Pflügers Archiv

, Volume 416, Issue 6, pp 727–734

Intracellular free calcium concentration / force relationship in rabbit inferior vena cava activated by norepinephrine and high K+

  • Raouf A. Khalil
  • Cornelis van Breemen
Heart, Circulation, Respiration and Blood; Environmental and Exercise Physiology


The changes in isometric force and the underlying fluctuations in intracellular free calcium concentration ([Ca2+]i) were monitored simultaneously in thin sheets of rabbit inferior vena cava loaded with the fluorescent Ca2+ indicator fura-2. In resting tissues bathed in physiological saline solution, the estimated [Ca2+]i was approximately 105 nM. The α-adrenergic agonist norepinephrine (10 μM) caused an initial rise in [Ca2+]i to 264 nM during force development, which dropped to 216 nM during force maintenance. The maintained norepinephrine-induced increase in force and [Ca2+]i was reversed in Ca2+-free (2 mM EGTA) solution. Membrane depolarization by high K+ (80 mM) significantly increased [Ca2+]i to 234 nM. Compared to norepinephrine, high K+ caused about the same steady-state increase in [Ca2+]i, but a smaller increase in force. [Ca2+]i/force curves were constructed at different concentrations of extracellular Ca2+, with either norepinephrine or high K+ as a stimulant. The curve generated with norepinephrine was located to the left of that generated with high K+.

Key words

Vascular smooth muscle Calcium 


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  1. Ashida T, Blaustein MP (1987) Regulation of cell calcium and contractility in mammalian arterial smooth muscle: role of sodium-calcium exchange. J Physiol (Lond) 392:617–635Google Scholar
  2. Becker PL, Williams DA, Fay FS (1987) Cytosolic calcium transients in single muscle cells: kinetic characteristics. Biophys J 51:101 aGoogle Scholar
  3. Berridge MJ (1984) Inositol trisphosphate and diacylglycerol as second messengers. Biochem J 220:345–360Google Scholar
  4. Berridge MJ, Irvine RF (1984) Inositol triphosphate, a novel second messenger in cellular signal transduction. Nature 312:315–321Google Scholar
  5. Blinks JR, Wier WG, Hess P, Prendergast FG (1982) Measurement of Ca2+ concentrations in living cells. Prog Biophys Mol Biol 40:1–114Google Scholar
  6. Bolton TB (1979) Mechanisms of action of transmitters and other substances on smooth muscle. Physiol Rev 59:606–718Google Scholar
  7. Breemen C van, Aaronson P, Loutzenhiser R (1979) Na-Ca interactions in mammalian smooth muscle. Pharmacol Rev 30:167–208Google Scholar
  8. Breemen C van, Hwang K, Loutzenhiser R, Lukeman S, Yamamoto H (1985) Ca entry into vascular smooth muscle. In: Fleckenstein A, Breemen C van, Groz R, Hoffmeister F (eds) Cardiovascular effects of dihydropyridine-type calcium antagonists and agonists. Springer, Berlin Heidelberg New York, pp 58–71Google Scholar
  9. Carafoli E (1987) Intracellular calcium homeostasis. Annu Rev Biochem 56:395–433Google Scholar
  10. Caswell AH (1979) Methods of measuring intracellular calcium. Int Rev Cytol 56:145–181Google Scholar
  11. Chance B, Cohen P, Jobsis F, Schoener B (1962) Intracellular oxidation-reduction states in vivo. Science 137:499–508Google Scholar
  12. Chance B, Schoener B (1962) Correlation of oxidation-reduction changes of intracellular reduced pyridine nucleotides and changes in electroencephalogram of the rat in anoxia. Nature 195:956–958Google Scholar
  13. DeFeo TT, Morgan KG (1985) Calcium-force relationships as detected with aequorin in two different vascular smooth muscles of the ferret. J Physiol (Lond) 369:269–282Google Scholar
  14. DeFeo TT, Briggs GM, Morgan KG (1987) Ca2+ signals obtained with multiple indicators in mammalian vascular muscle cells. Am J Physiol 253:H1456-H1461Google Scholar
  15. Deth R, Breemen C van (1974) Relative contributions of Ca2+ influx and cellular Ca2+ release during drug induced activation of the rabbit aorta. Pflügers Arch 348:13–22Google Scholar
  16. Grynkiewicz P, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450Google Scholar
  17. Hai CM, Murphy RA (1989) Ca2+, crossbridge phosphorylation, and contraction. Annu Rev Physiol 51:285–298Google Scholar
  18. Hashimoto T, Hirata M, Itoh T, Kanmura Y, Kuriyama H (1986) Inositol-1,4,5-trisphosphate activates pharmaco-mechanical coupling in smooth muscle of the rabbit mesenteric artery. J Physiol (Lond) 370:605–618Google Scholar
  19. Kamm KE, Stull JT (1989) Regulation of smooth muscle contractile elements by second messengers. Annu Rev Physiol 51:299–313Google Scholar
  20. Khalil RA, Breemen C van (1988) Sustained contraction of vascular smooth muscle: Calcium influx or C-kinase activation? J Pharmacol Exp Ther 244:537–542Google Scholar
  21. Kitazawa T, Kobayashi S, Horiuti K, Somlyo AV, Somlyo AP (1989) Receptor-coupled, permeabilized smooth muscle. J Biol Chem 264:5339–5342Google Scholar
  22. Kobayashi S, Kanaide H, Nakamura M (1985) K+-depolarization induces a direct release of Ca2+ from intracellular storage sites in cultured vascular smooth muscle cells from rat aorta. Biochem Biophys Res Commun 129:877–884Google Scholar
  23. Morgan JP, Morgan KG (1984) Stimulus-specific patterns of intracellular calcium levels in smooth muscle of ferret protal vein. J Physiol (Lond) 351:155–167Google Scholar
  24. Nishimura J, Breemen C van (1989) Direct regulation of smooth muscle contractile elements by second messengers. Biochem Biophys Res Commun 163:929–935Google Scholar
  25. Nishimura J, Kolber, Breemen C van (1988) Norepinephrine and GTP-γ-s increase myofilament Ca2+ sensitivity in α-toxin permeabilized arterial smooth muscle. Biochem Biophys Res Commun 157:677–683Google Scholar
  26. Nishizuka Y (1984) The role of protein kinase C in cell surface signal transduction and tumor promotion. Nature 308:693–698Google Scholar
  27. Poenie M, Alderton J, Steinhardt R, Tsien RY (1986) Calcium rises abruptly and briefly throughout the cell on the onset of anaphase. Science 233:886–889Google Scholar
  28. Rembold CM, Murphy RA (1988) Myoplasmic [Ca2+] determines myosin phosphorylation in agonist-stimulated swine arterial smooth muscle. Circ Res 63:593–603Google Scholar
  29. Sato K, Ozaki H, Karaki H (1988) Changes in cytosolic calcium level in vascular smooth muscle strip measured simultaneously with contraction using fluorescent calcium indicator fura 2. J Pharmacol Exp Ther 246:294–300Google Scholar
  30. Smith JB, Dwyer SD, Smith L (1989) Decreasing extracellular Na+ concentration triggers inositol phosphate production and Ca2+ mobilization. J Biol Chem 264:831–837Google Scholar
  31. Somlyo AP, Himpens B (1989) Cell calcium and its regulation in smooth muscle. FASEB J 3:2266–2276Google Scholar
  32. Somlyo AP, Somlyo AV (1986) Electromechanical and pharmacomechanical coupling in vascular smooth muscle. J Pharmacol Exp Ther 159:129–145Google Scholar
  33. Somlyo AV, Bond M, Somlyo AP, Scarpa A (1985) Inositol trisphosphate-induced calcium release and contraction in vascular smooth muscle. Proc Natl Acad Sci USA 82:5231–5235Google Scholar
  34. Suematsu E, Hirata M, Hashimoto T, Kuriyama H (1984) Inositol 1,4,5-trisphosphate releases Ca2+ from intracellular store sites in skinned single cells of porcine coronary artery. Biochem Biophys Res Commun 120:481–485Google Scholar
  35. Ueno H, Sumimoto K, Hashimoto T, Hirata M, Kuriyama J (1987) Effect of procaine on pharmacomechanical coupling mechanisms activated by acetylcholine in smooth muscle cells of porcine coronary artery. Circ Res 60:356–366Google Scholar
  36. Williams DA, Fogarty KE, Tsien RY, Fay FS (1985) Calcium gradients in single smooth muscle cells revealed by the digital imaging microscope using fura-2. Nature 318:558–561Google Scholar
  37. Yamamoto H, Breemen C van (1985) Inositol 1,4,5-trisphosphate release calcium from skinned cultured smooth muscle cells. Biochem Biophys Res Commun 1:270–274Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Raouf A. Khalil
    • 1
  • Cornelis van Breemen
    • 1
  1. 1.Department of PharmacologyUniversity of Miami School of MedicineMiamiUSA

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