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1.
Int Immunopharmacol ; 19(2): 308-16, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24530918

ABSTRACT

Autoantibodies against ß1-adrenoceptors (ß1-ARs) have been detected in the serum of patients with various cardiac diseases; however, the pathological impact of these autoantibodies (ß1-AABs) has only been evaluated in cardiac tissue. The purpose of the present study was to evaluate whether ß1-AABs have deleterious effects on vascular reactivity in rats. An enzyme-linked immunosorbent assay was used to detect ß1-AABs in sera from immunized rats over a period of 1-3 months using the peptidic sequence of the second extracellular loop of human ß1-AR. Functional studies were performed in thoracic aortic (TA) and small mesenteric artery (SMA) rings from immunized rats. Following pre-contraction with phenylephrine (0.3 µM and 3 µM for the TA and SMA respectively), cumulative concentration-response curves (CCRCs) to various ß-AR agonists (isoproterenol, dobutamine, salbutamol, SR 58611A), acetylcholine, A23187, and sodium nitroprusside (SNP) were then plotted. The relaxations induced by dobutamine, SR 58611A, and acetylcholine were significantly impaired, but salbutamol-induced relaxations were not affected, in both vessels from immunized rats. A significant impairment of isoproterenol-induced relaxation was only observed in SMA. CCRCs to SNP were not modified in either of the vessels. A23187-induced relaxation was impaired in immunized rats. Following pretreatment with L-arginine, vasorelaxation to acetylcholine and SR 58611A was restored in immunized rats. This study demonstrates that immunization against the second extracellular loop of ß1-ARs has a deleterious impact on vasorelaxations in the TA and SMA of rats, involving alterations in endothelium-dependent NO signaling pathways.


Subject(s)
Antibodies/pharmacology , Aorta, Thoracic/drug effects , Mesenteric Arteries/drug effects , Peptides/pharmacology , Receptors, Adrenergic, beta-1/immunology , Adrenergic beta-Antagonists/pharmacology , Animals , Aorta, Thoracic/physiology , Endothelium, Vascular/drug effects , Immunization , Immunoglobulin G/immunology , Male , Mesenteric Arteries/physiology , Protein Conformation , Rats , Rats, Wistar , Receptors, Adrenergic, beta-1/chemistry , Vasodilation/drug effects
2.
Ann Cardiol Angeiol (Paris) ; 61(3): 167-72, 2012 Jun.
Article in French | MEDLINE | ID: mdl-22633471

ABSTRACT

PURPOSE: To evaluate the effect of active immunization with a peptide corresponding to the second extracellular loop of the human beta1-adrenoceptors (ß(1)-AR) on the reactivity of Wistar rat isolated aorta. METHODS: Nine-week-old Wistar rats were actively immunized for 3months with a peptide corresponding to the second extracellular loop of the human ß(1)-AR. Specific immunoglobulins G (IgG) were characterized by Elisa and the bicinchonic acid protein assay and their functionality were tested in isolated ventricular cardiomyocytes (IVC) from control rats. Aortic rings isolated from control or immunized rats were mounted in organ baths. Then, contractile curves to phenylephrine (1nM to 300µM) and relaxant curves to acetylcholine (1nM to 100µM) and isoprenaline (1nM to 30µM) were established. RESULTS: Cell shortening increased dose-dependently in rat IVC superfused with IgG containing ß(1)-AR antibodies (10 or 25µg/mL). Isoprenaline-induced positive inotropy was strongly reduced in IgG containing ß(1)-AR antibodies preincubated (3h) IVC. Phenylephrine-and acetylcholine-induced aortic responses were greatly inhibited in immunized rats compared to control ones. However, active immunization did not influence the isoprenaline-mediated relaxation. CONCLUSIONS: The present work confirms that ß(1)-AR antibodies directed against the second extracellular loop of ß(1)-AR induce a positive inotropic effect in adult rat IVC. Moreover, we demonstrated, for the first time, that 3-month immunization with ß(1)-AR peptide was associated with altered aortic endothelial function without change in the ß-AR-mediated vasorelaxation.


Subject(s)
Aorta , Autoantibodies/immunology , Endothelium, Vascular , Immunoglobulin G/blood , Receptors, Adrenergic, beta-1/immunology , Acetylcholine/pharmacology , Adrenergic beta-Agonists/pharmacology , Algorithms , Animals , Aorta/drug effects , Computer Graphics , Disease Models, Animal , Endothelium, Vascular/drug effects , Humans , In Vitro Techniques , Isoproterenol/pharmacology , Male , Myocytes, Cardiac/drug effects , Phenylephrine/pharmacology , Rats , Rats, Wistar , Vasoconstrictor Agents/pharmacology , Vasodilator Agents/pharmacology
3.
Vet J ; 192(3): 368-73, 2012 Jun.
Article in English | MEDLINE | ID: mdl-21986319

ABSTRACT

The direct vasodilatory action of pentoxifylline (1-(5-oxohexyl)-3,7-dimethylxanthine) and its signalling pathway was evaluated in equine digital veins. Cumulative concentration-response curves to pentoxifylline (1 nM to 300 µM) were recorded in phenylephrine-precontracted equine digital vein rings under different experimental conditions. Relaxation to pentoxifylline was partially inhibited by endothelium removal, but was unaltered by CGS-15943 (a non-xanthine adenosine receptor antagonist; 3 µM). Nitric oxide synthase (NOS), soluble guanylate cyclase and cyclooxygenase (COX) inhibitors (Nω-nitro-L-arginine methyl ester (100 µM), ODQ (30 µM) and indomethacin (10 µM), respectively) significantly reduced the maximum relaxation induced by pentoxifylline. Moreover, pentoxifylline-induced relaxation was strongly reduced by Rp-8-Br-PET-cyclic guanosine monophosphate-S (a protein kinase G inhibitor; 3 µM), but remained unaffected by H-89 (a protein kinase A inhibitor; 2 µM). Pentoxifylline-induced relaxation was associated with a 3.4-fold increase in tissue cGMP content. To investigate whether pentoxifylline can affect cAMP- and cGMP-mediated relaxations, curves to forskolin, to sodium nitroprusside (SNP) and 8-bromo-cGMP were also recorded in endothelium-denuded equine digital vein rings pretreated with pentoxifylline (10 and 100 µM). Pentoxifylline only potentiated the SNP-mediated relaxation at the highest concentration (100 µM). Thus, pentoxifylline relaxed equine digital veins via endothelium-dependent and endothelium-independent components. The effect was mediated through both the NOS and COX pathways and could also result from inhibition of cGMP specific-phosphodiesterase activity at the highest concentrations used.


Subject(s)
Horses/physiology , Pentoxifylline/pharmacology , Vasodilation/drug effects , Vasodilator Agents/pharmacology , Veins/drug effects , Animals , Colforsin/pharmacology , Cyclic GMP/analogs & derivatives , Cyclic GMP/pharmacology , Dose-Response Relationship, Drug , Endothelium, Vascular/physiology , Indomethacin/pharmacology , Nitroprusside/pharmacology , Tissue Culture Techniques
4.
Arch Mal Coeur Vaiss ; 96(9): 905-13, 2003 Sep.
Article in English | MEDLINE | ID: mdl-14571645

ABSTRACT

Knowledge of the sympathetic system is a basic element in the understanding of numerous physiological and physiopathological phenomena. In the two last decades, new pharmacological, biochemical and molecular tools have changed our approach to the roles of beta-adrenoceptors in the cardiovascular system. In the heart, the positive inotropic effect of predominant beta 1-adrenoceptor stimulation is classically recognised. Several studies reveal a significant physiological relevance of the beta 2-adrenoceptor which could activate different signalling pathways in addition to that of cAMP. Moreover, the detection of a third beta-adrenoceptor subtype, beta 3, in human heart, responsible for a negative inotropic effect through a NO signalling pathway, has changed the classically admitted paradigm on the regulation of heart function by the beta-adrenergic system. The identification of atypical beta-adrenoceptors, based on pharmacological tools, led to the discovery of "putative" beta 4-adrenoceptors, which constituted a low affinity state of the beta 1-adrenoceptors. In vessels, all beta-adrenoceptors subtypes, beta 1, beta 2 and beta 3, mediated a vasodilation, but the signalling pathway involved in this effect was variable according to their localization (endothelial or smooth muscle cells), the species and the vascular bed. beta-adrenoceptors are involved in several cardiovascular disease and could constitute a determinant therapeutic target. The efficiency of some beta-blockers used in the treatment of heart failure could result from action on beta 3-adrenoceptors. Moreover, a mutation of the beta-adrenoceptor subtype suggested a role in hypertension and diabetes mellitus.


Subject(s)
Cardiovascular Diseases/physiopathology , Cardiovascular Physiological Phenomena , Receptors, Adrenergic, beta-3/physiology , Adrenergic beta-Antagonists/pharmacology , Blood Vessels , Diabetes Mellitus/physiopathology , Heart , Humans , Hypertension/physiopathology , Signal Transduction , Vasodilation
5.
Cardiovasc Res ; 50(3): 486-94, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11376624

ABSTRACT

OBJECTIVE: Protein kinase C (PKC) is thought to be involved in the regulation of the mammalian cardiac excitation-contraction coupling process by vasoactive peptides like endothelin-1 (ET-1). However, the demonstration of a causal link between activation of specific PKC isoforms and the increase in contractility mediated by ET-1 is still inferential. METHODS: By means of adenovirus-mediated gene transfer, we specifically overexpressed PKC epsilon in cultured adult rabbit ventricular myocytes (Ad-PKC epsilon). Myocyte shortening and [Ca2+]i transients under basal and ET-1-stimulated conditions were measured in Ad-PKC epsilon and Ad-LacZ control transfected cells. RESULTS: Infection with Ad-PKC epsilon resulted in a strong, virus dose-dependent increase in PKC epsilon protein levels, whereas protein expression of other PKC isoforms remained unchanged. Using a multiplicity of infection of 100 plaque-forming units/myocyte, basal and cofactor-dependent PKC epsilon kinase activity was increased 28- and 90-fold, respectively, when compared to control. Myocyte basal fractional shortening and [Ca2+]i transient amplitude were both increased by 21% (P < 0.05 each) in Ad-PKC epsilon transfected myocytes when compared to Ad-LacZ transfected control myocytes. The positive inotropic effect of ET-1 in control myocytes was markedly blunted in PKC epsilon-overexpressing myocytes. CONCLUSION: Specific overexpression of PKC epsilon in rabbit ventricular myocytes increases basal myocyte contractility and [Ca2+]i transients, and modifies their responsiveness to ET-1.


Subject(s)
Cardiotonic Agents/pharmacology , Endothelin-1/pharmacology , Myocardial Contraction/physiology , Myocardium/enzymology , Protein Kinase C/metabolism , Adenoviridae/genetics , Animals , Calcium/metabolism , Cell Culture Techniques , Dose-Response Relationship, Drug , Gene Transfer Techniques , Genetic Vectors , Heart Ventricles/cytology , Heart Ventricles/enzymology , Myocardial Contraction/drug effects , Rabbits
6.
J Pharmacol Toxicol Methods ; 42(1): 21-30, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10715600

ABSTRACT

Investigations of the coupling between contraction and relaxation (contraction-relaxation [CRC] process) in isometric conditions are essential in determining whether pharmacologic interventions or cardiac diseases specifically modify isometric relaxation (intrinsic lusitropic effect) or change it in proportion with the accompanying changes in contractility (or inotropy). For this purpose, the CRC process is quantified by various indexes, derived from differentiation and/or curve fitting the whole or relaxation phase of the isometric twitch, one of the most used being tau, the time constant of the final iso(volu)metric phase of relaxation. Nevertheless, the possible redundancy and validity of such indexes have not been thoroughly investigated. Accordingly, we performed a pharmacologic evaluation of such indexes in isolated rabbit ventricular muscles isometrically contracting in vitro, using modifiers of either intracellular Ca(2)+ handling (nifedipine, ryanodine, 2,5-di-tert-butyl-benzohydroquinone, all negative inotropic compounds, and BAY K 8644, a positive inotropic drug), or myofibrillar Ca(2)+ sensitivity (CGP 48506, a Ca(2)+ sensitizer, and butanedione monoxime, a Ca(2)+ desensitizer, respectively positive and negative inotropic compounds). The isometric twitch in control conditions and in the presence of increasing concentration of each compound was analyzed to determine the classically used CRC and/or lusitropic indexes, derived either from single parameters such as the maximal rate or contraction and relaxation (+dT(max) and -dT(max), respectively), or from curve fitting of the whole, or part, of the twitch. As the rate of isometric relaxation is dependent on myofilament properties, we expected that compounds modifying myofibrillar Ca(2)+ sensitivity in an opposite direction (CGP 48506 vs butanedione monoxime) would be the only drugs exerting an intrinsic lusitropic and opposite effect on a validated CRC index. Results showed that (1) none of the tested compounds affected the slope of the linear relationship between peak twitch tension and dT(max), a previously assumed CRC index, sensitive only to myofibrillar Ca(2)+ sensitivity modifiers; (2) the lusitropic parameter B, derived from mathematical curve fitting of the whole isometric twitch, and the ratio +dT(max)/dT(max), exhibited similar drug- and dose-dependency, but no opposite sensitivity to CGP 48506 and BDM for either index; and (3) negative inotropic compounds dose-dependently slowed relaxation (and conversely for positive inotropes), whether the latter was quantified by the rate constant beta, derived from double exponential curve fitting of the whole relaxation phase, or by the time constants tau(L) and tau(E), derived from the curve fitting (logistic and monoexponential, respectively) of the final phase of relaxation. Nevertheless, the pharmacologicly induced changes in beta were statistically significant at lower concentrations and exhibited less individual variability, compared with the time constants. We demonstrate that intrinsic lusitropic changes can be quantified by the value of the slope of the relationship relating beta to peak isometric tension: the slope value was unchanged by Ca(2)+ handling modifiers, decreased by CGP 48506, and reversed by BDM (indicating number, negative, and positive intrinsic lusitropic effects respectively). Based on these data, we propose that the linear relationship between beta and peak isometric tension could be used a new method to assess whether pharmacologic interventions or cardiac diseases exert intrinsic effects on isometric relaxation.


Subject(s)
Myocardial Contraction/drug effects , 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester/pharmacology , Animals , Azocines/pharmacology , Calcium/metabolism , Dose-Response Relationship, Drug , In Vitro Techniques , Nifedipine/pharmacology , Rabbits
7.
Naunyn Schmiedebergs Arch Pharmacol ; 358(4): 474-82, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9826070

ABSTRACT

In the present study we have investigated the involvement of sensitized mice immunoglobulins and some electrophysiological alterations that participate to the antigenic sensitization-induced hyperreactivity of isolated mouse vas deferens. Active sensitization was performed by subcutaneous injection of egg albumen. Contractile responses to noradrenaline were isometrically recorded in the isolated vas deferens. Low external Na(+)-induced contractions and rapid cooling contractures were evaluated. Resting membrane potential (Er) and intracellular Na activity were measured in control and actively sensitized vas deferens by using conventional KCl-filled and Na(+)-sensitive microelectrodes respectively. Active sensitization-induced hyperreactivity to noradrenaline was reproduced by in vitro passive sensitization of control vas deferens with sensitized mice immunoglobulins. The inhibition of the nitric oxide synthesis by N-nitro-L-arginine methyl ester (L-NAME) did not change control vas deferens reactivity in vitro to noradrenaline and acetylcholine. Rapid cooling contractures, performed after lowering external Na+ concentration, were not altered by active sensitization. However, sensitization increased significantly the strength of the low external Na+-induced contractions. In control vas deferens Er was a mean of -49.2+/-0.3 mV (mean+/-SEM). Sensitization resulted in reduction of Er by 14 mV. In sensitized preparations, relative insensitivity of Er to ouabain, external K+ removal and cooling were observed. The intracellular Na+ activity was increased by about 40% in sensitized vas deferens. It is concluded that sensitization-induced hyperreactivity is mediated by immunoglobulins and produced smooth muscle cells depolarisation. The low Er of sensitized muscle may be partly the result of an increase in membrane permeability to Na+ which could interfere with intracellular Ca2+ homeostasis.


Subject(s)
Immunoglobulins/immunology , Muscle Contraction/drug effects , Sodium/metabolism , Vas Deferens/physiology , Animals , Calcium/metabolism , Homeostasis , Immunization , Male , Membrane Potentials , Mice , NG-Nitroarginine Methyl Ester/pharmacology , Norepinephrine/pharmacology , Sodium-Potassium-Exchanging ATPase/physiology , Vas Deferens/drug effects
8.
Muscle Nerve ; 21(11): 1530-2, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9771680

ABSTRACT

During postnatal development of extensor digitorum longus (EDL) muscle, sodium withdrawal contractures were observed during the first 6 days after birth, and not after this time. In regenerating EDL muscles, zero-Na contractures were demonstrated: (1) 7 days after bupivacaine injection, but not 14 or 90 days after this injection; (2) 7, 14, and 90 days after autotransplantation; and (3) 7, 14, and 90 days after the intervention in sliced muscles. The present findings emphasize the role of the denervation in the development of zero-Na contractures in the regenerating muscles and suggest that a calcium-sodium exchange across the sarcolemma may appear in these muscles.


Subject(s)
Contracture/physiopathology , Muscle, Skeletal/physiology , Muscle, Skeletal/transplantation , Regeneration/physiology , Sodium Chloride/pharmacology , Anesthetics, Local/pharmacology , Animals , Bupivacaine/pharmacology , Calcium/physiology , Contracture/chemically induced , Male , Muscle Contraction/drug effects , Muscle Contraction/physiology , Muscle Development , Muscle, Skeletal/growth & development , Rats , Rats, Wistar , Tetraethylammonium/pharmacology
9.
Can J Physiol Pharmacol ; 76(7-8): 802-6, 1998.
Article in English | MEDLINE | ID: mdl-10030462

ABSTRACT

Smooth muscles hyperresponsiveness is a common feature in anaphylaxis and allergic diseases. The aim of the present work was to investigate whether the enhanced reactivity of sensitized guinea-pig vas deferens was associated with changes in the resting membrane potential (Er) of the smooth muscle cells. Active sensitization was performed by subcutaneous injection of egg albumen. Er was measured in vitro in isolated vas deferens with conventional KCl-filled microelectrodes. Quantification of [3H]ouabain binding sites, measurements of 86Rb efflux, and measurements of Na and K contents were also performed. In normal physiological solution, at 35 degrees C, Er was a mean of -54.1+/-0.3 mV (mean +/- SEM) in control vas deferens. Sensitization resulted in depolarizing Er by about 7 mV. In control and sensitized preparations, the 3H-ouabain binding site concentration, the efflux of 86Rb, and the K content were similar. In guinea-pig vas deferens, active sensitization induced a partial depolarization of the resting membrane potential of the smooth muscle cells, which did not result from a downregulation of Na+ -K+ pump sites.


Subject(s)
Muscle, Smooth/immunology , Vas Deferens/immunology , Animals , Binding Sites , Guinea Pigs , In Vitro Techniques , Male , Membrane Potentials/immunology , Muscle, Smooth/metabolism , Muscle, Smooth/physiology , Ouabain/metabolism , Ovalbumin/immunology , Potassium/analysis , Rubidium/metabolism , Sodium/analysis , Sodium-Potassium-Exchanging ATPase/physiology , Vas Deferens/metabolism , Vas Deferens/physiology
10.
Can J Physiol Pharmacol ; 76(12): 1161-5, 1998 Dec.
Article in English | MEDLINE | ID: mdl-10326840

ABSTRACT

Sarcoplasmic reticulum (SR) calcium handling in diaphragm was compared between mdx mice (7-8 weeks old) and age-matched controls. The total SR Ca2+ load was released from the SR by rapidly cooling muscle bundles from 22 to -1 degree C. The plateau amplitude of the rapid cooling contracture (RCC) was considered as an index of the SR Ca2+ content. The steady-state RCC amplitude was significantly lower by 50% in mdx bundles mainly because of a decreased capacity of the dystrophic diaphragm to generate maximal tension. There was no significant difference between either RCC time to peak or the time to half-relaxation of the transient, spike-like, contractile response induced by muscle rewarming. The recovery process of RCC was studied by using a paired RCC protocol. In both groups, at the shortest interval (10 s) between two RCCs, the amplitude of the second RCC was decreased by 25% compared with the first RCC. Increasing the time interval led to progressive monoexponential recovery of the second RCC with similar time constants in control and mdx diaphragm. These results indicate that the dystrophic process does not significantly alter SR Ca2+ uptake nor Ca2+ redistribution within the muscular cell.


Subject(s)
Calcium-Transporting ATPases/physiology , Diaphragm/physiology , Muscle Contraction/physiology , Sarcoplasmic Reticulum/physiology , Animals , Mice , Mice, Inbred C57BL , Mice, Inbred mdx , Muscular Dystrophy, Animal
11.
Biochem Biophys Res Commun ; 240(3): 612-7, 1997 Nov 26.
Article in English | MEDLINE | ID: mdl-9398613

ABSTRACT

Following disruption of the nerve supply, extensor digitorum longus (EDL) and soleus (SOL) muscles in rats are known to exhibit alterations in excitation-contraction coupling. After total RNA isolation from the denervated and the contralateral control muscles performed at 25 and 50 days following denervation, RNase protection assays were carried out with four cDNA probes specific for the skeletal and cardiac isoforms of both the DHPR alpha 1-subunit and the RyR. Longterm denervation increased the expression of the mRNA for skeletal DHPR and skeletal RyR in SOL muscle, but it also significantly increased the expression of the mRNA for the cardiac isoform of the DHPR alpha 1 subunit in EDL muscle.


Subject(s)
Calcium Channels/genetics , Gene Expression Regulation/genetics , Muscle Denervation , Muscles/metabolism , Ryanodine Receptor Calcium Release Channel/genetics , Animals , Calcium Channels, L-Type , DNA Probes , Electrophoresis, Polyacrylamide Gel , Heart/innervation , Muscle, Skeletal/innervation , Muscle, Skeletal/metabolism , Muscles/innervation , Myocardium/metabolism , RNA, Antisense/genetics , RNA, Antisense/metabolism , Rats , Ribonucleases/metabolism
12.
Muscle Nerve ; 20(8): 1032-4, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9236796

ABSTRACT

Contractile responses were studied in autotransplanted and sliced extensor digitorum longus (EDL) muscles at different times after the surgical intervention (3 and 12 months). The contraction of autotransplanted and sliced EDL muscles remained strongly dependent upon the extracellular Ca2+ concentration ([Ca]o) 3 and even 12 months after the intervention. It could be suggested that in autotransplanted and sliced EDL muscles, neural control could be involved in the persistent dependence of contractility toward [Ca]o.


Subject(s)
Calcium/pharmacology , Muscle Contraction/drug effects , Muscle, Skeletal/physiology , Muscle, Skeletal/transplantation , Regeneration , Animals , Denervation , Dose-Response Relationship, Drug , Male , Muscle Contraction/physiology , Muscle Fibers, Skeletal/drug effects , Muscle Fibers, Skeletal/physiology , Muscle, Skeletal/innervation , Rats , Rats, Wistar , Transplantation, Autologous
13.
J Mol Cell Cardiol ; 29(7): 1903-13, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9236144

ABSTRACT

Using an isolated ferret heart preparation (Langendorff perfusion, perfusion pressure 90 mmHg), energy metabolism has been characterized in right and left ventricles from control and hypertrophied hearts. Hypertrophy was induced by pulmonary artery clipping for 30-45 days (right ventricle wall weight/body weight ratio increased by 70%). Myocardial contents of high energy phosphate compounds, glycogen and lactate, and the activities of some enzymes were biochemically measured in perfused hearts and also after ischemic arrest (30 min global ischemia). In hypertrophied right ventricles, PCr (-46%), Cr (-34%) levels, creatine kinase activity (-18%) were significantly decreased compared with control. ATP and Pi levels were not affected by hypertrophy. The adenylate energy charges were similar (0.85-0.86) in both types of heart. The activities of hexokinase (+26%), aldolase (+212%), pyruvate kinase (+14%) and glucose 6-phosphate dehydrogenase (+107%) were increased by hypertrophy. The LDH isozyme pattern was significantly changed such that LDH3 was decreased by 11%, and LDH4 and LDH5 were increased by a factor 1.4 and 2.9 respectively in hypertrophy. After 30 min of global ischemia, PCr level was decreased by 89 and 79% in control and hypertrophied ventricles respectively. ATP level was depressed by 41 in control and only by 21% in hypertrophied muscles. Altogether, the present data suggested that, in the adult ferret heart, the capacity for the ATP synthesis could be maintained during hypertrophy by the enhancement of the glycolytic pathway. The smaller decline of ATP after ischemia in hypertrophied tissue could be explained by a lower consumption of ATP in the hypertrophied compared to the control heart during the earliest period of ischemia.


Subject(s)
Cardiomegaly/metabolism , Myocardium/metabolism , Adenosine Triphosphate/metabolism , Age Factors , Animals , Creatine Kinase/metabolism , Energy Metabolism , Ferrets , Glycolysis , Phosphates/metabolism , Phosphocreatine/metabolism
14.
Pflugers Arch ; 433(3): 221-9, 1997 Jan.
Article in English | MEDLINE | ID: mdl-9064636

ABSTRACT

One of the the major properties of mature skeletal muscle is its ability to regenerate after injury. The purpose of the present study was to determine whether the expression of genes encoding the dihydropyridine receptor calcium channel (DHPR) and the ryanodine receptor (RyR), which play a critical role in excitation-contraction coupling, is regulated by skeletal muscle regeneration. The process of regeneration was induced by bupivacaine injection in surgically exposed rat extensor digitorum longus (EDL) muscle. After total RNA isolation from the injected and the contralateral control EDL muscles performed 3, 7, 15 and 30 days following injection, Northern blot and RNase protection assays were carried out with four cDNA probes specific for the skeletal and cardiac muscle isoforms of both the DHPR alpha1-subunit and the RyR. After 3 days, an initial precipitous decrease in the expression of the genes encoding the skeletal muscle isoforms of the DHPR and RyR was observed, followed by an increase. Moreover, regenerating skeletal muscle transiently expressed mRNA for the DHPR cardiac isoform, mainly at the beginning of regeneration. No expression of mRNA for the cardiac RyR was observed. Contraction experiments, performed using EDL muscle at the same times after bupivacaine injection, showed that twitch amplitude was markedly decreased in the absence of external calcium, but only during the early stages of regeneration. Similar findings in relation to expression of skeletal and cardiac muscle DHPR message were previously reported from experiments conducted during early developmental stages using fetal skeletal muscle and muscle cell cultures [Chaudhari N, Beam KG (1993) Dev Biol 155:507-515]. These results suggest that expression of the DHPR cardiac isoform in skeletal muscle could explain certain cardiac-like aspects of excitation-contraction coupling of regenerating skeletal muscle and developing skeletal muscle as well.


Subject(s)
Calcium Channels/metabolism , Muscle Proteins/metabolism , Muscle, Skeletal/metabolism , Regeneration/physiology , Animals , Calcium Channels, L-Type , Gene Expression/genetics , Male , Muscle, Skeletal/physiology , Rats , Ryanodine Receptor Calcium Release Channel
15.
Biochim Biophys Acta ; 1362(2-3): 201-7, 1997 Dec 31.
Article in English | MEDLINE | ID: mdl-9540851

ABSTRACT

The expression of isoform-specific dihydropyrine receptor-calcium channel (DHPR) alpha 1-subunit genes was investigated in mdx and control mouse diaphragm (DIA) and tibialis anterior (TA). RNase protection assays were carried out with a rat DHPR cDNA probe specific for skeletal muscle and a mouse DHPR cDNA probe specific for cardiac muscle. The level of expression of the gene encoding the cardiac DHPR was very weak in TA muscle from both control and mdx mice. Compared to TA, DIA expressed mRNA for the cardiac isoform at significantly higher levels, but mdx and control mouse DIA levels were similar to one another. In contrast, mRNA expression levels for the DHPR skeletal muscle isoform were lower in control DIA than TA. However, there was a dramatic increase in the expression for the DHPR skeletal muscle isoform in mdx DIA compared with control DIA, reaching the TA expression level, whereas dystrophy did not affect TA expression. [3H]-PN200-110 binding was used to further assess DIA DHPR expression at the protein level. The density of binding sites for the probe was not significantly affected in DIA muscles of mdx vs. control mice, but it was reduced in older mdx and control mice. The increase in DHPR mRNA levels without a consequent increase in DHPR protein expression could be secondary to possible enhanced protein degradation which occurs in mdx DIA. The altered DHPR expression levels found here do not appear to be responsible for the severe deficits in contractile function of the mdx DIA.


Subject(s)
Calcium Channels/genetics , Muscular Dystrophy, Animal/genetics , Animals , Calcium Channels/metabolism , Calcium Channels, L-Type , DNA, Complementary/genetics , Gene Expression , Isradipine/metabolism , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Muscle Proteins/genetics , Muscle Proteins/metabolism , Muscle, Skeletal/metabolism , Muscular Dystrophy, Animal/metabolism , Myocardium/metabolism , RNA, Messenger/genetics
16.
Muscle Nerve ; 19(11): 1421-8, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8874399

ABSTRACT

Two types of nerve lesions were performed at birth in rat extensor digitorum longus muscle: sciatic nerve transection (group A) and sciatic nerve crush allowing further reinnervation (group B). Contractile responses were then studied at different times after the denervation (7, 14, 30, and 60 days) and compared with control. Sixty days after the intervention, twitch and tetanic tensions remained dependent upon the extracellular Ca2+ concentration ([Ca]o) both in groups A and B. However, the depression of tensions following Ca2+ withdrawal was more important in group A. Sixty days after birth, in the presence of a Ca2+ channel blocker, Cd2+ (2 mmol L-1), a depression of the twitch tension was observed in group A (similarly to control 1-7 days postnatal muscles), whereas Cd2+ potentiated twitch tension in group B (similarly to control 14-60 days postnatal muscles). After glycerol treatment (detubulating procedure) performed in 60-day-old muscles, twitch tension was abolished in group B and control, whereas twitch tension was potentiated in group A. Thus, in developing muscles, neural control could be involved in the dependence of contractility toward [Ca]o. These results may be relevant for the understanding of the contractile properties of neuromuscular disorders with early onset.


Subject(s)
Birth Injuries/physiopathology , Calcium/pharmacology , Muscle Contraction/drug effects , Muscle, Skeletal/physiopathology , Sciatic Nerve/injuries , Toes , Animals , Birth Injuries/pathology , Cations, Divalent/pharmacology , Glycerol/pharmacology , Muscle, Skeletal/drug effects , Muscle, Skeletal/pathology , Rats , Rats, Wistar
17.
Acta Physiol Scand ; 158(3): 287-94, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8931772

ABSTRACT

The effects of perchlorate (6-40 mM) on contractile properties and depolarization-contraction coupling of diaphragm have been tested in muscle fibre bundles from 8 to 12-week-old mdx and control mice. Twitch contractile properties were affected by perchlorate to a similar extent in mdx and control muscles. In the absence of perchlorate, voltage-dependent parameters of both activation and inactivation were similar in mdx and control muscles. In both mdx and control muscles, perchlorate induced a shift of activation and inactivation curves toward more negative potentials. However, mdx diaphragm fibres were less sensitive to perchlorate in the low range of concentrations (6-12 mM). These results are unlikely to be due to a possible effect of the lack of dystrophin on the excitation-contraction coupling voltage sensor or ryanodine receptor. However, necrosis and regeneration occur in mdx mouse diaphragm and it is postulated that the presence of a high proportion of not fully mature fibres could contribute to the reduced sensitivity of mdx diaphragm fibres to perchlorate.


Subject(s)
Diaphragm/drug effects , Diaphragm/physiopathology , Muscle Contraction/drug effects , Muscular Dystrophy, Animal/physiopathology , Perchlorates/pharmacology , Sodium Compounds/pharmacology , Animals , Calcium Channels/drug effects , Calcium Channels/physiology , In Vitro Techniques , Male , Membrane Potentials/drug effects , Mice , Mice, Inbred C57BL , Mice, Inbred mdx , Muscle Proteins/drug effects , Muscle Proteins/physiology , Muscular Dystrophy, Animal/genetics , Potassium/pharmacology , Ryanodine Receptor Calcium Release Channel
18.
Muscle Nerve ; 19(8): 994-1002, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8756165

ABSTRACT

The effect of the intramuscular injection of bupivacaine hydrochloride on selected morphological characteristics and contractile properties of adult rat extensor digitorum longus muscle was studied. Recovery of normal fiber size was already present 30 days after bupivacaine injection and at 90 days after injection, values of the normalized twitch tension (mN/mg of tissue) and of the fatigue index approached those measured in control muscle, whereas the normalized tetanic tension remained 57% of control. At 7-30 days postinjection, twitch force was decreased by reducing [Ca2+]zero (substituted by Mg2+) or adding Co2+ (5 mmol/L-1). By contrast potentiation of the twitch was recorded in the presence of Cd2+ (2 mmol/L-1). Glycerol treatment only reduced, but did not eliminate twitches developed by muscles 7 days after injection. Present results emphasize the importance of the recovery process in the loss of the susceptibility of the contractile responses to extracellular calcium in bupivacaine-injected muscles. These data may be of interest in the evaluation of functional aspects of muscles in which injections of viral vector or autologous myoblasts have been performed.


Subject(s)
Bupivacaine/pharmacology , Calcium/pharmacology , Muscle Contraction/physiology , Muscles/drug effects , Regeneration/physiology , Animals , Caffeine/pharmacology , Glycerol/pharmacology , Male , Rats , Rats, Wistar
19.
Cardiovasc Res ; 31(5): 729-38, 1996 May.
Article in English | MEDLINE | ID: mdl-8763402

ABSTRACT

OBJECTIVE: To answer the questions: (a) What is the effect of hypertrophy on the intracellular pH (pHi) and buffering power of cardiac muscle, and (b) How does hypertrophy affect the ability of cardiac muscle to recover from intracellular acidosis induced by hypoxia. METHODS: In nominally HCO3(-)-free, HEPES-buffered Tyrode solution (35 degrees C), pHi and the intrinsic buffering power (beta i, measured in the presence of amiloride) was investigated using pH-sensitive microelectrodes. RESULTS: beta i was similar in both preparations (25 mM/pH unit at pHi 7.04). beta i was inversely related to pHi but the relationship was not significantly modified by hypertrophy. In the absence of amiloride, the time constant of pHi recovery (tau r) on removal of NH+4, was similar in normal (4.0 +/- 0.2 min, n = 5) and in hypertrophied muscles (4.3 +/- 0.3 min, n = 4; n.s.). In both preparations, net acid extrusion (JH) was similarly increased at lower values of pHi. Lowering temperature from 35 degrees to 22 degrees caused an alkalinization (0.15 pH units) of pHi. At 22 degrees C the mean values of pHi, beta i, tau r and JH were similar in normal and in hypertrophied muscles. At both temperatures and in both groups of preparations, recovery of pHi following hypoxia is approximately exponential. The time constant of recovery of pHi following hypoxia (tau rh) at 22 degrees C was not significantly different in hypertrophied muscles (7.2 +/- 0.9 min, n = 8) compared to controls (10.6 +/- 1.8 min, n = 13). However, at 35 degrees C, there was a significant difference in the mean values of tau rh which was smaller for hypertrophied muscles (3.9 +/- 0.3 min, n = 7) than for normal (7.1 +/- 1.1 min, n = 4, P < 0.005). For pHi 6.8-7.0, net acid extrusion in hypertrophied preparations was increased by a factor of 4 compared to normal. CONCLUSIONS: The intracellular buffering capacity and the pHi regulating capacity via Na+/H+ exchange are not significantly modified by right ventricular hypertrophy in ferret heart. The faster pHi recovery from hypoxia-induced acidification can be interpreted in terms of the role of lactate efflux in pHi control. The possible role of energy compartmentalization, its influence on the Na+ gradient and thus on pHi control after hypoxia, is discussed.


Subject(s)
Hydrogen/metabolism , Hypertrophy, Right Ventricular/metabolism , Intracellular Fluid/metabolism , Papillary Muscles/metabolism , Amiloride/pharmacology , Animals , Buffers , Ferrets , Hydrogen-Ion Concentration , Hypoxia/metabolism , In Vitro Techniques , Papillary Muscles/drug effects
20.
Acta Physiol Scand ; 156(4): 447-56, 1996 Apr.
Article in English | MEDLINE | ID: mdl-8732250

ABSTRACT

The effects of perchlorate (1-20 mM) on myofibrillar calcium responsiveness have been tested in Triton X-100-skinned fibre bundles from rat soleus (slow-twitch) and extensor digitorum longus (fast-twitch) skeletal muscles. In extensor digitorum longus and soleus, perchlorate dose-dependently shifted the pCa (-log[Ca2+])/tension relationship towards lower free calcium concentration (sensitizing effect) and maximal tension was unchanged. The degree of sensitization was greater in extensor digitorum longus than in soleus bundles. Reversibility after exposure to 12 mM perchlorate was complete in soleus but not in extensor digitorum longus muscles. In fact, the 'return' pCa/tension relationship in extensor digitorum longus was shifted to higher free calcium concentration (desensitizing effect) compared with control. Perchlorate (12 mM) also enhanced myofibrillar calcium responsiveness of frog semitendinosus skinned skeletal fibres. Assuming a passive distribution of perchlorate across the sarcolemma, this sensitizing effect is probably not involved in perchlorate-induced potentiation of contractile responses of intact muscles and thereby supports the specificity of perchlorate as an agonist of the excitation/calcium release sequence in skeletal muscle fibres.


Subject(s)
Calcium/physiology , Muscle Contraction/drug effects , Muscle, Skeletal/drug effects , Perchlorates/pharmacology , Animals , Dose-Response Relationship, Drug , Rats , Rats, Wistar
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