Skip to main content

Advertisement

SpringerLink
Log in

ATP-sensitive K+ channels (Kir6.1/SUR1) regulate gap junctional coupling in cochlear-supporting cells

  • Ion channels, receptors and transporters
  • Published:
Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

Using the double whole-cell patch-clamp technique, we found that the absence of intracellular ATP led to gap junction uncoupling in cochlear-supporting Hensen cells. The uncoupling was observed as a progressive reduction of the gap junctional electrical conductance from a starting value of approximately 40 nS to less than 0.04 nS within 10–20 min. The conductance rundown was partly avoided by at least 3 mM ATP and completely suppressed by 5 mM ATP or 5ʹ-adenylyl-imidodiphosphate (AMP-PNP), the non-hydrolysable ATP analog, in the pipette filling solution, suggesting that ATP was needed as ligand and not as a hydrolysable energy supplier or substrate for enzymatic reactions. The effect of intracellular ATP was mimicked by the external application of barium, a nonselective blocker of inwardly rectifying K+ (Kir) channels, and glibenclamide, an inhibitor of the ATP-sensitive Kir channels (KATP). Moreover a Ba2+-sensitive whole-cell inward current was observed in absence of internal ATP. We propose that the internal ATP kept the KATP channels in a closed state, thereby maintaining the gap junction coupling of Hensen cells. The immunostaining of guinea pig cochlear tissue revealed for the first time the expression of the KATP channel subunits Kir6.1 and SUR1 in Hensen cells and supported the proposed hypothesis. The results suggest that KATP channels, as regulator of the gap junction coupling in Hensen cells, could be the physiological link between the metabolic state of the supporting cells and K+ recycling in the organ of Corti.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Blödow A, Ngezahayo A, Ernst A, Kolb HA (2003) Calmodulin antagonists suppress gap junctional coupling in isolated Hensen cells of the guinea pig cochlea. Pflugers Arch 446:36–41

    Article  PubMed  Google Scholar 

  2. Bruzzone R, White TW, Paul DL (1996) Connections with connexins: the molecular basis of direct intercellular signalling. Eur J Biochem 244:1–27

    Article  Google Scholar 

  3. Chen J, Zhao HB (2014) The role of an inwardly rectifying K+ channel (Kir4.1) in the inner ear and hearing loss. Neuroscience 0:137–146

    PubMed Central  Google Scholar 

  4. Cohen-Salmon M, Ott T, Michel V, Hardelin JP, Perfettini I, Eybalin M, Wu T, Marcus DC, Wangemann P, Willecke K, Petit C (2002) Targeted ablation of connexin26 in the inner ear epithelium gpa junction network causes hearing impairment and cell death. Curr Biol 12:1006–1111

    Article  Google Scholar 

  5. Donaldson PJ, Dong Y, Roos M, Green C, Goodenough DA, Kistler J (1995) Changes in lens connexin expression lead to increased gap junctional voltage dependence and conductance. Am J Physiol 69:C590–C600

    Google Scholar 

  6. Evans HW, Martin PE (2002) Gap junctions: structure and function (review). Mol Membr Biol 19:121–136

    Article  CAS  PubMed  Google Scholar 

  7. Flock A, Flock B, Fridberger A, Scarfone E, Ulfendahl M (1999) Gap junctions and connexin expression in the inner ear. Novartis Found Symp 219:134–136

    Google Scholar 

  8. Forge A, Becker D, Casalotti S, Edwards J, Marziano N, Nickel R (2002) Connexins and gap junctions in the inner ear. Audiol Neurootol 7:141–145

    Article  CAS  PubMed  Google Scholar 

  9. Gale JE, Jagger DJ (2010) Cochlear supporting cells. In: Fuchs PA (ed) The Oxford handbook of auditory science: the ear. Oxford UP, Oxford, pp 307–327

    Google Scholar 

  10. Granda B, Tabernero A, Sanchez-Abarca LI, Medina JM (1998) The K-ATP channel regulates the effect of Ca2+ on gap junction permeability in cultured astrocytes. FEBS Lett 427:41–45

    Article  CAS  PubMed  Google Scholar 

  11. Grifa A, Wagner CA, D’Ambrosio L, Melchionda S, Bernardi F, Lopez-Bigas N, Rabionet R, Arbones M, Monica MD, Estivill X, Zelante L, Lang F, Gasparini P (1999) Mutations in GJB6 cause nonsyndromic autosomal dominant deafness at DFNA3 locus. Nat Genet 23:16–18

    CAS  PubMed  Google Scholar 

  12. Hibino H, Horio Y, Ianobe A, Doi K, Ito M, Yamada M, Gotow T, Uchiyama Y, Kawamura M, Kubo T, Kurachi Y (1997) An ATP-dependent inwardly rectifying potassium channel, KAB-2 (Kir4.1), in cochlear stria vascularis of inner ear: its specific subcellular localization and correlation with the formation of endocochlear potential. J Neurosci 17:4711–4721

    CAS  PubMed  Google Scholar 

  13. Hibino H, Inanobe A, Furutani K, Murakami S, Findlay I, Kurachi Y (2010) Inwardly rectifying potassium channels: their structure, function, and physiological roles. Physiol Rev 90:291–366

    Article  CAS  PubMed  Google Scholar 

  14. Jiang K, Wang J, Zhao C, Feng M, Shen Z, Yu Z, Xia Z (2011) Regulation of gap junctional communication by astrocytic mitochondrial K(ATP) channels following neurotoxin administration in in vitro and in vivo models. Neurosignals 19:63–74

    Article  CAS  PubMed  Google Scholar 

  15. Kelsell DP, Dunlop J, Stevens HP, Lench NJ, Liang JN, Parry G, Mueller RF, Leigh IM (1997) Connexin 26 mutations in heredirary nonsyndromic sensorineural deafness. Nature 387:80–83

    Article  CAS  PubMed  Google Scholar 

  16. Kikuchi T, Adams JC, Miyabe Y, So E, Kobayashi (2000) Potassium ion recycling pathway via gap junction systems in the mammalian cochlea and its interruption in hereditary nonsyndromic deafness. Med Electron Microsc 33:51–56

    Article  CAS  PubMed  Google Scholar 

  17. Kitano I, Mori N, Matsunaga T (1995) Role of ATP-sensitive K+ channels in anoxia-sensitive negative potential of endolymph. Hear Res 90:24–30

    Article  CAS  PubMed  Google Scholar 

  18. Kumar NM, Gilula NB (1996) The gap junction communication channel. Cell 84:381–388

    Article  CAS  PubMed  Google Scholar 

  19. Lautermann J, ten Cate WJF, Altenhoff P, Gruemmer R, Traub O, Frank HG, Jahnke K, Winterhager E (1998) Expression of the gap-junction connexins 26 and 30 in the rat cochlea. Cell Tissue Res 294:415–420

    Article  CAS  PubMed  Google Scholar 

  20. Marcus DC, Wu T, Wangemann P, Kofuji P (2002) KCNJ10 (Kir4.1) potassium channel knockout abolishes endocochlear potential. Am J Physiol 282:C403–C407

    Article  CAS  Google Scholar 

  21. Naitoh K, Ichikawa Y, Miura T, Nakamura Y, Miki T, Ikeda Y, Kobayashi H, Nishihara M, Ohori K, Shimamoto K (2006) MitoKATP channel activation suppresses gap junction permeability in the ischemic myocardium by an ERK-dependent mechanism. Cardiovasc Res 70:374–383

    Article  CAS  PubMed  Google Scholar 

  22. Nenov AP, Chen C, Bobbin RP (1998) Outward rectifying potassium currents are the dominant voltage activated currents present in Deiters’ cells. Hear Res 123:168–182

    Article  CAS  PubMed  Google Scholar 

  23. Neyton J, Trautmann A (1985) Single-channel currents of an intercellular junction. Nature 317:331–335

    Article  CAS  PubMed  Google Scholar 

  24. Ngezahayo A, Altmann B, Kolb HA (2003) Regulation of ion fluxes, cell volume and gap junctional coupling by cGMP in GFSHR-17 granulosa cells. J Membrane Biol 194:165–176

    Article  CAS  Google Scholar 

  25. Nichols CG (2006) KATP channels as molecular sensors of cellular metabolism. Nature 440:470–476

    Article  CAS  PubMed  Google Scholar 

  26. Rocheleau JV, Remedi MS, Granada B, Head WS, Koster JC, Nichols CG, Piston DW (2006) Critical role of gap junction coupled KATP channel activity for regulated insulin secretion. PLoS Biol 4(2):e26, Epub 2006 Jan 17

  27. Saez JC, Spray DC, Nairn AC, Hertzberg E, Greengard P, Bennett MVL (1986) cAMP increases junctional conductance and stimulates phosphorylation of the 27-kDa principal gap junction polypeptide. Proc Natl Acad Sci 83:2473–2477

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Santos-Sacchi J (1985) The effects of cytoplasmic acidification upon electrical coupling in the organ of Corti. Hear Res 19:207–215

    Article  CAS  PubMed  Google Scholar 

  29. Santos-Sacchi J (1986) The temperature dependence of electrical coupling in the organ of Corti. Hear Res 21:205–211

    Article  CAS  PubMed  Google Scholar 

  30. Santos-Sacchi J (2000) Cell coupling in the organ of Corti. Brain Res Rev 32:167–171

    Article  CAS  PubMed  Google Scholar 

  31. Sato Y, Santos-Sacchi J (1994) Cell coupling in the supporting cells of Corti’s organ: sensitivity to intracellular H+ and Ca++. Hear Res 80:21–24

    Article  CAS  PubMed  Google Scholar 

  32. Söhl G, Willecke K (2004) Gap junctions and the connexin protein family. Cardiovasc Res 62:228–232

    Article  PubMed  Google Scholar 

  33. Solan JL, Lampe PD (2005) Connexin phosphorylation as a regulatory event linked to gap junction channel assembly. Biochim Biophys Acta 1711:154–163

    Article  CAS  PubMed  Google Scholar 

  34. Spicer SS, Schulte BA (1996) The fine structure of spiral ligament cells relates to ion return to the stria and varies with place-frequency. Hear Res 100:80–100

    Article  CAS  PubMed  Google Scholar 

  35. Spitzer N, Sammons GS, Price EM (2011) Autofluorescent cells in rat brain can be convincing impostors in green fluorescent reporter studies. J Neurosci Methods 197:48–55

    Article  PubMed  PubMed Central  Google Scholar 

  36. Todt I, Ngezahayo A, Ernst A, Kolb HA (1999) Inhibition of gap junctional coupling in cochlear supporting cells by gentamicin. Pflugers Arch 438:865–867

    Article  CAS  PubMed  Google Scholar 

  37. Todt I, Ngezahayo A, Ernst A, Kolb HA (2001) Hydrogenperoxide inhibits gap junctional coupling and modulates intracellular free calcium in cochlear Hensen-cells. J Membr Biol 181:107–114

    Article  CAS  PubMed  Google Scholar 

  38. Velasco A, Tabernero A, Granda B, Medina JM (2000) ATP-sensitive potassium channel regulates astrocytic gap junction permeability by a Ca2+-independent mechanism. J Neurochem 74:1249–1256

    Article  CAS  PubMed  Google Scholar 

  39. Vera B, Sanchez-Abarca LI, Bolanos JP, Medina JM (1996) Inhibition of astrocyte gap junctional communication by ATP depletion is revered by calcium sequestration. FEBS 392:225–228

    Article  CAS  Google Scholar 

  40. Willecke K, Eiberger J, Degen J, Eckhardt D, Romualdi A, Güldenagel M, Deutsch U, Söhl U (2002) Structural and functional diversity of connexin genes in the mouse and human genome. Biol Chem 383:725–737

    Article  CAS  PubMed  Google Scholar 

  41. Wollbold J, Jaster R, Müller S, Rateitschak K, Wolkenhauer O (2014) Anti-inflammatory effects of reactive oxygen species—a multi-valued logical model validated by formal concept analysis. BMC Syst Biol 8:101

    Article  PubMed  PubMed Central  Google Scholar 

  42. Xia JH, Liu CY, Tang BS, Pan Q, Huang L, Dai HP, Zhang BR, Xie W, Hu DX, Zheng D, Shi XL, Wang DA, Xia K, Yu KP, Liao XD, Feng Y, Yang YF, Xiao JY, Xie DH, Huang JZ (1998) Mutations in the gene encoding gap junction protein beta-3 associated with autosomal dominant hearing impairment. Nat Genet 20:370–373

    Article  CAS  PubMed  Google Scholar 

  43. Yang J, Wang J (2002) Possible function of outward potassium currents in isolated Deiters’cells of guinea pig cochlea. Chin Med J 115:264–267

    CAS  PubMed  Google Scholar 

  44. Zelante L, Gasparini P, Estivill X, Melchionda S, D’Agruma L, Govea N, Milá M, Monica MD, Lutfi J, Shohat M, Mansfield E, Delgrosso K, Rappaport E, Surrey S, Fortina P (1997) Connexin26 mutations associated with the most common form of non-syndromic neurosensory autosomal recessive deafness (DFNB1) in Mediterraneans. Hum Mol Genet 6:1605–1609

    Article  CAS  PubMed  Google Scholar 

  45. Zhao HB, Santos-Sacchi J (1998) Effect of membrane tension on gap junctional conductance of supporting cells in Corti’s organ. J Gen Physiol 112:447–455

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Zhao HB, Yu N, Fleming CR (2005) Gap junctional hemichannel-mediated ATP release and hearing controls in the inner ear. Proc Natl Acad Sci U S A 102:18724–18729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Zhao HB, Kikuchi T, Ngezahayo A, White TW (2006) Gap junction and cochlear homeostasis. J Membr Biol 209:177–186

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

The authors thank Ina G. Siller for the help with imaging the cochlear slides. We thank Dr. Kathrin Rübensam, ZTL, MH Hannover, and Dr. Henning Vogt, HNO-Klinik, MH Hannover for their kind help with the animals.

Author information

Authors and Affiliations

  1. Department of Otolaryngology, Head and Neck Surgery and Communication Disorders, HELIOS-Klinikum Pirna, Pirna, Germany

    Alexander Blödow

  2. Institute of Biophysics, Leibniz University Hannover, Herrenhäuserstr. 2, 30419, Hannover, Germany

    Daniela Begandt, Almke Bader, Annegret Becker & Anaclet Ngezahayo

  3. Institute of Audioneurotechnology and Department of Experimental Otology, ENT Clinics, Hannover Medical University, Hannover, Germany

    Alice Burghard, Daniela Kühne & Andrej Kral

  4. Center for Systems Neuroscience, Hannover, Germany

    Andrej Kral & Anaclet Ngezahayo

Authors
  1. Alexander Blödow
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniela Begandt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Almke Bader
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Annegret Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alice Burghard
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daniela Kühne
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Andrej Kral
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Anaclet Ngezahayo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anaclet Ngezahayo.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Blödow, A., Begandt, D., Bader, A. et al. ATP-sensitive K+ channels (Kir6.1/SUR1) regulate gap junctional coupling in cochlear-supporting cells. Pflugers Arch - Eur J Physiol 468, 1215–1222 (2016). https://doi.org/10.1007/s00424-016-1815-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-016-1815-8

Keywords

Search

Navigation