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Dependence of cyclic-AMP induced relaxation on Ca2+ and calmodulin in skinned smooth muscle of guinea pigTaenia coli

  • Excitable Tissues and Central Nervous Physiology
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Abstract

cAMP (10−6–10−4 M) produced a dose-dependent relaxation of Ca2+-induced contraction in the guinea-pig taenia coli skinned with 1% Triton X-100. At 0.53 μM Ca2+ and 0.05 μM calmodulin (CaM), cAMP (10−4 M) produced a maximal relaxation of 75% (pH 6.7; 25°C). Increasing Ca2+ (0.8 μM) or CaM (0.37 μM) reduced cAMP-induced relaxation to 25 and 5% respectively. At high CaM (5 μM), cAMP-induced relaxation could be completely inhibited by as low as 0.25 μM Ca2+. Furthermore, small increases in Ca2+ or CaM could effectively reverse the cAMP-induced relaxation in the continuous presence of cAMP.

These results demonstrate that small modulations in the Ca2+-calmodulin activity have a strong effect on the ability of cAMP to produce a direct relaxing effect on the contractile proteins in skinned fiber. It is suggested that the effects of cAMP on the cellular mechanisms that lower cytoplasmic free Ca2+ concentration may act as the important determinants of the extent of the direct inhibitory effect of cAMP on the contractile elements. These two mechanisms may act in concert in this fashion to effect cAMP-induced relaxation in smooth muscle during β-adrenergic stimulation.

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References

  • Adelstein RS (1982) Calmodulin and the regulation of the actinmyosin interaction in smooth muscle and nonmuscle cells. Cell 30:349–350

    Google Scholar 

  • Adelstein RS, Hathaway DR (1979) Role of calcium and cyclic adenosine 3′:5″ monophosphate in regulating smooth muscle contraction. Am J Cardiol 44:783–787

    Google Scholar 

  • Adelstein RS, Conti MA, Hathaway DK, Klee CB (1978) Phosphorylation of smooth muscle myosin light chain kinase by the catalytic subunit of adenosine 3′:5′-monophosphate dependent protein kinase. J Biol Chem 253:8347–8350

    Google Scholar 

  • Arner A (1982) Mechanical characteristics of chemically skinned guinea-pig taenia coli. Pflügers Arch 395:277–284

    Google Scholar 

  • Breemen C van, Aaronson P, Loutzenheiser R, Meisheri K (1980) Ca2+ movements in smooth muscle. Chest 78 (Suppl):157–165

    Google Scholar 

  • Bülbring E, den Hertog A (1980) The action of isoprenaline on the smooth muscle of the guinea pig taenia coli. J Physiol 304:277–296

    Google Scholar 

  • Cassidy PS, Kerrick WG, Hoar PE, Malencik DA (1981) Exogenous calmodulin increases Ca2+ sensitivity of isometric tension activation and myosin phosphorylation in skinned smooth muscle. Pflügers Arch 392:115–120

    Google Scholar 

  • Casteels R, Raemaekers L (1979) The action of acetylcholine and catecholamines on an intracellular calcium store in the smooth muscle cells for the guinea-pig taenia coli. J Physiol 294:51–68

    Google Scholar 

  • Conti MA, Adelstein RS (1981) The relationship between calmodulin binding and phosphorylation of smooth muscle kinase by the catalytic subunit of 3′:5′-cAMP-dependent protein kinase. J Biol Chem 256:3178–3181

    Google Scholar 

  • Diamond J (1978) Role of cyclic nucleotides in control of smooth muscle contraction. In: George WJ, Ignarro LJ (eds) Adv Cycl Nucleo Res. Raven Press, New York, pp 327–340

    Google Scholar 

  • Endo M, Kitazawa T, Yagi S, Iino M, Kakuta Y (1977) Some properties of chemically skinned smooth muscle fibers. In: Casteels et al (eds) Excitation-contraction coupling in smooth muscle. Elsevier-North Holland, Amsterdam, pp 199–210

    Google Scholar 

  • Endo M, Kitazawa T, Yagi S (1980) Different features of the sarcoplasmic reticulum in cardiac and smooth muscles. In: Ebashis et al. (eds) Muscle contraction, its regulatory mechanism, Japan Sci Soc Press, Tokyo, pp 447–463

    Google Scholar 

  • Fabiato A (1982) Skinned fibers from skeletal, cardiac, and smooth muscles: Symposium. Federation Proc 41:2223–2224

    Google Scholar 

  • Gordon AR (1978) Contraction of detergent-treated smooth muscle. Proc Natl Acad Sci USA 75:3527–3530

    Google Scholar 

  • Guth K, Junge J (1982) Low Ca2+ impedes cross-bridge detachment in chemically skinned taenia coli. Nature 300:775–776

    Google Scholar 

  • Hartshorne DJ, Mrwa U (1982) Regulation of smooth muscle actomyosin. Blood Vessels 19:1–18

    Google Scholar 

  • Hirata M, Kuriyama H (1980) Does activation of cyclic AMP dependent phosphorylation induced by beta-adrenergic agent control the tone of vascular smooth muscle? J Physiol 307:143–161

    Google Scholar 

  • Iino M (1981) Tension responses of chemically skinned fiber bundles of the guinea-pig taenia coli under varied ionic environments. J Physiol 320:449–467

    Google Scholar 

  • Itoh T, Izumi H, Kuriyama H (1982) Mechanisms of relaxation induced by activation of β-adrenoceptors in smooth muscle cells of the guinea-pig mesenteric artery. J Physiol 326:475–493

    Google Scholar 

  • Janis RA, Barany K, Barany M, Sarimiento JG (1981) Association between myosin phosphorylation and contraction of rat uterine smooth muscle. Mol Pharmacol 1:3–11

    Google Scholar 

  • Kerrick WGL, Hoar PE (1981) Inhibition of smooth muscle tension by cyclic AMP-dependent protein kinase. Nature 292:253–255

    Google Scholar 

  • Martson SB (1982) The regulation of smooth muscle contractile proteins. Prog Biopphys Mol 41:1–41

    Google Scholar 

  • Meisheri KD, Breemen van C (1982) Effects of Beta-adrenergic stimulation on calcium movements in rabbit aortic smooth muscle: relationship with cyclic AMP. J Physiol 331:429–441

    Google Scholar 

  • Meisheri KD, McNeill JH (1979) Role of Ca in isoproterenol-induced increases in cAMP levels in rat uterus. Am J Physiol 6:C257–263

    Google Scholar 

  • Meisheri KD, Ruegg JC (1982) Relaxation of skinned taenia coli by micromolar concentration of cyclic AMP. J Muscle Res and Cell Motility 3:478

    Google Scholar 

  • Meisheri K, Pfitzer G, Ruegg JC (1983) Ca2+ and calmodulin dependence of cAMP-induced relaxation in chemically skinned smooth muscle. Pharmacologist 25 (3):141

    Google Scholar 

  • Miller JR, Stull JT (1982) Phosphorylation of myosin light chain kinase and relaxation of tracheal smooth muscle. Biophys J 37:53a (abstract)

    Google Scholar 

  • Miller JR, Silver PJ, Stull JT (1983) The role of myosin light chain kinase phosphorylation in beta-adrenergic relaxation of tracheal smooth muscle. Mol Pharmacol (in press)

  • Mrwa U, Troschka M, Ruegg JC (1979) Cyclic AMP-dependent inhibition of smooth muscle actomyosin. FEBS Lett 107:371–373

    Google Scholar 

  • Mueller E, Breemen van C (1979) Role of intracellular Ca2+ sequestration in beta-adrenergic relaxation of smooth muscle. Nature 281:682–683

    Google Scholar 

  • Nishikori K, Weisbrodt NW, Sherwood OD, Sanborn BM (1982) Relaxin alters rat uterine myosin light chain phosphorylation and related enzymatic activity. Endocrinology 111:1743–1745

    Google Scholar 

  • Nishikori K, Weisbrodt NW, Sherwood OD, Sanborn BM (1983) Effects of relaxin on rat uterine myosin light chain kinase activity and myosin light chain phosphorylation. J Biol Chem 258: 2468–2474

    Google Scholar 

  • Peterson JW (1982) Rate limiting steps in the tension development of freeze-glycerinated vascular smooth muscle. J Gen Physiol 79:437–452

    Google Scholar 

  • Portzehl H, Caldwell PC, Ruegg JC (1964) The dependence of contraction and relaxation of muscle fibers from the crab Maia Squinado on the internal concentration of free calcium ions. Biochim Biophys Acta 79:581–591

    Google Scholar 

  • Ruegg JC, Paul RJ (1982) Vascular smooth muscle: calmodulin and cyclic AMP dependent protein kinase alter calcium sensitivity in porcine carotid skinned fibers. Circ Res 50:394–399

    Google Scholar 

  • Ruegg JC, Sparrow MP, Mrwa U (1981) Cyclic AMP mediated relaxation of chemically skinned fibers of smooth muscle. Pflügers Arch 390:198–201

    Google Scholar 

  • Saida K (1982) Intracellular Ca release in skinned smooth muscle. J Gen Physiol 80:191–202

    Google Scholar 

  • Scheid CR, Honeyman TW, Fay FS (1979) Mechanism of betaadrenergic relaxation of smooth muscle. Nature 277:32–36

    Google Scholar 

  • Schneider M, Sparrow MP, Ruegg JC (1981) Inorganic phosphate promotes relaxation in chemically skinned smooth muscle of guinea-pig taenia coli. Experienta 37:980–982

    Google Scholar 

  • Silver PJ, Di Salvo J (1979) Adenosine 3:5-monophosphate mediated inhibition of myosin light chain phophorylation in bovine aortic actomyosin. J Biol Chem 254:9951–9954

    Google Scholar 

  • Silver PJ, Stull JT (1982) Regulation of myosin light chain and phosphorylase phosphorylation in tracheal smooth muscle. J Biol Chem 257:6145–6150

    Google Scholar 

  • Silver PJ, Holroyde MJ, Solaro RJ, DiSalvo J (1981) Ca, calmodulin, and cyclic AMP-dependent modulation of actin-myosin interactions in aorta. Biochim Biophys Acta 674:65–70

    Google Scholar 

  • Somlyo AP, Somlyo AV, Shuman H, Endo M (1982) Calcium and monovalent ions smooth muscle. Fed Proc 41:2883–2890

    Google Scholar 

  • Sparrow MP, Mrwa U, Hofmann F, Ruegg JC (1981) Calmodulin is essential for smooth muscle contraction. FEBS Lett 125:141–145

    Google Scholar 

  • Spedding M (1983) Direct inhibitory effects of some calcium antagonists and trifluoroperanzine on the contractile proteins in smooth muscle. Br J Pharmacol 79:225–231

    Google Scholar 

  • Teo TS, Wang TH, Wang JH (1973) Purification and properties of the protein activator of bovine heart cyclic adenosine 3:5-monophosphate phosphodiesterase. J Biol Chem 248:588–595

    Google Scholar 

  • Vincenzi FF (1981) Calmodulin pharmacology. Cell Calcium 2:387–409

    Google Scholar 

  • Walsh MP (1981) Calmodulin-dependent light chain kinases. Cell Calcium 2:333–352

    Google Scholar 

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Meisheri, K.D., Ruegg, J.C. Dependence of cyclic-AMP induced relaxation on Ca2+ and calmodulin in skinned smooth muscle of guinea pigTaenia coli . Pflugers Arch. 399, 315–320 (1983). https://doi.org/10.1007/BF00652759

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  • DOI: https://doi.org/10.1007/BF00652759

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