| 1. |
- Arheden, Håkan, et al.
(författare)
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Calcium sensitivity and energetics of contraction in skinned smooth muscle of the guinea pig taenia coli at altered pH
- 1989
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Ingår i: Pflügers Archiv. - 0031-6768. ; 413:5, s. 476-481
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Tidskriftsartikel (refereegranskat)abstract
- Calcium-sensitivity of contraction, force-velocity relation and ATP hydrolysis rate at different pH (6.2-7.8) were investigated in skinned smooth muscle preparations from the guinea pig taenia coli. Varied free-calcium levels were buffered by 4 mM BAPTA (1,2-bis(2-aminophenoxy)-ethane-N,N,N'N'-tetraacetic acid) which has calcium binding properties little affected by pH. A small increase of calcium-sensitivity of contraction was seen at pH 6.2 compared to 6.9 and 7.8 (ED50 shift of about 0.15 pCa units). The isometric force and Vmax in fibres activated either by calcium or by thiophosphorylation of the myosin light chains were each reduced by about 15% at pH 6.2 compared to 6.9 and 7.8. Following an isotonic quick release the shortening velocity decreases with time. This effect was more pronounced at pH 6.2 than at pH 6.9 or 7.8. The ATP hydrolysis rates in relaxed and thiophosphorylated fibres were essentially unaffected by alteration in pH between 6.2 and 7.8. Due to the lower force, energetic cost of force maintenance was thus somewhat increased at pH 6.2. These results suggest that pH alteration between 6.2 and 7.8 have effects on the properties of the contractile machinery of the smooth muscle in the skinned guinea pig taenia coli. The effects are however small and therefore probably of little functional importance over a pH range which should cover most cases of intracellular pH alteration under physiological or pathophysiological conditions.
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| 2. |
- Arheden, Håkan, et al.
(författare)
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Cross-bridge behaviour in skinned smooth muscle of the guinea-pig taenia coli at altered ionic strength
- 1988
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Ingår i: Journal of Physiology. - 1469-7793. ; 403, s. 539-558
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Tidskriftsartikel (refereegranskat)abstract
- 1. The effects of varied levels (25-300 mM) of ionic strength on mechanical properties and ATP hydrolysis rate of chemically skinned guinea-pig taenia coli fibres were investigated. 2. The tension development following activation by calcium (pCa 4 8), and relaxation following removal of calcium (pCa 9), were slower in 25 mm compared to 150 mm ionic strength. In fibres activated by thiophosphorylation of myosin light chains, by exposure to ATP-y-S, the tension development was rapid and independent of ionic strength. 3. The maximal shortening velocity (Vmax) was obtained from force-velocity relations determined by the quick-release method. The rate of ATP hydrolysis (JATP) was determined by measurement of pyruvate released from phosphoenolpyruvate (PEP). In order to obtain maximal Vmax and JATP at a Mg-ATP concentration of 1 mm, an ATP regenerating system was required. In thiophosphorylated fibres 2 mmphosphocreatine (PCr) or 3-2 mM-PEP were adequate for maximal Vmax and JATP respectively. In calcium-activated fibres 5 mM-PCr was required for maximal Vmax. 4. The isometric force of thiophosphorylated fibres showed a biphasic dependence on ionic strength with a maximum at 90 mm. Vmax was essentially unchanged between 50 and 200 mm ionic strength. At 25 mm ionic strength, isometric force and Vmax were decreased by, respectively, about 15 and 25%. At 250 mM ionic strength, isometric force and Vmax were decreased by, respectively, 47 and 33 %. 5. Vm.x decreased with decreasing [Mg-ATP]. At [Mg-ATP] less than 0 1 mm there was no difference in Vmax between 35 and 150 mM ionic strength. At 250 mM ionic strength Vmax was lower than that at 150 mm at all [Mg-ATP]. 6. JATP during contraction in thiophosphorylated fibres at 35, 150 and 250 mm ionic strength was respectively, 0-62, 0-98 and 0-93 ,umol g-1 min-'. The energetic tension cost (JATP/force) increased with ionic strength. 7. The force response to a quick stretch was investigated in the relaxed, contracted and rigor states at 25, 150 and 250 mm ionic strength. Stiffness in the relaxed state increased with speed of stretch and was higher the lower the ionic strength. In the contracted and rigor states, stiffness was also affected by ionic strength, but the relative effect in the contracted state was small. 8. The effects of ionic strength on the behaviour of the skinned smooth muscle fibre may involve an influence on the filament system, but are also compatible with an increased binding of smooth muscle myosin to actin at low ionic strength. The results from activated and rigor muscle suggest that ionic strength does not exclusively affect kinetics of rapid cross-bridge equilibria but may also influence mechanical properties of attached cross-bridge states.
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| 3. |
- Arheden, Håkan, et al.
(författare)
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Force-velocity relation and rate of ATP hydrolysis in osmotically compressed skinned smooth muscle of the guinea pig
- 1987
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Ingår i: Journal of Muscle Research and Cell Motility. - 0142-4319. ; 8:2, s. 151-160
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Tidskriftsartikel (refereegranskat)abstract
- Chemically skinned guinea pig taenia coli fibre bundles showed a concentration-dependent decrease in width when incubated in media containing Dextran T500 (0-0.2 g ml-1). The maximal reduction in width, observed at 0.2 g ml-1 dextran, was 32%. The effect was reversible upon removal of dextran. Isometric force was slightly increased (about 10%) at the lowest dextran concentration (0.025 g ml-1) but decreased at higher concentrations (40% decrease at 0.2 g/ml-1). The energetic tension cost (ATP turnover/force) was decreased by about 40% after dextran addition. Force development and relaxation were markedly slower in 0.1 g ml-1 and absent in 0.2 g ml-1 dextran. In isotonic quick-release experiments 0.025 g ml-1 dextran did not influence maximal shortening velocity (Vmax) and relative stiffness, whereas 0.1 g ml-1 markedly increased stiffness and decreased Vmax to about 27%. Vanadate induced relaxation in the activated muscle (pCa 4.5) both in the absence and presence (0.1 g ml-1) of dextran and increased the rate of relaxation (pCa 9) at 0.1 g ml-1 dextran. The isometric rate of crossbridge turnover, as reflected by the energetic tension cost and the rate of relaxation, was decreased at all degrees of osmotic compression. Crossbridge turnover rate during shortening (Vmax) was unaffected at an osmotic compression of 12% (width) but was decreased at higher compression (32%).
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| 4. |
- Arner, Anders, et al.
(författare)
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Effects of Ca2+ on force-velocity characteristics of normal and hypertrophic smooth muscle of the rat portal vein
- 1985
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Ingår i: Acta Physiologica Scandinavica. - 0001-6772. ; 124:4, s. 525-533
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Tidskriftsartikel (refereegranskat)abstract
- Portal hypertension was induced in rats by partial ligation of the hepatic branches of the portal vein. After 5 days of hypertension the portal veins were taken out and mounted for isometric and quick-release experiments. Portal veins from sham-operated normal rats served as controls. The ligated veins had an increased cross-sectional area, indicating smooth-muscle hypertrophy. Although the absolute magnitude of active force of these veins was increased, the active force per cross-sectional area was decreased, indicating an alteration in the properties of the contractile system. No difference in the Ca2+ concentration-response relations to K+-activated intact control and hypertrophic veins was found. In chemically skinned preparations, devoid of functional plasma membranes, the hypertrophic veins had similar Ca2+ sensitivity (in the presence of I microM calmodulin) but a lower force per cross-sectional area. Force-velocity relations were determined in K+-activated intact preparations. In control veins a reduction in extracellular Ca2+ was associated with a significant reduction in both isometric force and maximal shortening velocity (Vmax). In hypertrophic veins the decreased isometric force at maximal activation was associated with a low Vmax. A comparison between hypertrophic and submaximally stimulated control vessels showed corresponding Vmax and isometric force values. We conclude that the low isometric force of hypertrophic veins is associated with a lower rate of cross-bridge turnover. This could be an effect of alterations in the activation mechanisms or in the intrinsic properties of the contractile system itself.
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| 5. |
- Arner, Anders, et al.
(författare)
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Effects of calcium and substrate on force-velocity relation and energy turnover in skinned smooth muscle of the guinea-pig
- 1985
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Ingår i: Journal of Physiology. - 1469-7793. ; 360, s. 347-365
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Tidskriftsartikel (refereegranskat)abstract
- Mechanical properties and rate of ATP breakdown (JATP) have been determined in the chemically skinned guinea-pig taenia coli at 22 degrees C. The influence of varied [Ca2+], [Mg ATP] and muscle length were investigated. The shortening response after a step decrease in force (isotonic quick release) was highly curvilinear in the first 100-200 ms. This effect was shown to be a time-dependent response to the force step and not primarily caused by the shift along the length-force relation associated with shortening. Maximal shortening velocity (Vmax) decreased gradually following the release. At pCa (= -log [Ca2+]) 4.5, Vmax at 20 and 1000 ms after release was 0.49 +/- 0.07 and 0.041 +/- 0.004 (mean +/- S.E. of mean, n = 5) lengths s-1 respectively. Unloaded shortening velocity obtained from length steps of different magnitude (slack test) also showed a gradual decrease after the release, consistent with the isotonic release results. Increasing [Ca2+] from the relaxed state at pCa 9 (1 microM-calmodulin present) gave increased isometric force to a maximum at pCa 4.5. Half-maximal response was obtained at pCa 6.1. JATP at maximal force at pCa 4.5 was about 3 times the basal rate at pCa 9. The relation between JATP and force was highly non-linear, with a marked increase in JATP with little alteration in force at the highest [Ca2+]. When force was reduced to zero at pCa 4.5 by shortening the muscle to 0.3 L0 (L0 being the length giving maximal active force), JATP decreased by about 30%. At two levels of [Ca2+] giving similar force (pCa 5.75 and 4.5) the energetic tension cost obtained by length variations was lower at the low [Ca2+]. At pCa 6.0, Vmax and force were decreased to the same extent relative to their values at pCa 4.5. At pCa 5.75, where there was no reduction in force but a 25% decrease in isometric JATP, Vmax was unchanged relative to pCa 4.5. Force, Vmax and JATP were all dependent on [Mg ATP]. Half-maximal response was obtained at 0.1 mM for force and Vmax, and at 0.5 mM for JATP. The results are discussed in relation to a possible influence of both Ca2+ and Mg ATP on kinetic properties of the cross-bridge cycle.
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| 6. |
- Arner, Anders, et al.
(författare)
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Influence of ATP, ADP and AMPPNP on the energetics of contraction in skinned smooth muscle
- 1987
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Ingår i: Progress in Clinical and Biological Research. - 0361-7742. ; 245, s. 43-57
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Tidskriftsartikel (refereegranskat)abstract
- The contraction of smooth muscle is influenced by the substrate MgATP and the product MgADP. The effects on force, shortening velocity and ATP-turnover, are consistent with an influence on the kinetics of cross-bridge cycling. Part of these effects are mediated via an influence on the regulation of contraction by myosin light chain phosphorylation. Results from preparations activated by thiophosphorylation, show that MgATP and MgADP also interact directly at the cross-bridge level, and are consistent with MgADP acting as a competitive ATP-analogue. The slow shortening velocity and decreased rate of ATP-induced relaxation from rigor in the presence of MgADP, suggest an inhibition of cross-bridge detachment. The rate of ATP-turnover was decreased in the presence of the nonhydrolyzable ATP-analogue AMPPNP. These results may contribute to the characterization of the biochemical reactions in the structurally organized smooth muscle contractile system. In addition, the influence of MgATP and MgADP on smooth muscle contraction suggest that the concentrations of substrate and products, at the level of the contractile proteins, may constitute important regulatory factors in vivo under conditions, such as hypoxia and ischemia, associated with impaired cellular energy supply.
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| 7. |
- Hellstrand, Per, et al.
(författare)
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Myosin light chain phosphorylation and the cross-bridge cycle at low substrate concentration in chemically skinned guinea pig Taenia coli
- 1985
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Ingår i: Pflügers Archiv. - 0031-6768. ; 405:4, s. 323-328
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Tidskriftsartikel (refereegranskat)abstract
- Force-velocity relations, rate of ATP turnover (JATP), and phosphorylation of the 20,000 D myosin light chains (LC20) were measured in chemically skinned guinea pig Taenia coli. Relative LC20 phosphorylation at 3.2 mM MgATP was 17% in relaxed tissues at pCa 9, and increased with force at increasing [Ca2+] to a maximum of 67% at pCa 4.5. Force at pCa 4.5 was dependent on the MgATP concentration with a half-maximal response at about 0.1 mM. At 0.1 mM MgATP LC20 phosphorylation at pCa 4.5 was 38%. Both JATP and the maximal shortening velocity (Vmax) were reduced in 0.1 mM MgATP, to 32% and 43%, respectively, of their values at 3.2 mM MgATP. Low-MgATP thus inhibits both LC20 phosphorylation and the extent and rate of cross-bridge interaction. High levels of LC20 phosphorylation, independent of Ca2+ and MgATP concentrations, were obtained by treatment with ATP-gamma-S. Maximal force at 3.2 mM MgATP after LC20 thiophosphorylation was unchanged, whereas halfmaximal force occurred at 0.065 mM MgATP after thiophosphorylation, compared to 0.13 mM after activation by Ca2+. The contraction in thiophosphorylated preparations at low-MgATP (0.1 mM) was associated with submaximal Vmax (60%) and JATP (27%). The results show that LC20 phosphorylation is correlated with the degree of force development in the Ca2+ activated contraction, both when Ca2+ and MgATP concentrations are varied. The reduced force and rate of crossbridge turnover in low MgATP are however primarily mediated by an influence of MgATP on the cross-bridge cycle, which is separate from the effect on LC20 phosphorylation.
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| 8. |
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| 9. |
- Malmqvist, Ulf, et al.
(författare)
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Contractile properties during development of hypertrophy of the smooth muscle in the rat portal vein
- 1988
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Ingår i: Acta Physiologica Scandinavica. - 0001-6772. ; 133:1, s. 49-61
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Tidskriftsartikel (refereegranskat)abstract
- Structural and mechanical alterations during hypertrophy of the rat portal vein were investigated. Growth of the vessel was induced by a partial ligature of the vessel causing an increased transmural pressure. Vessel segments from animals kept with ligature for 1, 3, 5 and 7 days, were compared with vessels from sham-operated animals. Maximal active force and vessel cross-sectional area increased with time in the ligated group. On day 7, force and cross-sectional area at the optimal length, were markedly increased in the ligated group (21.1 +/- 1.0 mN, 0.55 +/- 0.04 mm2, n = 9) compared with the control vessels (11.7 +/- 1.0 mN, 0.30 +/- 0.02 mm2, n = 7). Light and electron microscopy of preparations fixed at optimal length showed that the amount of smooth muscle and the cross-sectional area of cell profiles were almost doubled in the ligated group on day 7, consistent with hypertrophy of the smooth muscle. The force per smooth muscle cell area was similar in the two groups (ligated: 132 +/- 15; control: 145 +/- 16 mN mm-2, n = 4-5). The maximal shortening velocity was significantly lower in the hypertrophied group (ligated: 0.28 +/- 0.02; control: 0.41 +/- 0.01 optimal length s-1, n = 6). In chemically skinned preparations, activated by maximal thiophosphorylation of the myosin light chains, force was higher in the ligated group compared to the controls but no difference in maximal shortening velocity was observed. In conclusion, the increased transmural pressure is associated with a rapid increase in the amount of smooth muscle in the portal vein. The mechanical data show that after 7 days the force generating ability of the contractile system has increased in proportion to the smooth muscle cell mass. The unaltered maximal shortening velocity in the skinned hypertrophied preparations suggests that the kinetic properties of the maximally activated contractile system are unaltered. The decreased maximal shortening velocity in the intact hypertrophied preparations may reflect alterations in the excitation-contraction coupling.
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| 10. |
- Sparrow, M P, et al.
(författare)
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Isoforms of myosin in smooth muscle
- 1987
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Ingår i: Progress in Clinical and Biological Research. - 0361-7742. ; 245, s. 67-79
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Tidskriftsartikel (refereegranskat)
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