Pflügers Archiv - European Journal of Physiology

, Volume 465, Issue 2, pp 233–245 | Cite as

Downregulation of the renal outer medullary K+ channel ROMK by the AMP-activated protein kinase

  • Balasaheb Siraskar
  • Dan Yang Huang
  • Tatsiana Pakladok
  • Gulab Siraskar
  • Mentor Sopjani
  • Ioana Alesutan
  • Yulia Kucherenko
  • Ahmad Almilaji
  • Vasudharani Devanathan
  • Ekaterina Shumilina
  • Michael Föller
  • Carlos Munoz
  • Florian Lang
Ion Channels, Receptors and Transporters


The 5′-adenosine monophosphate-activated serine/threonine protein kinase (AMPK) is stimulated by energy depletion, increase in cytosolic Ca2+ activity, oxidative stress, and nitric oxide. AMPK participates in the regulation of the epithelial Na+ channel ENaC and the voltage-gated K+ channel KCNE1/KCNQ1. It is partially effective by decreasing PIP2 formation through the PI3K pathway. The present study explored whether AMPK regulates the renal outer medullary K+ channel ROMK. To this end, cRNA encoding ROMK was injected into Xenopus oocytes with and without additional injection of constitutively active AMPKγR70Q (AMPKα1-HA+AMPKβ1-Flag+AMPKγ1R70Q), or of inactive AMPKαK45R (AMPKα1K45R+AMPKβ1-Flag+AMPKγ1-HA), and the current determined utilizing two-electrode voltage-clamp and single channel patch clamp. ROMK protein abundance was measured utilizing chemiluminescence in Xenopus oocytes and western blot in whole kidney tissue. Moreover, renal Na+ and K+ excretion were determined in AMPKα1-deficient mice (ampk−/−) and wild-type mice (ampk+/+) prior to and following an acute K+ load (111 mM KCl, 30 mM NaHCO3, 4.7 mM NaCl, and 2.25 g/dl BSA) at a rate of 500 μl/h. As a result, coexpression of AMPKγR70Q but not of AMPKαK45R significantly decreased the current in ROMK1-expressing Xenopus oocytes. Injection of phosphatidylinositol PI(4,5)P2 significantly increased the current in ROMK1-expressing Xenopus oocytes, an effect reversed in the presence of AMPKγR70Q. Under control conditions, no significant differences between ampk−/− and ampk+/+ mice were observed in glomerular filtration rate (GFR), urinary flow rate, serum aldosterone, plasma Na+, and K+ concentrations as well as absolute and fractional Na+ and K+ excretion. Following an acute K+ load, GFR, urinary flow rate, serum aldosterone, plasma Na+, and K+ concentration were again similar in both genotypes, but renal absolute and fractional Na+ and K+ excretion were higher in ampk−/− than in ampk+/+ mice. According to micropuncture following a K+ load, delivery of Na+ to the early distal tubule but not delivery of K+ to late proximal and early distal tubules was increased in ampk−/− mice. The upregulation of renal ROMK1 protein expression by acute K+ load was more pronounced in ampk−/− than in ampk+/+ mice. In conclusion, AMPK downregulates ROMK, an effect compromising the ability of the kidney to excrete K+ following an acute K+ load.


Energy depletion K+ channels ROMK Kaliuresis Natriuresis 


  1. 1.
    Albert AP, Woollhead AM, Mace OJ, Baines DL (2008) AICAR decreases the activity of two distinct amiloride-sensitive Na+-permeable channels in H441 human lung epithelial cell monolayers. Am J Physiol Lung Cell Mol Physiol 295:L837–L848PubMedCrossRefGoogle Scholar
  2. 2.
    Alers S, Loffler AS, Wesselborg S, Stork B (2012) AMPK-mTOR-Ulk1/2 in the regulation of autophagy: crosstalk, shortcuts and feedbacks. Mol Cell Biol 32:2–11PubMedCrossRefGoogle Scholar
  3. 3.
    Alesutan I, Munoz C, Sopjani M, Dermaku-Sopjani M, Michael D, Fraser S, Kemp BE, Seebohm G, Foller M, Lang F (2011) Inhibition of Kir2.1 (KCNJ2) by the AMP-activated protein kinase. Biochem Biophys Res Commun 408:505–510PubMedCrossRefGoogle Scholar
  4. 4.
    Alesutan IS, Foller M, Sopjani M, Dermaku-Sopjani M, Zelenak C, Frohlich H, Velic A, Fraser S, Kemp BE, Seebohm G, Volkl H, Lang F (2011) Inhibition of the heterotetrameric K+ channel KCNQ1/KCNE1 by the AMP-activated protein kinase. Mol Membr Biol 28:79–89PubMedCrossRefGoogle Scholar
  5. 5.
    Almaca J, Kongsuphol P, Hieke B, Ousingsawat J, Viollet B, Schreiber R, Amaral MD, Kunzelmann K (2009) AMPK controls epithelial Na(+) channels through Nedd4-2 and causes an epithelial phenotype when mutated. Pflugers Arch 458:713–721PubMedCrossRefGoogle Scholar
  6. 6.
    Alzamora R, Gong F, Rondanino C, Lee JK, Smolak C, Pastor-Soler NM, Hallows KR (2010) AMP-activated protein kinase inhibits KCNQ1 channels through regulation of the ubiquitin ligase Nedd4-2 in renal epithelial cells. Am J Physiol Ren Physiol 299:F1308–F1319CrossRefGoogle Scholar
  7. 7.
    Becker S, Reinehr R, Graf D, vom Dahl S, Haussinger D (2007) Hydrophobic bile salts induce hepatocyte shrinkage via NADPH oxidase activation. Cell Physiol Biochem 19:89–98PubMedCrossRefGoogle Scholar
  8. 8.
    Bhalla V, Oyster NM, Fitch AC, Wijngaarden MA, Neumann D, Schlattner U, Pearce D, Hallows KR (2006) AMP-activated kinase inhibits the epithelial Na+ channel through functional regulation of the ubiquitin ligase Nedd4-2. J Biol Chem 281:26159–26169PubMedCrossRefGoogle Scholar
  9. 9.
    Bohmer C, Sopjani M, Klaus F, Lindner R, Laufer J, Jeyaraj S, Lang F, Palmada M (2010) The serum and glucocorticoid inducible kinases SGK1-3 stimulate the neutral amino acid transporter SLC6A19. Cell Physiol Biochem 25:723–732PubMedCrossRefGoogle Scholar
  10. 10.
    Boiteux A, Hess B (1981) Design of glycolysis. Philos Trans R Soc Lond B Biol Sci 293:5–22PubMedCrossRefGoogle Scholar
  11. 11.
    Bortner CD, Cidlowski JA (2004) The role of apoptotic volume decrease and ionic homeostasis in the activation and repression of apoptosis. Pflugers Arch 448:313–318PubMedCrossRefGoogle Scholar
  12. 12.
    Boulpaep E (2009) Protein-protein interactions among ion channels regulate ion transport in the kidney. Bull Mem Acad R Med Belg 164:133–141PubMedGoogle Scholar
  13. 13.
    Carattino MD, Edinger RS, Grieser HJ, Wise R, Neumann D, Schlattner U, Johnson JP, Kleyman TR, Hallows KR (2005) Epithelial sodium channel inhibition by AMP-activated protein kinase in oocytes and polarized renal epithelial cells. J Biol Chem 280:17608–17616PubMedCrossRefGoogle Scholar
  14. 14.
    Eckey K, Strutz-Seebohm N, Katz G, Fuhrmann G, Henrion U, Pott L, Linke WA, Arad M, Lang F, Seebohm G (2010) Modulation of human ether a gogo related channels by CASQ2 contributes to etiology of catecholaminergic polymorphic ventricular tachycardia (CPVT). Cell Physiol Biochem 26:503–512PubMedCrossRefGoogle Scholar
  15. 15.
    Evans AM, Mustard KJ, Wyatt CN, Peers C, Dipp M, Kumar P, Kinnear NP, Hardie DG (2005) Does AMP-activated protein kinase couple inhibition of mitochondrial oxidative phosphorylation by hypoxia to calcium signaling in O2-sensing cells? J Biol Chem 280:41504–41511PubMedCrossRefGoogle Scholar
  16. 16.
    Fang L, Li D, Welling PA (2010) Hypertension resistance polymorphisms in ROMK (Kir1.1) alter channel function by different mechanisms. Am J Physiol Ren Physiol 299:F1359–F1364CrossRefGoogle Scholar
  17. 17.
    Foller M, Kasinathan RS, Duranton C, Wieder T, Huber SM, Lang F (2006) PGE2-induced apoptotic cell death in K562 human leukaemia cells. Cell Physiol Biochem 17:201–210PubMedCrossRefGoogle Scholar
  18. 18.
    Foller M, Sopjani M, Koka S, Gu S, Mahmud H, Wang K, Floride E, Schleicher E, Schulz E, Munzel T, Lang F (2009) Regulation of erythrocyte survival by AMP-activated protein kinase. FASEB J 23:1072–1080PubMedCrossRefGoogle Scholar
  19. 19.
    Fraser SA, Gimenez I, Cook N, Jennings I, Katerelos M, Katsis F, Levidiotis V, Kemp BE, Power DA (2007) Regulation of the renal-specific Na+–K+–2Cl− co-transporter NKCC2 by AMP-activated protein kinase (AMPK). Biochem J 405:85–93PubMedGoogle Scholar
  20. 20.
    Gusarova GA, Trejo HE, Dada LA, Briva A, Welch LC, Hamanaka RB, Mutlu GM, Chandel NS, Prakriya M, Sznajder JI (2011) Hypoxia leads to Na, K-ATPase downregulation via Ca(2+) release-activated Ca(2+) channels and AMPK activation. Mol Cell Biol 31:3546–3556PubMedCrossRefGoogle Scholar
  21. 21.
    Hallows KR, Kobinger GP, Wilson JM, Witters LA, Foskett JK (2003) Physiological modulation of CFTR activity by AMP-activated protein kinase in polarized T84 cells. Am J Physiol Cell Physiol 284:C1297–C1308PubMedGoogle Scholar
  22. 22.
    Hallows KR, Mount PF, Pastor-Soler NM, Power DA (2010) Role of the energy sensor AMP-activated protein kinase in renal physiology and disease. Am J Physiol Renal Physiol 298:F1067–F1077PubMedCrossRefGoogle Scholar
  23. 23.
    Hamilton SR, Yao SY, Ingram JC, Hadden DA, Ritzel MW, Gallagher MP, Henderson PJ, Cass CE, Young JD, Baldwin SA (2001) Subcellular distribution and membrane topology of the mammalian concentrative Na+-nucleoside cotransporter rCNT1. J Biol Chem 276:27981–27988PubMedCrossRefGoogle Scholar
  24. 24.
    Hardie DG (2011) AMP-activated protein kinase: an energy sensor that regulates all aspects of cell function. Genes Dev 25:1895–1908PubMedCrossRefGoogle Scholar
  25. 25.
    Hosseinzadeh Z, Bhavsar SK, Sopjani M, Alesutan I, Saxena A, Dermaku-Sopjani M, Lang F (2011) Regulation of the glutamate transporters by JAK2. Cell Physiol Biochem 28:693–702PubMedCrossRefGoogle Scholar
  26. 26.
    Huang CL, Feng S, Hilgemann DW (1998) Direct activation of inward rectifier potassium channels by PIP2 and its stabilization by Gbetagamma. Nature 391:803–806PubMedCrossRefGoogle Scholar
  27. 27.
    Huang DY, Gao H, Boini KM, Osswald H, Nurnberg B, Lang F (2010) In vivo stimulation of AMP-activated protein kinase enhanced tubuloglomerular feedback but reduced tubular sodium transport during high dietary NaCl intake. Pflugers Arch 460:187–196PubMedCrossRefGoogle Scholar
  28. 28.
    Ingwersen MS, Kristensen M, Pilegaard H, Wojtaszewski JF, Richter EA, Juel C (2011) Na, K-ATPase activity in mouse muscle is regulated by AMPK and PGC-1alpha. J Membr Biol 242:1–10PubMedCrossRefGoogle Scholar
  29. 29.
    Inoki K, Kim J, Guan KL (2012) AMPK and mTOR in cellular energy homeostasis and drug targets. Annu Rev Pharmacol Toxicol 52:381–400PubMedCrossRefGoogle Scholar
  30. 30.
    Klein H, Garneau L, Trinh NT, Prive A, Dionne F, Goupil E, Thuringer D, Parent L, Brochiero E, Sauve R (2009) Inhibition of the KCa3.1 channels by AMP-activated protein kinase in human airway epithelial cells. Am J Physiol Cell Physiol 296:C285–C295PubMedCrossRefGoogle Scholar
  31. 31.
    Lang F, Rehwald W (1992) Potassium channels in renal epithelial transport regulation. Physiol Rev 72:1–32PubMedGoogle Scholar
  32. 32.
    Lira VA, Soltow QA, Long JH, Betters JL, Sellman JE, Criswell DS (2007) Nitric oxide increases GLUT4 expression and regulates AMPK signaling in skeletal muscle. Am J Physiol Endocrinol Metab 293:E1062–E1068PubMedCrossRefGoogle Scholar
  33. 33.
    Lu M, Wang T, Yan Q, Yang X, Dong K, Knepper MA, Wang W, Giebisch G, Shull GE, Hebert SC (2002) Absence of small conductance K+ channel (SK) activity in apical membranes of thick ascending limb and cortical collecting duct in ROMK (Bartter's) knockout mice. J Biol Chem 277:37881–37887PubMedCrossRefGoogle Scholar
  34. 34.
    Lupescu A, Geiger C, Zahir N, Aberle S, Lang PA, Kramer S, Wesselborg S, Kandolf R, Foller M, Lang F, Bock CT (2009) Inhibition of Na+/H+ exchanger activity by parvovirus B19 protein NS1. Cell Physiol Biochem 23:211–220PubMedCrossRefGoogle Scholar
  35. 35.
    Mace OJ, Woollhead AM, Baines DL (2008) AICAR activates AMPK and alters PIP2 association with the epithelial sodium channel ENaC to inhibit Na+ transport in H441 lung epithelial cells. J Physiol 586:4541–4557PubMedCrossRefGoogle Scholar
  36. 36.
    McCullough LD, Zeng Z, Li H, Landree LE, McFadden J, Ronnett GV (2005) Pharmacological inhibition of AMP-activated protein kinase provides neuroprotection in stroke. J Biol Chem 280:20493–20502PubMedCrossRefGoogle Scholar
  37. 37.
    McGee SL, Hargreaves M (2008) AMPK and transcriptional regulation. Front Biosci 13:3022–3033PubMedCrossRefGoogle Scholar
  38. 38.
    Mihaylova MM, Shaw RJ (2011) The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol 13:1016–1023PubMedCrossRefGoogle Scholar
  39. 39.
    Mohamed MR, Alesutan I, Foller M, Sopjani M, Bress A, Baur M, Salama RH, Bakr MS, Mohamed MA, Blin N, Lang F, Pfister M (2010) Functional analysis of a novel I71N mutation in the GJB2 gene among Southern Egyptians causing autosomal recessive hearing loss. Cell Physiol Biochem 26:959–966PubMedCrossRefGoogle Scholar
  40. 40.
    Myerburg MM, King JD Jr, Oyster NM, Fitch AC, Magill A, Baty CJ, Watkins SC, Kolls JK, Pilewski JM, Hallows KR (2010) AMPK agonists ameliorate sodium and fluid transport and inflammation in cystic fibrosis airway epithelial cells. Am J Respir Cell Mol Biol 42:676–684PubMedCrossRefGoogle Scholar
  41. 41.
    Rotte A, Pasham V, Eichenmuller M, Bhandaru M, Foller M, Lang F (2010) Upregulation of Na+/H+ exchanger by the AMP-activated protein kinase. Biochem Biophys Res Commun 398:677–682PubMedCrossRefGoogle Scholar
  42. 42.
    Schneider J, Nicolay JP, Foller M, Wieder T, Lang F (2007) Suicidal erythrocyte death following cellular K+ loss. Cell Physiol Biochem 20:35–44PubMedCrossRefGoogle Scholar
  43. 43.
    Shimizu T, Wehner F, Okada Y (2006) Inhibition of hypertonicity-induced cation channels sensitizes HeLa cells to shrinkage-induced apoptosis. Cell Physiol Biochem 18:295–302PubMedCrossRefGoogle Scholar
  44. 44.
    Sopjani M, Bhavsar SK, Fraser S, Kemp BE, Föller M, Lang F (2010) Regulation of Na+-coupled glucose carrier SGLT1 by AMP-activated protein kinase. Mol Membr Biol 27:137–144PubMedCrossRefGoogle Scholar
  45. 45.
    Sukhodub A, Jovanovic S, Du Q, Budas G, Clelland AK, Shen M, Sakamoto K, Tian R, Jovanovic A (2007) AMP-activated protein kinase mediates preconditioning in cardiomyocytes by regulating activity and trafficking of sarcolemmal ATP-sensitive K(+) channels. J Cell Physiol 210:224–236PubMedCrossRefGoogle Scholar
  46. 46.
    Taub M, Springate JE, Cutuli F (2010) Targeting of renal proximal tubule Na, K-ATPase by salt-inducible kinase. Biochem Biophys Res Commun 393:339–344PubMedCrossRefGoogle Scholar
  47. 47.
    Towler MC, Hardie DG (2007) AMP-activated protein kinase in metabolic control and insulin signaling. Circ Res 100:328–341PubMedCrossRefGoogle Scholar
  48. 48.
    Viollet B, Andreelli F, Jorgensen SB, Perrin C, Flamez D, Mu J, Wojtaszewski JF, Schuit FC, Birnbaum M, Richter E, Burcelin R, Vaulont S (2003) Physiological role of AMP-activated protein kinase (AMPK): insights from knockout mouse models. Biochem Soc Trans 31:216–219PubMedCrossRefGoogle Scholar
  49. 49.
    Winder WW, Thomson DM (2007) Cellular energy sensing and signaling by AMP-activated protein kinase. Cell Biochem Biophys 47:332–347PubMedCrossRefGoogle Scholar
  50. 50.
    Woollhead AM, Scott JW, Hardie DG, Baines DL (2005) Phenformin and 5-aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) activation of AMP-activated protein kinase inhibits transepithelial Na+ transport across H441 lung cells. J Physiol 566:781–792PubMedCrossRefGoogle Scholar
  51. 51.
    Yun CC, Palmada M, Embark HM, Fedorenko O, Feng Y, Henke G, Setiawan I, Boehmer C, Weinman EJ, Sandrasagra S, Korbmacher C, Cohen P, Pearce D, Lang F (2002) The serum and glucocorticoid-inducible kinase SGK1 and the Na+/H+ exchange regulating factor NHERF2 synergize to stimulate the renal outer medullary K+ channel ROMK1. J Am Soc Nephrol 13:2823–2830PubMedCrossRefGoogle Scholar
  52. 52.
    Zeng WZ, Liou HH, Krishna UM, Falck JR, Huang CL (2002) Structural determinants and specificities for ROMK1-phosphoinositide interaction. Am J Physiol Renal Physiol 282:F826–F834PubMedGoogle Scholar
  53. 53.
    Zheng D, MacLean PS, Pohnert SC, Knight JB, Olson AL, Winder WW, Dohm GL (2001) Regulation of muscle GLUT-4 transcription by AMP-activated protein kinase. J Appl Physiol 91:1073–1083PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Balasaheb Siraskar
    • 1
  • Dan Yang Huang
    • 2
  • Tatsiana Pakladok
    • 1
  • Gulab Siraskar
    • 1
  • Mentor Sopjani
    • 1
    • 3
  • Ioana Alesutan
    • 1
  • Yulia Kucherenko
    • 1
  • Ahmad Almilaji
    • 1
  • Vasudharani Devanathan
    • 2
  • Ekaterina Shumilina
    • 1
  • Michael Föller
    • 1
    • 4
  • Carlos Munoz
    • 1
  • Florian Lang
    • 1
  1. 1.Department of PhysiologyUniversity of TübingenTübingenGermany
  2. 2.Department of Pharmacology and Experimental TherapyInstitute of Pharmacology and Toxicology, University of TübingenTübingenGermany
  3. 3.Faculty of MedicineUniversity of PrishtinaPrishtinaKosovo
  4. 4.Campbell Family Institute for Breast Cancer Research, Ontario Cancer InstituteUniversity Health Network (UHN)TorontoCanada

Personalised recommendations