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PubMed1995

Control of pulmonary vascular smooth muscle tone by sarcoplasmic reticulum Ca2+ pump blockers: thapsigargin and cyclopiazonic acid.

The effect of thapsigargin (TG) and cyclopiazonic acid (CPA) on the mechanical activity of the rat pulmonary artery were investigated. In chemically (beta-escin)-skinned arterial strips, application of TG (0.1-1 microM) or CPA (0.5-10 microM) prior and throughout the loading procedure of the internal Ca2+ stores (0.3 microM free Ca2+ ions for 8-10 min) concentration dependently inhibited the subsequent contractile response induced by noradrenaline (NA, 10 microM) or caffeine (25 mM). In intact strips repeatedly incubated in a Ca(2+)-containing solution (2.5 mM for 10 min), followed by incubation in a Ca(2+)-free solution 12 min before NA-stimulation, TG and CPA not only inhibited the NA-induced contraction but also increased the tension which appeared during the exposure time to Ca2+. The two phenomena developed with similar time courses. The increase in tension during the readmission of Ca2+ ions was not antagonized by verapamil (10 microM) or nifedipine (1 microM) but was blocked by La3+ (50 microM) and Co2+ (1 mM) ions. The amplitude of the verapamil-insensitive TG (or CPA)-induced contraction was dependent on the external [Ca2+] [0.1-10 mM, concentration for half maximal effect (EC50) = 0.85 mM], not modified by the reduction of the external [Na+] (from 130 to 10 mM) and decreased by depolarization of the strip using K(+)-rich (30-120 mM) solutions. Under the latter condition, 38 +/- 9 and 83 +/- 4% reduction (n = 5) was observed in the presence of 60 and 120 mM K+ respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

The variable coupling between force and myosin light chain phosphorylation in Triton-skinned chicken gizzard fibre bundles: role of myosin light chain phosphatase.

The mechanism responsible for the regulation of smooth muscle tone at low levels of myosin light chain (MLC) phosphorylation is still poorly understood. According to one model, so-called latchbridges, which contribute to force maintenance at low levels of MLC phosphorylation, are generated by dephosphorylation of attached and phosphorylated crossbridges. The model predicts that the force generated for a given level of MLC phosphorylation depends on the activity of the MLC phosphatase. We tested this hypothesis by reducing the activity of the phosphatase by at least 80% in two ways: inhibition with okadaic acid and extraction. Under both conditions, higher levels of MLC phosphorylation were required to support a given level of force, suggesting a decreased flux of attached phosphorylated to attached dephosphorylated crossbridges, as predicted by this model. Although, under both conditions, the relationship between force and MLC phosphorylation was shifted to the right, the curves did not superimpose as would have been expected if the phosphatase activity were the only determinant of the coupling between force and phosphorylation. In the extracted fibres, two more proteins, calponin and SM22, were significantly reduced in addition. Therefore, these proteins might be involved in modulating the relationship between force and MLC phosphorylation.

Animals↗
PubMed1993

Left ventricular pressure transmission to myocardial lymph vessels is different during systole and diastole.

In six open-thorax-anaesthetized dogs with paced hearts and a retrogradely cannulated epicardial lymph vessel, the sensitivity of myocardial lymph pressure to left ventricular pressure during systole and during diastole was determined. The lymph vessels were cannulated using PE-90 tubing, and lymph pressure was measured by connecting the cannula to a microtip pressure transducer. To obtain the systolic sensitivity, left ventricular pressure was changed by clamping the descending aorta, which caused left ventricular pressure to increase. The diastolic sensitivity was obtained from natural variation to left ventricular pressure caused by atrial contractions during induced long diastoles. The mean ratio of the pulse in lymph pressure to the pulse in left ventricular pressure was determined: systole: 0.069 +/- 0.013, n = 213, diastole: 0.76 +/- 0.16, n = 249 and, if possible, linear regression analysis between lymph and left ventricular pressure was performed. The systolic regression coefficients could be determined in six dogs and the diastolic coefficients in three dogs. During long diastoles lymph pressure variations are on average 76 per cent of those in the left ventricle. However, during systole, the sensitivity of lymph pressure to left ventricular pressure is more than ten times lower. It is not unlikely that the structural embedment of lymph vessels within the myocardium is such that volume variations by cardiac contraction are limited.

Animals↗
PubMed1996

A possible role for nitric oxide in the regulation of human ureteral smooth muscle tone in vitro.

There is ample evidence that nitric oxide (NO) is an important neurotransmitter in many tissues of the urogenital tract. The aim of the present study was to examine the possible role of NO in ureteral relaxation. Human ureteral rings were mounted in organ bath chambers and precontracted in KCl. Increasing doses of the NO donor linsidomine (SIN-1) were added with and without prior blockade of the NO/cGMP pathway by methylene blue and protein kinase (PK) inhibitors Rp-8-pCPT-cGMPS and RP-8-CPT-cAMPS. Electrical filed stimulation (EFS) was done before and after incubation with L-NOARG (NG-nitro-L-arginine) and TTX (tetratodoxin). For detection of neuronal NO synthase (NOS), ureters were stained immunohistochemically. Ureteral strips were dose dependently relaxed by SIN-1; preincubation with methylene blue and protein kinase G inhibitor significantly reduced the SIN-1-induced relaxations. No effects of L-NOARG and TTX on EFS-induced tone alterations were found. NOS-positive neuronal axons and nerve-ending-like structures were found in the muscular layers. Our in vitro findings suggest that ureteral relaxation may involve the NO pathway.

Cyclic GMP↗
PubMed1995

Insulin increases cyclic nucleotide content in human vascular smooth muscle cells: a mechanism potentially involved in insulin-induced modulation of vascular tone.

It has been suggested that insulin exerts a vasodilating effect, but the mechanisms involved are not completely understood. Since cyclic nucleotides mediate the vasodilation induced by endogenous substances, such as prostacyclin and nitric oxide, we aimed to investigate the influence of insulin (concentration range 240-960 pmol/l) on both cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) content in human vascular smooth muscle cells. Insulin dose-dependently increased both nucleotides (cAMP: from 0.7 +/- 0.1 to 2.6 +/- 0.4 pmol/10(6) cells, p = 0.0001; cGMP: from 1.3 +/- 0.2 to 3.4 +/- 0.7 pmol/10(6) cells, p = 0.033). This increase is receptor-mediated, since it was blunted when cells were preincubated with the tyrosine kinase inhibitor genistein. The effect of insulin remained significant (p = 0.0001) when preincubation with the phosphodiesterase inhibitor theophylline prevented cyclic nucleotide catabolism. The increase of cGMP was blunted when the cells were preincubated with the guanylate cyclase inhibitor methylene blue, and with the nitric oxide-synthase inhibitor NG-monomethyl-L-arginine. At all the concentrations tested, insulin potentiated the increase of cAMP induced by the stable prostacyclin analogue Iloprost (p = 0.0001), whereas only at 1920 pmol/l did it potentiate the cGMP increase induced by glyceryltrinitrate (p = 0.05). This study demonstrates that the vasodilating effects exerted by insulin may at least in part be attributable to an increase of both cGMP and cAMP via a receptor-mediated activation of adenylate and guanylate cyclases in human vascular smooth muscle cells and that the insulin effect on cGMP is mediated by nitric oxide.

Arterioles↗
PubMed1995

Ca2+ entry and vasoconstriction during osmotic swelling of vascular smooth muscle cells.

Exposure of aortic strips from guinea-pigs to hypotonic extracellular fluid is followed by marked vasoconstriction, which is inhibited by D-600 (3 microM), a blocker of voltage-sensitive Ca2+ channels. Conventional electrophysiology, patch-clamp studies, pH determination with 2',7' bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF) and Ca2+ measurements with Fura-2 have been performed on smooth muscle cells cultured either from rat or human aorta to further elucidate the underlying mechanisms. Exposure of the cells to a 25% hypotonic extracellular fluid leads to a rapid and fully reversible depolarization, paralleled by an increase of the selectivity and conductance of the cell membrane to Cl-, an acidification of the cytoplasm and an increase of intracellular Ca2+ concentration ([Ca2+]i). The latter is inhibited by the Ca2+ channel blocker D-600 (1-3 microM). It is concluded that osmotic cell swelling leads to the activation of an anion channel. The subsequent depolarization of the cell membrane activates voltage-sensitive Ca2+ channels which increases [Ca2+]i, thus stimulating the contraction of vascular smooth muscle cells.

Animals↗
PubMed1978

The microvibrations of the body, an index for examination stress.

Changes of whole-body microvibrations (MV) were quantitatively analyzed in 123 voluntary subjects during two examination situations and compared with the results obtained during the ensuing vacation. The force oscillations in the three space directions were measured during quiet bipedal standing and simultaneously recorded as a time function with the rectified and integrated force-time function (= rectified impulse). The rectified impulses related to body weight were used as a measure for the MV. Immediately before the examinations, all the subjects showed high values. These decreased significantly in the control measurement if the subjects passed the exams, but remained relatively high in the subjects who failed. In all three measurements, smokers and candidates who took tranquilizers before the exams show higher MV values than do the other subjects. Females are less tense than males, a difference particularly significant in the vertical (z) direction. The rectified impulse in this direction correlates positively with the estimated cardiac output. Body weight and body length show a positive correlation with the MV values in the horizontal forward-backward (x) and vertical (z) direction. The measurement of the whole-body MV enables quantitative evaluation of the complex load due to an examination situation. The hypothesis that different types of stress or personality affect given directions in space requires further investigation.

Adult↗
PubMed1980

Mean arterial pressure, O2-uptake, and muscle force time during dynamic and rhythmic-static exercise in men with high percentages of fast- and slow-twitch fibers.

The influence of different percentages of slow-twitch (ST) and fast-twitch (FT) fibers in vastus lateralis on the relationship between mean arterial pressure (MAP) and O2-uptake (VO2) with muscle force time (MIpm, 60 integral of 0 p X t) exerted on the pedal during upright cycling, or during rhythmic isometric contraction (RIC) (only one subjects) was studied in three high percent STUDIED IN THREE HIGH PERCENT ST men (average 78% ST) (ST group), and in three high percent FT men (average 75% FT) FT group). MAP, or MIpm, was higher and had a steeper linear regression in the FT group than in the ST group at or against the same absolute VO2 during cycling at 60 rpm, or 100 rpm. The relationship between MAP and MIpm, however, was almost the same between both in the FT and ST group, or between both at 60 and 100 rpm. During RIC the slope (delta y/delta x) of the linear regression line between MAP and MIpm was close to that during cycling, whereas the slope of VO2 vs. MIpm was much lower. In conclusion, the variations in fiber population in working muscle do not seem to have any effect on MAP. MAP increases with MIpm independent of fiber population and/or fitness (VO2 max).

Adult↗
PubMed1975

[Histological, histochemical, and ultrastructural findings in malignant hyperthermia (author's transl)].

Malignant hyperthermia is a rare but severe complication of modern anesthesia, induced by halothane and succinylcholine. The syndrome is characterized by a rapid sustained and extreme rise in body temperature associated with muscular rigidity, tachycardia, tachypnoea and cyanosis. The lethality is about 60%. The present paper describes the histological, histochemical and electron microscopical findings performed on muscle biopsies of 3 patients with malignant hyperthermia (1 patient died) and a so called risk patient. In all patients morphological findings consistent with a pre-existent myopathy were found. Histologically there were acute necrotic muscular fibers as well as in types I and II, variations in the fiber diameter and centralization of the nuclei. In two cases even fibers that had a normal aspect in HE slides, showed a pathologic pattern after phosphorylase reaction. In addition to acute rhabdomyolysis, electron-microscopic investigations revealed cystic expansion of the cisterns of the sarcoplasmic reticulum with a peculiar proliferation of the sarcolemma. In a degenerating mitochondrium, a crystalline inclusion was identified. These findings support the pathogenetic concept of Britt and coworkers of a functional defect in the calcium release or binding mechanism of sarcoplasmic reticulum. Since it is known that malignant hyperthermia has a familial predilection, it seems very important that clinical, biochemical, and morphological investigations be performed such as CPK estimations and muscular biopsies not only of the patients but also of the relatives in order to rule out this type of latent myopathy.

Adult↗
PubMed1985

The effects of LSD in the guinea-pig ileum. Inhibition of acetylcholine release and stimulation of smooth muscle.

The effects of lysergic acid diethylamide (LSD) on acetylcholine release and on smooth muscle tone were studied in the myenteric plexus-longitudinal muscle preparation of the guinea pig. LSD (0.01-10 microM) depressed in a concentration-dependent manner the electrically-evoked [3H]-acetylcholine outflow from strips preincubated with [3H]-choline. The maximal effect was a 45% inhibition by 1 microM LSD. The spontaneous outflow was not affected. Metitepine competitively antagonized (pA2 8.0) the LSD-induced reduction of the evoked outflow. Tolazoline and mepyramine did not affect the inhibitory action of LSD. The contractions in response to electrical stimulation were enhanced by 34% in the presence of 0.1 microM LSD. Other concentrations of LSD did not affect the twitches. LSD caused an increase in muscle tone which was antagonized non-competitively by mepyramine, metitepine and ketanserin. Ketanserin was a competitive antagonist against the histamine-induced contractions of the longitudinal muscle (pA2 8.49). The results suggest that LSD stimulates presynaptically located 5-HT receptors and thereby decreases the evoked acetylcholine release. In addition, LSD increases smooth muscle tone either directly through stimulation of H1 receptors or indirectly via histamine release.

Acetylcholine↗
PubMed1985

Effect of verapamil on the non-adrenergic response of the field stimulated rat vas deferens.

A comparison has been made, in the present study, between the effects of verapamil (reported to have smooth muscle depolarizing action) and K+ depolarization on the responses of noradrenaline, ATP and those of field stimulation on the vas deferens obtained from reserpinized rats. Field stimulation of the vas using single pulse (1 ms pulse width; supramaximal voltage) resulted in a fast twitch response reaching a maximum at 300 +/- 20 ms. Verapamil (6 X 10(-6) M) significantly potentiated this response. Verapamil potentiated the twitch component of the biphasic response resulting from field stimulation of the intrinsic nerves with repetitive pulses, while the tonic component was markedly inhibited. Verapamil enhanced the ATP (7 X 10(-5) M) response, while the phasic and tonic components of KCl (5.36 X 10(-2) M)-induced biphasic responses were nearly abolished. While the phasic component of the noradrenaline (7 X 10(-6) M) response remained unaltered in the presence of verapamil, the tonic component was markedly inhibited and rhythmicity following phasic component was markedly enhanced. Partial depolarization, achieved by increasing K+ concentration in the normal Krebs by two-fold i.e., to 11.8 mM, enhanced the responses of ATP, noradrenaline and the twitch resulted from the single pulse stimulation. The finding that verapamil potentiates the contractile response to exogenously applied ATP, which is believed to be the "noradrenergic" neurotransmitter in the vas deferens, suggests that this is the mechanism through which verapamil potentiates the twitch responses to field stimulation of the nerve supply.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1978

Series elastic properties of skinned muscle fibres in contraction and rigor.

Isometric tension of skinned fibres from the frog semitendinosus muscle is sigmoidally related to Ca2+ concentration between pCa 7 and 6. Stiffness measurements showed that the Ca2+-activated tension may be due to recruitment of attached cross-bridges. In the absence of ATP (rigor solution) the skinned fibre develops a rigor tension which reaches about 80-110% of the maximum Ca2+-activated tension. However, stiffness measurements showed that in rigor many more cross-bridges are attached to actin at any one moment than in contraction. It was concluded that the force per cross-bridge is 37% smaller in rigor than in contraction.

Actins↗
PubMed1976

Creep after loading in the relaxed and contracted smooth muscle (taenia coli of the guinea pig) under various osmotic conditions.

1. Experiments to investigate the creep phenomena of the taenia coli were performed in hypotonic (207 mosm) and hypertonic (447 or 463 mosm) media, in the relaxed (Ca2+-depletion + verapamil) as well as in the contracted (K2SO4-depolarized) state. They were compared with earlier results in isotonic media. The samples were standardized with respect to their volume (weight) before the beginning of the experiment. 2. Neither in the relaxed, nor in the contracted state was there a statistically significant difference in mean total length observed within each test series after loading with 9928 dyn for 2000 s (about 33 min) in the different osmotic media. We define total length as unloaded initial length lo + initial pure leastic extension lE + creep N. 3. In the contracted state the mean total lengths attained by elastic extension and creep are significantly greater than in the relaxed state in both hypotonic and isotonic media. The greater variance in the data for hypertonic solutions makes any statistical decision of this kind impossible. 4. The typical time course of creep (N) and particularly the hyperbolic dependance of N and dN/dt stated in an earlier paper could also be observed here under varied osmotic conditions. 5. The results suggest that the viscosity of the cell plasma is not significant for the time course and extent of creep in smooth muscles of vertebrates. Creep probably takes place in the solid structures of the cells: most particularly the contractile proteins may be involved.

Animals↗