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Regulation of membrane excitability by intracellular pH (pHi) changers through Ca2+-activated K+ current (BK channel) in single smooth muscle cells from rabbit basilar artery

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Abstract

Employing microfluorometric system and patch clamp technique in rabbit basilar arterial myocytes, regulation mechanisms of vascular excitability were investigated by applying intracellular pH (pHi) changers such as sodium acetate (SA) and NH4Cl. Applications of caffeine produced transient phasic contractions in a reversible manner. These caffeine-induced contractions were significantly enhanced by SA and suppressed by NH4Cl. Intracellular Ca2+ concentration ([Ca2+]i) was monitored in a single isolated myocyte and based the ratio of fluorescence using Fura-2 AM (R 340/380). SA (20 mM) increased and NH4Cl (20 mM) decreased R 340/380 by 0.2 ± 0.03 and 0.1 ± 0.02, respectively, in a reversible manner. Caffeine (10 mM) transiently increased R 340/380 by 0.9 ± 0.07, and the ratio increment was significantly enhanced by SA and suppressed by NH4Cl, implying that SA and NH4Cl may affect [Ca2+]i (p < 0.05). Accordingly, we studied the effects of SA and NH4Cl on Ca2+-activated K+ current (IKCa) under patch clamp technique. Caffeine produced transient outward current at holding potential (V h) of 0 mV, caffeine induced transient outward K+ current, and the spontaneous transient outward currents were significantly enhanced by SA and suppressed by NH4Cl. In addition, IKCa was significantly increased by acidotic condition when pHi was lowered by altering the NH4Cl gradient across the cell membrane. Finally, the effects of SA and NH4Cl on the membrane excitability and basal tension were studied: Under current clamp mode, resting membrane potential (RMP) was −28 ± 2.3 mV in a single cell level and was depolarized by 13 ± 2.4 mV with 2 mM tetraethylammonium (TEA). SA hyperpolarized and NH4Cl depolarized RMP by 10 ± 1.9 and 16 ± 4.7 mV, respectively. SA-induced hyperpolarization and relaxation of basal tension was significantly inhibited by TEA. These results suggest that SA and NH4Cl might regulate vascular tone by altering membrane excitability through modulation of [Ca2+]i and Ca2+-activated K channels in rabbit basilar artery.

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References

  1. Aickin CC (1984) Direct measurement of intracellular pH and buffering power in smooth muscle cells of guinea pig vas deference. J Physiol 349:571–585

    PubMed  CAS  Google Scholar 

  2. Austin C, Wray S (1993) Extracellular pH signals affect rat vascular tone by rapid transduction into intracellular pH changes. J Physiol 466:1–8

    PubMed  CAS  Google Scholar 

  3. Austin C, Wray S (2000) Interations between Ca2+ and H+ and functional consequences in vascular smooth muscle. Circ Res 86:355–363

    PubMed  CAS  Google Scholar 

  4. Bae YM, Kim KS, Park JK, Ko E, Ryu SY, Baek HJ, Lee SH, Ho WK, Earm YE (2001) \( {\text{Ca}}^{{{\text{2 + }}}}_{{\text{i}}} \)-dependent membrane currents in vascular smooth muscle cells of the rabbit. Life Sci 69:2451–2466

    Article  PubMed  CAS  Google Scholar 

  5. Batlle DC, Peces R, LaPointe MS, Ye M, Daugirdas JT (1993) Cytosolic free calcium regulation in response to acute changes in intracellular pH in vascular smooth muscle. Am J Physiol 264(4 Pt 1):C932–C943

    PubMed  CAS  Google Scholar 

  6. Benham CD, Bolton TB, Lang RJ, Takewaki T (1985) The mechanism of action of Ba2+ and TEA on single Ca2+-activated K+-channels in arterial and intestinal smooth muscle cell membranes. Pflügers Arch 403:120–127

    Article  PubMed  CAS  Google Scholar 

  7. Berger MG, Vandier C, Bonnet P, Jackson WF, Rusch NJ (1998) Intracellular acidosis differentially regulates KV channel in coronary and pulmonary vascular muscle. Am J Physiol 275:H1351–H1359

    PubMed  CAS  Google Scholar 

  8. Brayden JE, Nelson MT (1992) Regulation of arterial tone by activation of calcium-dependent potassium channels. Science 256:532–535

    Article  PubMed  CAS  Google Scholar 

  9. Carl A, Lee HK, Sanders KM (1996) Regulation of ion channels in smooth muscles by calcium. Am J Physiol 271:C9–C34

    PubMed  CAS  Google Scholar 

  10. Carr P, McKinnon W, Poston L (1995) Mechanisms of pHi control and relationships between tension and pHi in human subcutaneous small arteries. Am J Physiol 268:C580–C589

    PubMed  CAS  Google Scholar 

  11. Danthuluri NR, Deth RC (1989) Effects of intracellular alkalinization on resting and agonist-induced vascular tone. Am J Physiol 256:H867–H875

    PubMed  CAS  Google Scholar 

  12. Danthuluri NR, Kim D, Brock TA (1990) Intracellular alkalinization leads to Ca2+ mobilization from agonist-sensitive pools in bovine aortic endothelial cells. J Biol Chem 265:19071–19076

    PubMed  CAS  Google Scholar 

  13. Elmoselhi AB, Blennerhassett M, Samson SE, Grover AK (1995) Properties of the sarcoplasmic reticulum Ca2+ pump in coronary artery skinned smooth muscle. Mol Cell Biochem 15:149–155

    Article  Google Scholar 

  14. Faraci FM, Breese KR, Heistad DD (1994) Cerebral vasodilation during hypercapnia. Role of glibenclamide-sensitive potassium channels and nitric oxide. Stroke 25:1679–1683

    PubMed  CAS  Google Scholar 

  15. Gaskell WH (1880) On the tonicity of the heart and blood vessels. J Physiol 3:48–75

    PubMed  CAS  Google Scholar 

  16. Harder DR (1984) Pressure-dependent membrane depolarization in cat middle cerebral artery. Circ Res 55:197–202

    PubMed  CAS  Google Scholar 

  17. Harpern MJ, Mulvany M (1997) Tension responses to small length of vascular smooth muscle cells. J Physiol 265:21–23

    Google Scholar 

  18. Hathaway DR, March KL, Lash JA, Adam LP, Wilensky RL (1991) Vascular smooth muscle. A review of the molecular basis of contractility. Circulation 83(2):382–290

    Google Scholar 

  19. Hayabuchi Y, Nakaya Y, Matsuoka S, Kuroda Y (1998) Effect of acidosis on Ca2+-activated K+ channels in cultured porcine coronary artery smooth muscle cells. Pflügers Arch 436:509–514

    Article  PubMed  CAS  Google Scholar 

  20. Horn R, Marty A (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol 92:145–159

    Article  PubMed  CAS  Google Scholar 

  21. Iino M (1990) Calcium release mechanisms in smooth muscle. Jpn J Pharmacol 54:345–354

    PubMed  CAS  Google Scholar 

  22. Iino S, Hayashi H, Saito H, Tokuno H, Tomita T (1994) Effects of intracellular pH on calcium currents and intracellular calcium ions in the smooth muscle of rabbit portal vein. Exp Physiol 79:669–680

    PubMed  CAS  Google Scholar 

  23. Jensen PE, Hughes A, Boonen HC, Aalkjær C (1993) Membrane potential, and [Ca2+]i during activation of rat mesenteric small arteries with norepinephrine, potassium, aluminum fluoride, and phorbol ester. Effects of changes in pHi. Circ Res 73:314–324

    PubMed  CAS  Google Scholar 

  24. Kang TM, So I, Kim KW (1995) Caffeine- and histamine-induced oscillations of K(Ca) current in single smooth muscle cells of rabbit cerebral artery. Pflügers Arch 431:91–100

    Article  PubMed  CAS  Google Scholar 

  25. Kang TM, So I, Uhm DY, Kim KW (1997) Two types of voltage-dependent outward potassium current in smooth muscle cells of rabbit basilar artery. Korean J Physiol Pharmacol 1:169–183

    CAS  Google Scholar 

  26. Kim SJ, Kim JK, So I, Suh SH, Lee SJ, Kim KW (1998) Characterization of Ca2+ stores in rabbit cerebral artery myocytes. Korean J Physiol Pharmacol 2:313–322

    CAS  Google Scholar 

  27. Klockner U, Isenberg G (1994) Intracellular pH modulates the availability of vascular l-type Ca2+ channels. J Gen Physiol 103:647–663

    Article  PubMed  CAS  Google Scholar 

  28. Kume H, Takagi K, Satake T, Tokuno H, Tomita T (1990) Effects of intracellular pH on calcium-activated potassium channels in rabbit tracheal smooth muscle. J Physiol 424:445–457

    PubMed  CAS  Google Scholar 

  29. Lindauer U, Vogt J, Schuh-Hofer S, Dreier JP, Dirnagl U (2003) Cerebrovascular vasodilation to extraluminal acidosis occurs via combined activation of ATP-sensitive and Ca2+-activated potassium channels. J Cereb Blood Flow Metab 23(10):1227–1238

    Article  PubMed  CAS  Google Scholar 

  30. Nagasaki K, Kasai M (1984) Channel selectivity and gating specificity of calcium-induced calcium release channel in isolated sarcoplasmic reticulum. J Biochem 96:1769–1775

    PubMed  CAS  Google Scholar 

  31. Nagesetty R, Paul RJ (1994) Effects of pHi on isometric force and [Ca2+]i in porcine coronary artery smooth muscle. Circ Res 75:990–998

    PubMed  CAS  Google Scholar 

  32. Nelson MT, Patlak JB, Worley JF, Standen NB (1990) Calcium channels, potassium channels and voltage-dependence of arterial smooth muscle tone. Am J Physiol 259:C3–C18

    PubMed  CAS  Google Scholar 

  33. Nelson MT (1993) Ca2+-activated potassium channels and ATP-sensitive potassium channels as modulators of vascular tone. Trends Cardiovasc Med 3:54–60

    Article  CAS  Google Scholar 

  34. Nelson MT, Cheng H, Rubart M, Santana LF, Bonev AD, Knot HJ, Lederer WJ (1995) Relaxation of arterial smooth muscle by calcium sparks. Science 270:633–637

    Article  PubMed  CAS  Google Scholar 

  35. Nelson MT, Quayle JM (1995) Physiological roles and properties of potassium channels in arterial smooth muscle. Am J Physiol 268:C799–C822

    PubMed  CAS  Google Scholar 

  36. Ramsey J, Austin C, Wray S (1994) Differential effects of external pH alteration on intracellular pH on intracellular pH in rat coronary and cardiac myocytes. Pflügers Arch 428:674–676

    Article  PubMed  CAS  Google Scholar 

  37. Robertson BE, Nelson MT (1994) Aminopyridine-inhibition and voltage-dependence of K+ currents in smooth muscle cells from rabbit basilar arteries. Am J Physiol 267:C1589–C1597

    PubMed  CAS  Google Scholar 

  38. Siskind MS, McCOY CE, Chobanian A, Schwartz JH (1989) Regulation of intracellular calcium by pH in vascular smooth muscle cells. Am J Physiol 256:C234–C240

    PubMed  CAS  Google Scholar 

  39. Smith GL, Austin C, Crichton C, Wray S (1998) A review of the actions and control of intracellular pH in vascular smooth muscle. Cardiovasc Res 38:316–331

    Article  PubMed  CAS  Google Scholar 

  40. Somlyo AP, Somlyo AV (1994) Signal transduction and regulation in smooth muscle. Nature 372:231–236

    Article  PubMed  CAS  Google Scholar 

  41. Song Y, Simard JM (1995) β-Adrenoceptor stimulation activates large-conductance Ca2+-activated K+ channels in smooth muscle cells from basilar artery of guinea pig. Pflügers Arch 430(6):984–993

    Article  PubMed  CAS  Google Scholar 

  42. Speake T, Elliott AC (1998) Modulation of calcium signals by intracellular pH in isolated rat pancreatic acinar cells. J Physiol 506:415–430

    Article  PubMed  CAS  Google Scholar 

  43. Stout MA (1991) Calcium transport by sarcoplasmic reticulum of vascular smooth muscle: I. MgATP-dependent and MgATP-independent calcium uptake. J Cell Physiol 149(3):383–395

    Article  PubMed  CAS  Google Scholar 

  44. Wu C, Fry CH (1998) The effects of extracellular and intracellular pH on intracellular Ca2+ regulation in guinea-pig detrusor smooth muscle. J Physiol 508:131–143

    Article  PubMed  CAS  Google Scholar 

  45. Yeon DS, Kim JS, Ahn DS, Kwon SC, Kang BS, Morgan KG, Lee YH (2002) Role of protein kinase C- or RhoA-induced Ca2+ sensitization in stretch-induced myogenic tone. Cardiovasc Res 53(2):431–438

    Article  PubMed  CAS  Google Scholar 

  46. Yodozawa S, Speake T, Elliot A (1997) Intracellular alkalinization mobilizes calcium from agonist-sensitive pools in rat lacrimal acinar cells. J Physiol 499:601–611

    PubMed  CAS  Google Scholar 

Download references

Acknowledgment

Jong Kook Park and Young Chul Kim contributed equally to this work.

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Authors and Affiliations

  1. Department of Anesthesiology and Pain Medicine, College of Medicine, Cheju National University, 66 Jejudaehakno, Jeju, Jeju Special Self-Governing Province, 690-756, South Korea

    Jong Kook Park

  2. Department of Physiology, College of Medicine, Chungbuk National University, 12 Gaeshin-dong, Hungduk-gu, Cheongju, Chungbuk, 361-804, South Korea

    Young Chul Kim & Sang Jin Lee

  3. Department of Ophtalmology, College of Medicine, Chungbuk National University, 12 Gaeshin-dong, Hungduk-gu, Cheongju, Chungbuk, 361-804, South Korea

    Mi Young Choi

  4. Department of Pharmacology, College of Medicine, Chungbuk National University, 12 Gaeshin-dong, Hungduk-gu, Cheongju, Chungbuk, 361-804, South Korea

    Woong Choi

  5. Department of Internal Medicine, College of Medicine, Chungbuk National University, 12 Gaeshin-dong, Hungduk-gu, Cheongju, Chungbuk, 361-804, South Korea

    Kyung-Kuk Hwang & Myeong-Chan Cho

  6. Anesthesiology and Pain Medicine, College of Medicine, Chungbuk National University, 12 Gaeshin-dong, Hungduk-gu, Cheongju, Chungbuk, 361-804, South Korea

    Seung Woon Lim

  7. Department of Physiology and Biophysics, Seoul National University, College of Medicine, 28 Yongon-Dong, Chongno-Gu, Seoul, 110-799, South Korea

    Jae Hoon Sim & Ki Whan Kim

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  1. Jong Kook Park
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Correspondence to Young Chul Kim.

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Park, J.K., Kim, Y.C., Sim, J.H. et al. Regulation of membrane excitability by intracellular pH (pHi) changers through Ca2+-activated K+ current (BK channel) in single smooth muscle cells from rabbit basilar artery. Pflugers Arch - Eur J Physiol 454, 307–319 (2007). https://doi.org/10.1007/s00424-007-0204-8

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