| 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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Activation of contraction and ATPase activity in intact and chemically skinned smooth muscle of rat portal vein. Dependence on Ca++ and muscle length
- 1983
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Ingår i: Circulation Research. - 0009-7330. ; 53:5, s. 695-702
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Tidskriftsartikel (refereegranskat)abstract
- The mechanical manifestations of muscle contraction (force development or shortening) are accompanied by an increased turnover of chemical energy (ATPase activity, JATP). In intact rat portal veins activated by high potassium medium to produce graded contractions at different levels of extracellular calcium, a linear dependence of oxygen consumption on force was found. The slope of the relation (metabolic tension cost) was higher during early stages than during late stages of contraction, possibly reflecting a transient high crossbridge ATP turnover rate. Chemically skinned (Triton X-100) rat portal vein preparations were used to study the energy turnover of the smooth muscle contractile system under constant activation. In these preparations, JATP increased on activation by calcium in the presence of 1 microM calmodulin, remained constant for maintained contractures, and decreased promptly on relaxation. Force declined with each repeated contraction at optimal calcium level (10(-4.5) M), but the relation between force and JATP remained invariant and agreed with that of the intact muscle during the transient of high ATP turnover. Calcium activation in the range 10(-9) to 10(-4.5) M caused a progressively steeper (nonlinear) increase in JATP with force. Length-force relations showed a lower relative force at muscle lengths below L0 in the intact than in the skinned tissue, possibly indicating depression of excitation-contraction coupling. The slope of the relation between JATP and force was lower when length was varied than when calcium was varied. At a length where no external force was produced, the activated muscle had a significantly higher JATP than while relaxed (10(-9) M Ca++). Together with previous results showing calcium dependence of the force-velocity relation, the present study indicates an influence of calcium on crossbridge kinetics in smooth muscle.
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| 5. |
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| 6. |
- 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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| 7. |
- Arner, Anders, et al.
(författare)
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Energy turnover and mechanical properties of resting and contracting aortas and portal veins from normotensive and spontaneously hypertensive rats
- 1981
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Ingår i: Circulation Research. - 0009-7330. ; 48:4, s. 539-548
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Tidskriftsartikel (refereegranskat)abstract
- Aortas and portal veins from spontaneously hypertensive rats (SHR) and matched normotensive Wistar-Kyoto rats (WKY) were studied with respect to their energy turnover and mechanical properties. Relaxed aortas fom SHR 16--17 weeks and 20--25 weeks of age were stiffer, had smaller circumference, and greater maximal active wall tension compared to WKY aortas. Active stress (forced/areas) was not different. Passive and active length-force relations of portal veins from 16 to 17-week-old SHR and WKY were not different. O2 consumption (JO2) and lactate production (JLA) were studied in aortas and portal veins from 20- to 25-week-old rats. In relaxed aortas, JO2 was 0.63 +/- 0.03 (n = 1) and 0.54 +/- 0.03 (n = 10) mu mol/min per g dry wt in SHR and WKY, respectively (P less than 0.05). On activation by high-K+ solution, JO2 increased with tension in a similar way in both groups. In contrast, JLA, about 0.85 mu mol/min per g, did not differ, and decreased with tension development. JO2 of relaxed portal veins, about 1.5 mu mol/min per g dry wt, and the increase in JO2 with contraction were not different between SHR and WKY, but the JO2-active stress relation was steeper in spontaneous activity than in Kv+ contractures. JLA was similar in SHR and WKY portal veins, about 1 mu mol/min per g, and unlike behavior of the aortas, it increased with tension. Thus it is evident that SHR show increased arterial metabolism, which is not accounted for by an increased energy demand of the contractile system.
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| 8. |
- 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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| 9. |
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| 10. |
- 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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