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Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. Jun 1, 1998; 101(11): 2319–2330.
PMCID: PMC508821

Molecular identification of the role of voltage-gated K+ channels, Kv1.5 and Kv2.1, in hypoxic pulmonary vasoconstriction and control of resting membrane potential in rat pulmonary artery myocytes.

Abstract

Hypoxia initiates pulmonary vasoconstriction (HPV) by inhibiting one or more voltage-gated potassium channels (Kv) in the pulmonary artery smooth muscle cells (PASMCs) of resistance arteries. The resulting membrane depolarization increases opening of voltage-gated calcium channels, raising cytosolic Ca2+ and initiating HPV. There are presently nine families of Kv channels known and pharmacological inhibitors lack the specificity to distinguish those involved in control of resting membrane potential (Em) or HPV. However, the Kv channels involved in Em and HPV have characteristic electrophysiological and pharmacological properties which suggest their molecular identity. They are slowly inactivating, delayed rectifier currents, inhibited by 4-aminopyridine (4-AP) but insensitive to charybdotoxin. Candidate Kv channels with these traits (Kv1.5 and Kv2.1) were studied. Antibodies were used to immunolocalize and functionally characterize the contribution of Kv1. 5 and Kv2.1 to PASMC electrophysiology and vascular tone. Immunoblotting confirmed the presence of Kv1.1, 1.2, 1.3, 1.5, 1.6, and 2.1, but not Kv1.4, in PASMCs. Intracellular administration of anti-Kv2.1 inhibited whole cell K+ current (IK) and depolarized Em. Anti-Kv2.1 also elevated resting tension and diminished 4-AP-induced vasoconstriction in membrane-permeabilized pulmonary artery rings. Anti-Kv1.5 inhibited IK and selectively reduced the rise in [Ca2+]i and constriction caused by hypoxia and 4-AP. However, anti-Kv1.5 neither caused depolarization nor elevated basal pulmonary artery tone. This study demonstrates that antibodies can be used to dissect the whole cell K+ currents in mammalian cells. We conclude that Kv2. 1 is an important determinant of resting Em in PASMCs from resistance arteries. Both Kv2.1 and Kv1.5 contribute to the initiation of HPV.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Kato M, Staub NC. Response of small pulmonary arteries to unilobar hypoxia and hypercapnia. Circ Res. 1966 Aug;19(2):426–440. [PubMed]
  • Vadula MS, Kleinman JG, Madden JA. Effect of hypoxia and norepinephrine on cytoplasmic free Ca2+ in pulmonary and cerebral arterial myocytes. Am J Physiol. 1993 Dec;265(6 Pt 1):L591–L597. [PubMed]
  • Madden JA, Vadula MS, Kurup VP. Effects of hypoxia and other vasoactive agents on pulmonary and cerebral artery smooth muscle cells. Am J Physiol. 1992 Sep;263(3 Pt 1):L384–L393. [PubMed]
  • Post JM, Hume JR, Archer SL, Weir EK. Direct role for potassium channel inhibition in hypoxic pulmonary vasoconstriction. Am J Physiol. 1992 Apr;262(4 Pt 1):C882–C890. [PubMed]
  • Yuan XJ, Goldman WF, Tod ML, Rubin LJ, Blaustein MP. Hypoxia reduces potassium currents in cultured rat pulmonary but not mesenteric arterial myocytes. Am J Physiol. 1993 Feb;264(2 Pt 1):L116–L123. [PubMed]
  • Archer SL, Huang JM, Reeve HL, Hampl V, Tolarová S, Michelakis E, Weir EK. Differential distribution of electrophysiologically distinct myocytes in conduit and resistance arteries determines their response to nitric oxide and hypoxia. Circ Res. 1996 Mar;78(3):431–442. [PubMed]
  • McMurtry IF, Davidson AB, Reeves JT, Grover RF. Inhibition of hypoxic pulmonary vasoconstriction by calcium antagonists in isolated rat lungs. Circ Res. 1976 Feb;38(2):99–104. [PubMed]
  • Mohazzab KM, Fayngersh RP, Kaminski PM, Wolin MS. Potential role of NADH oxidoreductase-derived reactive O2 species in calf pulmonary arterial PO2-elicited responses. Am J Physiol. 1995 Nov;269(5 Pt 1):L637–L644. [PubMed]
  • Chandy KG, Gutman GA. Nomenclature for mammalian potassium channel genes. Trends Pharmacol Sci. 1993 Dec;14(12):434–434. [PubMed]
  • Grissmer S, Nguyen AN, Aiyar J, Hanson DC, Mather RJ, Gutman GA, Karmilowicz MJ, Auperin DD, Chandy KG. Pharmacological characterization of five cloned voltage-gated K+ channels, types Kv1.1, 1.2, 1.3, 1.5, and 3.1, stably expressed in mammalian cell lines. Mol Pharmacol. 1994 Jun;45(6):1227–1234. [PubMed]
  • Yuan XJ, Goldman WF, Tod ML, Rubin LJ, Blaustein MP. Ionic currents in rat pulmonary and mesenteric arterial myocytes in primary culture and subculture. Am J Physiol. 1993 Feb;264(2 Pt 1):L107–L115. [PubMed]
  • Post JM, Gelband CH, Hume JR. [Ca2+]i inhibition of K+ channels in canine pulmonary artery. Novel mechanism for hypoxia-induced membrane depolarization. Circ Res. 1995 Jul;77(1):131–139. [PubMed]
  • Yuan XJ. Voltage-gated K+ currents regulate resting membrane potential and [Ca2+]i in pulmonary arterial myocytes. Circ Res. 1995 Aug;77(2):370–378. [PubMed]
  • Smirnov SV, Robertson TP, Ward JP, Aaronson PI. Chronic hypoxia is associated with reduced delayed rectifier K+ current in rat pulmonary artery muscle cells. Am J Physiol. 1994 Jan;266(1 Pt 2):H365–H370. [PubMed]
  • Osipenko ON, Evans AM, Gurney AM. Regulation of the resting potential of rabbit pulmonary artery myocytes by a low threshold, O2-sensing potassium current. Br J Pharmacol. 1997 Apr;120(8):1461–1470. [PMC free article] [PubMed]
  • Kubo Y, Reuveny E, Slesinger PA, Jan YN, Jan LY. Primary structure and functional expression of a rat G-protein-coupled muscarinic potassium channel. Nature. 1993 Aug 26;364(6440):802–806. [PubMed]
  • Ferguson JJ. American College of Cardiology 45th Annual Scientific Session, Orlando, Florida, March 24 to 27, 1996. Circulation. 1996 Jul 1;94(1):1–5. [PubMed]
  • Bouchard R, Fedida D. Closed- and open-state binding of 4-aminopyridine to the cloned human potassium channel Kv1.5. J Pharmacol Exp Ther. 1995 Nov;275(2):864–876. [PubMed]
  • Albrecht B, Lorra C, Stocker M, Pongs O. Cloning and characterization of a human delayed rectifier potassium channel gene. Receptors Channels. 1993;1(2):99–110. [PubMed]
  • Yang T, Snyders DJ, Roden DM. Inhibition of cardiac potassium currents by the vesnarinone analog OPC-18790: comparison with quinidine and dofetilide. J Pharmacol Exp Ther. 1997 Mar;280(3):1170–1175. [PubMed]
  • Garcia ML, Garcia-Calvo M, Hidalgo P, Lee A, MacKinnon R. Purification and characterization of three inhibitors of voltage-dependent K+ channels from Leiurus quinquestriatus var. hebraeus venom. Biochemistry. 1994 Jun 7;33(22):6834–6839. [PubMed]
  • Taglialatela M, Vandongen AM, Drewe JA, Joho RH, Brown AM, Kirsch GE. Patterns of internal and external tetraethylammonium block in four homologous K+ channels. Mol Pharmacol. 1991 Aug;40(2):299–307. [PubMed]
  • Shi G, Kleinklaus AK, Marrion NV, Trimmer JS. Properties of Kv2.1 K+ channels expressed in transfected mammalian cells. J Biol Chem. 1994 Sep 16;269(37):23204–23211. [PubMed]
  • Trimmer JS. Expression of Kv2.1 delayed rectifier K+ channel isoforms in the developing rat brain. FEBS Lett. 1993 Jun 14;324(2):205–210. [PubMed]
  • Patel AJ, Lazdunski M, Honoré E. Kv2.1/Kv9.3, a novel ATP-dependent delayed-rectifier K+ channel in oxygen-sensitive pulmonary artery myocytes. EMBO J. 1997 Nov 17;16(22):6615–6625. [PMC free article] [PubMed]
  • Wang J, Juhaszova M, Rubin LJ, Yuan XJ. Hypoxia inhibits gene expression of voltage-gated K+ channel alpha subunits in pulmonary artery smooth muscle cells. J Clin Invest. 1997 Nov 1;100(9):2347–2353. [PMC free article] [PubMed]
  • Adda S, Fleischmann BK, Freedman BD, Yu M, Hay DW, Kotlikoff MI. Expression and function of voltage-dependent potassium channel genes in human airway smooth muscle. J Biol Chem. 1996 May 31;271(22):13239–13243. [PubMed]
  • Tristani-Firouzi M, Reeve HL, Tolarova S, Weir EK, Archer SL. Oxygen-induced constriction of rabbit ductus arteriosus occurs via inhibition of a 4-aminopyridine-, voltage-sensitive potassium channel. J Clin Invest. 1996 Nov 1;98(9):1959–1965. [PMC free article] [PubMed]
  • Rhodes KJ, Keilbaugh SA, Barrezueta NX, Lopez KL, Trimmer JS. Association and colocalization of K+ channel alpha- and beta-subunit polypeptides in rat brain. J Neurosci. 1995 Jul;15(7 Pt 2):5360–5371. [PubMed]
  • Scott VE, Muniz ZM, Sewing S, Lichtinghagen R, Parcej DN, Pongs O, Dolly JO. Antibodies specific for distinct Kv subunits unveil a heterooligomeric basis for subtypes of alpha-dendrotoxin-sensitive K+ channels in bovine brain. Biochemistry. 1994 Feb 22;33(7):1617–1623. [PubMed]
  • Sheng M, Tsaur ML, Jan YN, Jan LY. Contrasting subcellular localization of the Kv1.2 K+ channel subunit in different neurons of rat brain. J Neurosci. 1994 Apr;14(4):2408–2417. [PubMed]
  • Takimoto K, Fomina AF, Gealy R, Trimmer JS, Levitan ES. Dexamethasone rapidly induces Kv1.5 K+ channel gene transcription and expression in clonal pituitary cells. Neuron. 1993 Aug;11(2):359–369. [PubMed]
  • Bouthenet ML, Souil E, Martres MP, Sokoloff P, Giros B, Schwartz JC. Localization of dopamine D3 receptor mRNA in the rat brain using in situ hybridization histochemistry: comparison with dopamine D2 receptor mRNA. Brain Res. 1991 Nov 15;564(2):203–219. [PubMed]
  • Cuevas EC, Bateman AC, Wilkins BS, Johnson PA, Williams JH, Lee AH, Jones DB, Wright DH. Microwave antigen retrieval in immunocytochemistry: a study of 80 antibodies. J Clin Pathol. 1994 May;47(5):448–452. [PMC free article] [PubMed]
  • Archer SL, Nelson DP, Weir EK. Simultaneous measurement of O2 radicals and pulmonary vascular reactivity in rat lung. J Appl Physiol (1985) 1989 Nov;67(5):1903–1911. [PubMed]
  • Archer SL, Cowan NJ. Measurement of endothelial cytosolic calcium concentration and nitric oxide production reveals discrete mechanisms of endothelium-dependent pulmonary vasodilatation. Circ Res. 1991 Jun;68(6):1569–1581. [PubMed]
  • Bekele-Arcuri Z, Matos MF, Manganas L, Strassle BW, Monaghan MM, Rhodes KJ, Trimmer JS. Generation and characterization of subtype-specific monoclonal antibodies to K+ channel alpha- and beta-subunit polypeptides. Neuropharmacology. 1996;35(7):851–865. [PubMed]
  • Archer SL, Huang J, Henry T, Peterson D, Weir EK. A redox-based O2 sensor in rat pulmonary vasculature. Circ Res. 1993 Dec;73(6):1100–1112. [PubMed]
  • López-Barneo J, López-López JR, Ureña J, González C. Chemotransduction in the carotid body: K+ current modulated by PO2 in type I chemoreceptor cells. Science. 1988 Jul 29;241(4865):580–582. [PubMed]
  • Youngson C, Nurse C, Yeger H, Cutz E. Oxygen sensing in airway chemoreceptors. Nature. 1993 Sep 9;365(6442):153–155. [PubMed]
  • Weir EK, Reeve HL, Huang JM, Michelakis E, Nelson DP, Hampl V, Archer SL. Anorexic agents aminorex, fenfluramine, and dexfenfluramine inhibit potassium current in rat pulmonary vascular smooth muscle and cause pulmonary vasoconstriction. Circulation. 1996 Nov 1;94(9):2216–2220. [PubMed]
  • Overturf KE, Russell SN, Carl A, Vogalis F, Hart PJ, Hume JR, Sanders KM, Horowitz B. Cloning and characterization of a Kv1.5 delayed rectifier K+ channel from vascular and visceral smooth muscles. Am J Physiol. 1994 Nov;267(5 Pt 1):C1231–C1238. [PubMed]
  • Duprat F, Guillemare E, Romey G, Fink M, Lesage F, Lazdunski M, Honore E. Susceptibility of cloned K+ channels to reactive oxygen species. Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11796–11800. [PMC free article] [PubMed]
  • Mays DJ, Foose JM, Philipson LH, Tamkun MM. Localization of the Kv1.5 K+ channel protein in explanted cardiac tissue. J Clin Invest. 1995 Jul;96(1):282–292. [PMC free article] [PubMed]
  • Morales MJ, Wee JO, Wang S, Strauss HC, Rasmusson RL. The N-terminal domain of a K+ channel beta subunit increases the rate of C-type inactivation from the cytoplasmic side of the channel. Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15119–15123. [PMC free article] [PubMed]
  • Lee TE, Philipson LH, Kuznetsov A, Nelson DJ. Structural determinant for assembly of mammalian K+ channels. Biophys J. 1994 Mar;66(3 Pt 1):667–673. [PMC free article] [PubMed]

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