Inhibition of polyamine synthesis influences contractility of intestinal smooth muscle in culture

1997 ◽  
Vol 273 (1) ◽  
pp. C77-C84 ◽  
Author(s):  
K. Sward ◽  
B. O. Nilsson ◽  
P. Hellstrand
Keyword(s):  
Smooth Muscle ◽  
Contractile Activity ◽  
Tissue Concentrations ◽  
Polyamine Synthesis ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Adenosylmethionine Decarboxylase ◽  
High K ◽  
The Right ◽  
The Relationship

Smooth muscle strips from guinea pig ileum were cultured for 5 days and then tested for contractile properties to investigate whether endogenous polyamines influence excitation-contraction coupling. Inhibition of spermidine and spermine synthesis by culture in the presence of the adenosylmethionine decarboxylase (EC4.1.1.50) inhibitor CGP-48664 (1-10 microM) decreased spermidine and spermine levels by 50% and increased putrescine by 20-fold. After culture with 10 microM, but not 1 microM, CGP-48664, the relationship between extracellular Ca2+ concentration and force in high K(+)-depolarized strips was shifted to the right, and phasic contractile activity as well as sensitivity to muscarinic stimulation was enhanced. When spermidine and spermine (each 50 microM) were available for cellular uptake during culture in the presence of 10 microM CGP-48664, spermidine and spermine concentrations were increased, and the effect on Ca2+ sensitivity was reversed. In strips cultured with 0 or 1 microM CGP-48664 in the presence of 50 microM spermidine and 50 microM spermine, no effect on Ca2+ sensitivity was observed. Force development relative to intracellular Ca2+ concentration was decreased in CGP-48664 (10 microM)-treated strips. The results suggest that endogenous polyamines influence excitation-contraction coupling in smooth muscle, although overall tissue concentrations may not reflect the polyamine pools responsible for this effect.

Negative-feedback regulation of excitation-contraction coupling in gastric smooth muscle

1992 ◽  
Vol 263 (6) ◽  
pp. C1160-C1171 ◽  
Author(s):  
H. Ozaki ◽  
L. Zhang ◽  
I. L. Buxton ◽  
K. M. Sanders ◽  
N. G. Publicover
Keyword(s):  
Smooth Muscle ◽  
Negative Feedback ◽  
Contractile Activity ◽  
Slow Waves ◽  
Plateau Phase ◽  
Negative Feedback Regulation ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Pi Turnover ◽  
Resting Tension

The role of phosphatidylinositol (PI) turnover in excitation-contraction coupling was investigated in canine antral smooth muscle. Acetylcholine (ACh; 0.1-1 microM) transiently increased tissue levels of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and increased the amplitudes of the plateau phase of slow waves and associated Ca2+ transients and phasic contractions. ACh also increased basal concentrations of cytosolic Ca2+ ([Ca2+]c), but these changes were not associated with an increase in resting tension. ATP (0.3 mM) had similar effects on Ins(1,4,5)P3 levels, basal [Ca2+]c, and resting tension. However, in contrast to the effects of ACh, ATP transiently reduced the amplitude of the plateau phase of slow waves and reduced the amplitudes of associated Ca2+ transients and phasic contractions. We investigated the possibility that two products of PI turnover, diacylglycerol (DAG) and Ins(1,4,5)P3, might provide negative feedback to regulate Ca2+ entry during slow waves. 1) DAG is known to activate protein kinase C (PKC). Activation of PKC by phorbol 12,13-dibutyrate (PDBu, 0.5 microM) reduced the amplitude of the plateau phase of slow waves and corresponding Ca2+ transients and phasic contractions. Assay of PKC showed that ACh, ATP, and PDBu stimulated enzyme activity. 2) Ins(1,4,5)P3 is known to increase [Ca2+]c by release of Ca2+ from internal stores. Basal [Ca2+]c was also increased by elevated external K+, ionomycin, thapsigargin, or caffeine. Each of these compounds reduced the amplitude and duration of slow waves. Results suggest that products of PI turnover may provide negative-feedback control of Ca2+ influx during slow waves, tending to reduce the amplitude of phasic contractile activity in gastric muscles. Differences in responses to ACh and ATP can be explained by a G protein-dependent mechanism in which ACh suppresses the voltage dependence of Ca(2+)-activated K+ channels.

Long-term regulation of contractility and calcium current in smooth muscle

1997 ◽  
Vol 273 (5) ◽  
pp. C1714-C1720 ◽  
Author(s):  
Maria Gomez ◽  
Karl Swärd
Keyword(s):  
Smooth Muscle ◽  
Calf Serum ◽  
Cell Voltage ◽  
High K ◽  
Intracellular Ca ◽  
The Right ◽  
The Relationship ◽  
Cell Capacitance

Longitudinal smooth muscle strips from guinea pig ileum were cultured in vitro for 5 days, and the relationship between extracellular Ca2+ and force in high-K+ medium was evaluated. In strips cultured with 10% fetal calf serum (FCS), this relationship was shifted to the right (50% effective concentration changed by 2–3 mM) compared with strips cultured without FCS. The shift was prevented by inclusion of verapamil (1 μM) during culture and mimicked by ionomycin in the absence of FCS. The intracellular Ca2+ concentration ([Ca2+]i) during stimulation with high-K+solution or carbachol was reduced after culture with FCS, whereas the [Ca2+]i-force relationship was unaffected. Cells were isolated from cultured strips, and whole cell voltage-clamp experiments were performed. Maximum inward Ca2+ current (10 mM Ba2+), normalized to cell capacitance, was almost three times smaller in cells isolated from strips cultured with FCS. Culture with 1 μM verapamil prevented this reduction. These results suggest that increased [Ca2+]iduring culture downregulates Ca2+current density, with associated effects on contractility.

Calcium and excitation–contraction coupling in vascular smooth muscles

1982 ◽  
Vol 60 (4) ◽  
pp. 483-488 ◽  
Author(s):  
George B. Weiss
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Relative Size ◽  
Smooth Muscles ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
High K ◽  
Renal Vessels ◽  
Quantitative Separation ◽  
High Concentrations

The roles of Ca2+ in excitation–contraction coupling in vascular smooth muscle have been difficult to delineate, primarily because unambiguous association of specific Ca2+ components with morphologically defined cellular structures could not be attained. More recent use of washouts in La3+-substituted solutions at low temperature (to remove superficial Ca2+ and retain cellular Ca2+), Scatchard-coordinate plots (to identify incubation conditions appropriate for examining predominantly high or low affinity Ca2+ components), and high concentrations of Sr2+ (to remove high but not low affinity Ca2+) have facilitated qualitative and quantitative separation of different Ca2+ fractions. The release of high affinity Ca2+ elicited with norepinephrine and the increase in uptake of low affinity Ca2+ obtained with high K+ have been clearly demonstrated, and may directly measure or indirectly reflect changes in the level of intracellular free Ca2+. In other types of vascular smooth muscle (e.g., renal vessels, coronary arteries), similar Ca2+ components also appear to be present, but their relative size and functional importance for regulation of contractile responsiveness can differ.

scholarly journals Impaired left ventricular mechanical and energetic function in mice after cardiomyocyte-specific excision of Serca2

2014 ◽  
Vol 306 (7) ◽  
pp. H1018-H1024 ◽  
Author(s):  
N. T. Boardman ◽  
J. M. Aronsen ◽  
W. E. Louch ◽  
I. Sjaastad ◽  
F. Willoch ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Left Ventricular ◽  
Lv Function ◽  
Mechanical Function ◽  
Transport Mechanisms ◽  
Working Capacity ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Plasmic Reticulum ◽  
The Relationship

Sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA)2 transports Ca2+ from the cytosol into the sarcoplasmic reticulum of cardiomyocytes and is essential for maintaining myocardial Ca2+ handling and thus the mechanical function of the heart. SERCA2 is a major ATP consumer in excitation-contraction coupling but is regarded to contribute to energetically efficient Ca2+ handling in the cardiomyocyte. Previous studies using cardiomyocyte-specific SERCA2 knockout (KO) mice have demonstrated that decreased SERCA2 activity reduces the Ca2+ transient amplitude and induces compensatory Ca2+ transport mechanisms that may lead to more inefficient Ca2+ transport. In this study, we examined the relationship between left ventricular (LV) function and myocardial O2 consumption (MV̇o2) in ex vivo hearts from SERCA2 KO mice to directly measure how SERCA2 elimination influences mechanical and energetic features of the heart. Ex vivo hearts from SERCA2 KO hearts developed mechanical dysfunction at 4 wk and demonstrated virtually no working capacity at 7 wk. In accordance with the reported reduction in Ca2+ transient amplitude in cardiomyocytes from SERCA2 KO mice, work-independent MV̇o2 was decreased due to a reduced energy cost of excitation-contraction coupling. As these hearts also showed a marked impairment in the efficiency of chemomechanical energy transduction (contractile efficiency, i.e, work-dependent MV̇o2), hearts from SERCA2 KO mice were found to be mechanically inefficient. This ex vivo evaluation of mechanical and energetic function in hearts from SERCA2 KO mice brings together findings from previous experimental and mathematical modeling-based studies and demonstrates that reduced SERCA2 activity not only leads to mechanical dysfunction but also to energetic dysfunction.

Increased O2 cost of basal metabolism and excitation-contraction coupling in hearts from type 2 diabetic mice

2009 ◽  
Vol 296 (5) ◽  
pp. H1373-H1379 ◽  
Author(s):  
Neoma Boardman ◽  
Anne D. Hafstad ◽  
Terje S. Larsen ◽  
David L. Severson ◽  
Ellen Aasum
Keyword(s):  
Cardiac Metabolism ◽  
Therapeutic Benefit ◽  
Basal Metabolism ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Direct Measurements ◽  
The Difference ◽  
The Relationship ◽  
Type 2 Diabetic

We have reported previously that hearts from type 2 diabetic ( db/ db) mice show decreased cardiac efficiency due to increased work-independent myocardial O2 consumption (unloaded MV̇o2), indicating higher O2 use for nonmechanical processes such as basal metabolism (MV̇o2BM) and excitation-contraction coupling (MV̇o2ECC). Although alterations in cardiac metabolism and/or Ca2+ handling may contribute to increased energy expenditure in diabetic hearts, direct measurements of the O2 cost for these individual processes have not been determined. In this study, we 1) validate a procedure for measuring unloaded MV̇o2 directly (MV̇o2unloaded) and for determining MV̇o2BM and MV̇o2ECC separately in isolated perfused mouse hearts and 2) determine O2 cost for these processes in hearts from db/ db mice. Unloaded MV̇o2, extrapolated from the relationship between cardiac work (measured as pressure-volume area, PVA) and MV̇o2, was found to correspond with MV̇o2 measured directly in unloaded retrograde perfused hearts (MV̇o2unloaded). MV̇o2 in K+-arrested hearts was defined as MV̇o2BM; the difference between MV̇o2unloaded and MV̇o2BM represented MV̇o2ECC. This procedure was validated by demonstrating that elevations in perfusate fatty acid (FA) and/or Ca2+ concentrations resulted in changes in either MV̇o2BM and/or MV̇o2ECC. The higher MV̇o2unloaded in db/ db mice was due to both a higher MV̇o2BM and MV̇o2ECC. Elevation of glucose and insulin decreased FA oxidation and reduced both MV̇o2unloaded and MV̇o2BM. In conclusion, this study provides direct evidence that MV̇o2BM and MV̇o2ECC are elevated in diabetes and that acute metabolic interventions can have a therapeutic benefit in diabetic hearts due to a MV̇o2-lowering effect.

Contractile and Ca2+-handling properties of the right ventricular papillary muscle in the late-gestation sheep fetus

2006 ◽  
Vol 101 (3) ◽  
pp. 728-733 ◽  
Author(s):  
T. N. Spencer ◽  
K. J. Botting ◽  
J. L. Morrison ◽  
G. S. Posterino
Keyword(s):  
Right Ventricular ◽  
Papillary Muscle ◽  
Late Gestation ◽  
Postnatal Life ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Force Development ◽  
Uptake Rates ◽  
Significant Difference ◽  
The Right

The force-generating capacity of cardiomyocytes rapidly changes during gestation and early postnatal life coinciding with a transition in cardiomyocyte nucleation in both mice and rats. Changes in nucleation, in turn, appear to coincide with important changes in the excitation-contraction coupling architecture. However, it is not clear whether similar changes are observed in other mammals in which this transition occurs prenatally, such as sheep. Using small (70–300 μM diameter) chemically skinned cardiomyocyte bundles from the right ventricular papillary muscle of sheep fetuses at 126–132 and 137–140 days (d) gestational age (GA), we aimed to examine whether changes in cardiomyocyte nucleation during late gestation coincided with developmental changes in excitation-contraction coupling parameters (e.g., Ca2+ uptake, Ca2+ release, and force development). All experiments were conducted at room temperature (23 ± 1°C). We found that the proportion of mononucleate cardiomyocytes decreased significantly with GA (126–132d, 45.7 ± 4.7%, n = 7; 137–140d, 32.8 ± 1.6%, n = 6; P < 0.05). When we then examined force development between the two groups, there was no significant difference in either the maximal Ca2+-activated force (6.73 ± 1.54 mN/mm2, n = 14 vs. 6.55 ± 1.25 mN/mm2, n = 7, respectively) or the Ca2+ sensitivity of the contractile apparatus (pCa at 50% maximum Ca2+-activated force: 126–132d, 6.17 ± 0.06, n = 14; 137–140d, 6.24 ± 0.08, n = 7). However, sarcoplasmic reticulum (SR) Ca2+ uptake rates (but not Ca2+ release) increased with GA ( P < 0.05). These data reveal that during late gestation in sheep when there is a major transition in cardiomyocyte nucleation, SR Ca2+ uptake rates increase, which would influence total SR Ca2+ content and force production.

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