared with control preparations from perfused coronary arteries and from the free wall of the left ventricle. Coronary and myocardial preparations were also obtained from the heart after a 3 h period in vitro under anoxic conditions at 37 degrees C ("in vitro ischaemic") simulating a state of extreme ischaemia. Control myocardial fibres were fully relaxed at pCa (-log-[Ca2+]) 9 and developed 24 +/- 5% (n = 7) of maximum force at intermediate calcium concentration (pCa 5.5). In contrast, the in vivo and in vitro ischaemic preparations produced force at pCa 9 (28 +/- 13 and 39 +/- 8%, respectively, n = 5 and 7) and showed an increased force development at pCa 5.5 (53 +/- 11 and 75 +/- 5%). The in vivo and in vitro ischaemic coronary arteries relaxed more slowly following calcium removal than control vessels. The in vitro ischaemic vascular preparations developed active force at pCa 9 and showed increased levels of myosin light chain phosphorylation and reduced phosphatase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1993

[Modulation of calcium sensitivity in the heart muscle--physiology, pathophysiology and pharmacology].

Angiotensin II has a plethora of different actions in the heart including, for instance, also slight positive inotropic effects. Thus cardiotonic therapy with angiotensin converting enzyme (ACE) inhibitors should possibly be complemented by positive inotropic drug therapy. With this respect, novel cardiotonic drugs that increase calcium sensitivity of myofilaments such as pimobendan (CAS 118428-36-7) might be particularly useful, as they will increase force with little, if any concomitant increase in myoplasmic free calcium. These drugs mimic mechanistically the Frank-Starling-Mechanism or alpha-adrenergic effects which also involve, at least partly, calcium sensitization of the myofilaments. The latter may therefore be a useful therapeutic principle, in particular in cases where myofibrillar calcium responsiveness is pathologically depressed such as for instance in the stunned myocardium or under hypoxic conditions.

Animals↗
PubMed1992

The positive inotropic calcium sensitizer EMD 53998 antagonizes phosphate action on cross-bridges in cardiac skinned fibers.

The diazinone derivative EMD 53998 sensitizes skinned myocardial fibers to Ca2+ and enhances maximal calcium-activated force (pCa = 4.5) by approximately 100%; the EC50 is 10 microM in the absence and about 30 microM in the presence of added inorganic phosphate (10 mM). Although concentrations of added phosphate as low as 0.5 mM inhibit force, at high concentrations of EMD 53998 (> or = 50 microM), phosphate only inhibits at concentrations exceeding 20 mM. These data suggest that the effects of EMD 53998 and phosphate are mutually antagonistic. Importantly, both EMD 53998 and phosphate had similar effects on force generation in troponin I-depleted (Ca(2+)-independent) skinned fibers, thus demonstrating that these compounds are likely to affect cross-bridges directly and not via the Ca(2+)-regulatory system.

Animals↗
PubMed1992

Troponin replacement in permeabilized cardiac muscle. Reversible extraction of troponin I by incubation with vanadate.

Calcium-dependent regulation of tension and ATPase activity in permeabilized porcine ventricular muscle was lost after incubation with 10 mM vanadate. After transfer from vanadate to a vanadate-free, low-Ca2+ solution (pCa greater than 8), the permeabilized muscle produced 84.8% +/- 20.1% (+/- S.D., n = 98) of the isometric force elicited by high Ca2+ (pCa approximately 4.5) prior to incubation with vanadate. Transfer back to a high Ca2+ solution elicited no additional force (83.2% +/- 18.7% of control force). SDS-PAGE and immunoblot analysis of fibers and solutions demonstrated substantial extraction (greater than 90%) of Troponin I (TnI). Calcium dependence was restored after incubation with solutions containing either whole cardiac troponin or a combination of TnI and troponin C subunits. This reversible extraction of troponin directly demonstrates the role of TnI in the regulation of striated muscle contractility and permits specific substitution of the native TnI with exogenously supplied protein.

Animals↗
PubMed1992

Counting target molecules by exponential polymerase chain reaction: copy number of mitochondrial DNA in rat tissues.

In this report, we show that the actual number of target molecules of the polymerase chain reaction can be determined by measuring the concentration of product accumulating in consecutive cycles. The equation describing product accumulation, log Nn = log eff x n + log N0, can be analyzed by linear regression and the molar concentration of target at cycle zero, N0, is obtained. Using this new approach, the actual content of mitochondrial DNA was determined in rat tissues and ranged from 116 x 10(9) molecules/g in fast-twitch skeletal muscle to 743 x 10(9) molecules/g in liver. Using morphometric data from the literature, mitochondria were found to contain 1 to 3 DNA molecules. There was no relation between the oxidative capacity of a tissue and its content of mitochondrial DNA, indicating that transcriptional and posttranscriptional mechanisms rather than gene dosage, as postulated by others, determine to what extent the mitochondrial genome is expressed.

Animals↗
PubMed1991

Peptide competition of actin activation of myosin-subfragment 1 ATPase by an amino terminal actin fragment.

The amino-terminal region of actin participates in the binding of myosin subfragment 1 (S1) during cross-bridge cycling, thereby assisting in the activation of the magnesium-dependent myosin ATPase. Effects of three actin fragments on the magnesium-dependent S1 and acto-S1 ATPase activities in solution were studied. One of the peptides, containing residues actin 1-44, mimicked the S1 ATPase-activating properties of actin and in turn inhibited acto-S1 ATPase both in a concentration-dependent manner. This suggests peptide competition for the actin binding site on myosin. The other fragments, residues actin 1-18 and 82-119, respectively, had no detectable effect on S1- and acto-S1 ATPase activity.

Actins↗
PubMed1991

What does TnCDANZ fluorescence reveal about the thin filament state?

Fluorescence of skinned psoas fibres reconstituted with the troponin C subunit labelled with the fluorescent probe dansylaziridine (TnCDANZ) increases upon activation with Ca2+. This fluorescence enhancement is due to Ca2+ binding to the Ca(2+)-specific binding sites of TnCDANZ and attachment of cross-bridges to the actin filament. We found that approximately 20% of the enhanced fluorescence signal derived from Ca2+ binding to TnCDANZ and 80% from cross-bridge attachment during maximal activation. Furthermore we studied the effects of different cross-bridge states on TnCDANZ fluorescence. "Weakly" bound, non-force-generating cross-bridge states (pCa 8, low ionic strength) and rigor cross-bridges revealed similar effects on the TnCDANZ fluorescence. "Strongly" attached, force-generating states, however, increased fluorescence to the greatest extent. These results suggests a complex system of reciprocal couplings between TnC and different attached cross-bridge states. Cooling or increase of inorganic phosphate decreased isometric force but hardly decreased fluorescence, suggesting the accumulation of attached cross-bridge states with low tension output.

Actin Cytoskeleton↗
PubMed1991

Contractile protein interactions in smooth muscle.

Smooth muscle tone and 'holding economy' depend on the rate constants governing the cross-bridge cycle. Thus, calcium activation via calmodulin-dependent myosin light chain phosphorylation may determine the apparent rate constant ('f') at which cross-bridges enter the force-generating state, forming actin-attached, strongly bound cross-bridges. This phosphorylation of the light chain may be inhibited in skinned fibers by a peptide mimic of the calmodulin recognition site of the myosin light chain kinase (RS 20) that relaxes smooth muscle. In smooth muscle, the apparent cross-bridge detachment rate constant ('g') also seems to be variable, a low constant allowing for a high holding economy and low shortening velocity in the 'latch state'. It may also account for force maintenance at low levels of myosin phosphorylation. Additionally, cross-bridge attachment may, however, be also controlled by other regulatory proteins such as calponin and caldesmon.

Actins↗