Advertisement

Psychopharmacology

, Volume 231, Issue 9, pp 2009–2018 | Cite as

Alcohol withdrawal is associated with a downregulation of large-conductance Ca2+-activated K+ channels in rat inferior colliculus neurons

  • Prosper N’GouemoEmail author
  • Martin Morad
Original Investigation

Abstract

Rationale

Large conductance calcium-activated potassium (BKCa or KCa1.1) channels are well-known molecular targets for the action of alcohol and therefore may play an important role in the pathogenesis of alcohol withdrawal syndrome.

Objectives

We evaluate the modifications of total outward K+ currents and protein expression of BKCa channels α-subunit in inferior colliculus (IC) neurons obtained from controls and rats subjected to alcohol withdrawal associated with enhanced susceptibility to seizures.

Methods

Outward K+ currents and BKCa channel proteins were measured using the whole cell configuration of patch clamp techniques and Western blot analysis, respectively.

Results

Total outward K+ current density was significantly reduced in IC neurons at 24 and 48 h during the alcohol withdrawal period when the susceptibility to seizures was maximal and absent, respectively. The iberiotoxin-sensitive (BKCa) current density and conductance also were significantly reduced at 24 h following alcohol withdrawal. Consistent with functional data, the levels of protein expression of α-subunit associated with BKCa channels also was significantly reduced in IC neurons at 24 and 48 h following alcohol withdrawal.

Conclusions

The downregulation of BKCa channels outlasts the finite period of elevated susceptibility to alcohol withdrawal seizures. These findings indicate that BKCa channels, per se, may not be fundamentally important for the generation of alcohol withdrawal seizures.

Keywords

Alcohol withdrawal BKCa channels Iberiotoxin Outward current density Protein Seizures 

Notes

Acknowledgments

This publication was made possible by Public Health Service grants (NS047193 and AA020073 to P.N., and HL62525 to M.M.) from the National Institutes of Health (NIH), and its contents are the responsibility of the authors and do not necessary represent the official views of NIH. All the experimental procedures used in this study were in accordance with the National Research Council’s Guide for Care and Use of Laboratory Animals and approved by the Georgetown University Animal Care and Use Committee.

Conflict of interest

The authors declare no conflict of interest.

References

  1. Alger BE, Nicoll RA (1980) Epileptiform burst after hyperpolarization: calcium-dependent potential in hippocampal CA1 pyramidal cells. Science 210:1122–1124PubMedCrossRefGoogle Scholar
  2. Brenner R, Chen QH, Vilaythong A, Toney GM, Noebels JL, Aldrich RW (2005) BK channel beta4 subunit reduces dentate gyrus excitability and protects against temporal lobe seizures. Nat Neurosci 8:1752–1759PubMedCrossRefGoogle Scholar
  3. Caird D (1991) Processing in the colliculli. In: Altschuler RA, Bobbin RP, Clopton BM, Hoffman DW (eds) Neurobiology of hearing, the central auditory system. Raven, New York, pp 253–292Google Scholar
  4. Carlen PL, Gurevich N, Durand D (1982) Ethanol in low doses augments calcium-mediated mechanisms measured intracellularly in hippocampal neurons. Science 215:306–309PubMedCrossRefGoogle Scholar
  5. Carlen P, Rougier-Naquet I, Reynolds JN (1990) Alterations of neuronal calcium and potassium currents during alcohol administration and withdrawal. In Porter RJ, Mattson RH, Cramer JA, Diamond I, Schoenberg DG (eds) Alcohol and seizures: basic mechanisms and clinical concepts, Davis: Philadelphia. pp 68–78Google Scholar
  6. Chakravarty DN, Faingold CL (1998) Comparison of neuronal response patterns in the external and central nuclei of inferior colliculus during ethanol administration and ethanol withdrawal. Brain Res 783:102–108PubMedCrossRefGoogle Scholar
  7. Cowmeadow RB, Krishnan HR, Atkinson NS (2005) The slowpoke gene is necessary for rapid ethanol tolerance in Drosophila. Alcohol Clin Exp Res 29:1777–1786PubMedCrossRefGoogle Scholar
  8. Davies AG, Pierce-Shimomura JT, Kim H, VanHoven MK, Thiele TR, Bonci A, Bargmann CI, McIntire SL (2003) A central role of BK potassium channel in behavioral responses to ethanol in C. elegans. Cell 115:655–666PubMedCrossRefGoogle Scholar
  9. Deisseroth K, Bito H, Tsien RW (1996) Signaling from synapse to nucleus: postsynaptic CREB phosphorylation during multiple forms of hippocampal synaptic plasticity. Neuron 16:89–101PubMedCrossRefGoogle Scholar
  10. Diez-Sampedro A, Silverman WR, Bautista JF, Richerson GB (2006) Mechanism of increased open probability by a mutation of the BK channel. J Neurophysiol 96:1507–1516PubMedCrossRefGoogle Scholar
  11. Dopico AM, Lemos JR, Treistman SN (1996) Ethanol increases the activity of large conductance, Ca2+-activated K+ channels in isolated neurophypophysial terminals. Mol Pharmacol 49:40–48PubMedGoogle Scholar
  12. Du W, Bautista JF, Yang H, ez-Sampedro A, You SA, Wang L, Kotagal P, Luders HO, Shi J, Cui J, Richerson GB, Wang QK (2005) Calcium-sensitive potassium channelopathy in human epilepsy and paroxysmal movement disorder. Nat Genet 37:733–738PubMedCrossRefGoogle Scholar
  13. Eckardt MJ, Campbell GA, Marietta CA, Majchrowicz E, Wixon HN, Weight FF (1986) Cerebral 2-deoxyglucose uptake in rats during ethanol withdrawal and post-withdrawal. Brain Res 366:1–9PubMedCrossRefGoogle Scholar
  14. Evans MS, Li Y, Faingold CF (2000) Inferior colliculus intracellular response abnormalities in vitro associated with susceptibility to ethanol withdrawal seizures. Alcohol Clin Exp Res 24:1180–1186PubMedCrossRefGoogle Scholar
  15. Faingold CL (2008) The Majochrowicz binge alcohol protocol: an intubation technique to study alcohol dependence in rats. Curr Protoc Neurosci Chapter 9:Unit 9.28Google Scholar
  16. Faingold CL, Riaz A (1995) Ethanol withdrawal induces increased firing in the inferior colliculus neurons associated with audiogenic seizure susceptibility. Exp Neurol 132:91–98PubMedCrossRefGoogle Scholar
  17. Faingold CL, Li Y, Evans MS (2000) Decreased GABA and increase glutamate-mediated activity on inferior colliculus neurons in vitro are associated with susceptibility to ethanol withdrawal seizures. Brain Res 868:287–295PubMedCrossRefGoogle Scholar
  18. Frye GD, McCown TJ, Breese GR (1983) Characterization of susceptibility to audiogenic seizures in ethanol-dependent rats after microinjection of gamma-aminobutyric acid (GABA) agonists into the inferior colliculus, substantia nigra or medial septum. J Pharmacol Exp Ther 227:663–670PubMedCentralPubMedGoogle Scholar
  19. Ghezzi A, Pohl JB, Wang Y, Atkinson NS (2010) BK channels play a counter-adaptive role in drug tolerance and dependence. Proc Natl Acad Sci U S A 107:16360–16365PubMedCentralPubMedCrossRefGoogle Scholar
  20. Ghezzi A, Krishnan HR, Atkinson NS (2012) Susceptibility to ethanol withdrawal seizures is produced by BK channel gene expression. Addict Biol. doi: 10.1111/j.1369-1600.2012.00465.x PubMedGoogle Scholar
  21. Gu N, Vervaeke K, Storm JF (2007) BK potassium channels facilitate high-frequency firing and cause early spike frequency adaptation in rat CA1 hippocampal pyramidal cells. J Physiol 580:859–882PubMedCentralPubMedCrossRefGoogle Scholar
  22. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflüger Arch 391:85–100CrossRefGoogle Scholar
  23. Hou S, Xu R, Heinemann SH, Hoshi T (2008) Reciprocal regulation of the Ca2+ and H+ sensitivity in the SLO1 BK channel conferred by the RCK1 domain. Nat Struct Mol Biol 15:403–410PubMedCentralPubMedCrossRefGoogle Scholar
  24. Jobe PC, Picchioni AL, Chin L (1973) Role of brain 5-hydroxytryptamine in audiogenic seizures in the rat. J Pharmacol Exp Ther 184:1–10PubMedGoogle Scholar
  25. Knott TK, Dopico AM, Dayanithi G, Lemos J, Triestman (2002) Integrated channel plasticity contributes to alcohol tolerance in neurophypophysial terminals. Mol Pharmacol 62:135–142PubMedCrossRefGoogle Scholar
  26. Lancaster B, Adams PR (1986) Calcium-dependent current generating the afterhyperpolarization of hippocampal neurons. J Neurophysiol 55:1268–1282PubMedGoogle Scholar
  27. Laurido C, Candia S, Wolff D, Latorre R (1991) Proton modulation a Ca2+-activated K+ channel from skeletal muscle incorporated into planar bilayer. J Gen Physiol 98:1025–1043PubMedCrossRefGoogle Scholar
  28. Little HJ, Dolin SJ, Halsey MJ (1986) Calcium channel antagonists decrease the ethanol withdrawal syndrome. Life Sci 39:2059–2065PubMedCrossRefGoogle Scholar
  29. Loane DJ, Lima PA, Marrion NV (2007) Co-assembly of N-type Ca2+ and BK channels underlies functional coupling in rat brain. J Cell Sci 120:985–995PubMedCrossRefGoogle Scholar
  30. Martin GE, Puig SI, Pietrzykowski AZ, Zadek P, Emery P, Treistman SN (2004) Alcohol tolerance in large-conductance, calcium-activated potassium channels of CNS terminals is intrinsic and includes two components: decreased ethanol potentiation and decreased channel density. J Neurosci 24:6563–6572PubMedCrossRefGoogle Scholar
  31. Martin GE, Hendrickson LM, Penta KL, Friesen RM, Pietrzykowski AZ, Tapper AR, Treistman SN (2008) Identification of a BK channel auxiliary protein controlling molecular and behavioral tolerance to alcohol. Proc Natl Acad Sci U S A 105:17543–17548PubMedCentralPubMedCrossRefGoogle Scholar
  32. McCown TJ, Breese GR (1990) Multiple withdrawals from chronic ethanol “kindles” inferior collicular seizure activity: evidence for kindling seizures associated with alcoholism. Alcohol Clin Exp Res 14:394–399PubMedCrossRefGoogle Scholar
  33. N’Gouemo P (2011) Targeting BK (big potassium) channels in epilepsy. Expert Opin Ther Targets 15:1283–1295PubMedCentralPubMedCrossRefGoogle Scholar
  34. N’Gouemo P, Morad M (2003) Ethanol withdrawal seizure susceptibility is associated with upregulation of L- and P-type Ca2+ channels currents in rat inferior colliculus neurons. Neuropharmacology 45:429–437PubMedCrossRefGoogle Scholar
  35. N’Gouemo P, Caspary DM, Faingold CL (1996) Decreased GABA effectiveness in the inferior colliculus neurons during ethanol withdrawal in rat susceptible to audiogenic seizures. Brain Res 724:200–204PubMedCrossRefGoogle Scholar
  36. N’Gouemo P, Yasuda RP, Morad M (2006) Ethanol withdrawal is accompanied by downregulation of calcium channel alpha 1B subunit in rat inferior colliculus neurons. Brain Res 1108:216–220PubMedCrossRefGoogle Scholar
  37. N’Gouemo P, Yasuda RP, Faingold CL (2009) Protein expression of small conductance calcium-activated potassium channels is altered in inferior colliculus neurons of the genetically epilepsy-prone rat. Brain Res 1270:107–111PubMedCentralPubMedCrossRefGoogle Scholar
  38. Niesen CE, Baskys A, Carlen PL (1988) Reversed ethanol effects on potassium conductances in aged hippocampal dentate granule neurons. Brain Res 445:137–141PubMedCrossRefGoogle Scholar
  39. Pietrzykowski AZ, Martin GE, Puig SI, Knott TK, Lemos JR, Treistman SN (2004) Alcohol tolerance in large-conductance, calcium-activated potassium channels of CNS terminals is intrinsic and includes two components: decreased ethanol potentiation and decreased channel density. J Neurosci 24:8322–8332PubMedCrossRefGoogle Scholar
  40. Pietrzykowski AZ, Friesen RM, Martin GE, Puig SI, Nowak CL, Wynne PM, Siegelmann HT, Treistman SN (2008) Posttranscriptional regulation of BK channel spice variant stability by miR-9 underlies neuroadaptation to alcohol. Neuron 59:274–287PubMedCentralPubMedCrossRefGoogle Scholar
  41. Riaz A, Faingold CL (1994) Seizures during ethanol withdrawal are blocked by focal microinjection of excitant amino acid antagonists in the inferior colliculus and pontine reticular formation. Alcohol Clin Exp Res 18:1456–1462PubMedCrossRefGoogle Scholar
  42. Sah P (1996) Ca2+-activated K+ currents in neurones: types, physiological roles and modulation. Trends Neurosci 19:150–154PubMedCrossRefGoogle Scholar
  43. Sheehan JJ, Benedetti BL, Barth AL (2009) Anticonvulsant effects of the BL-channel antagonist paxilline. Epilepsia 50:711–720Google Scholar
  44. Shruti S, Clem LR, Barth AL (2008) A seizure-induced gain-of-function in BK channels is associated with elevated firing activity in neocortical pyramidal neurons. Neurobiol Dis 30:323–330PubMedCentralPubMedCrossRefGoogle Scholar
  45. Shruti S, Urban-Ciecko, Fitzpatrick JA, Brenner R, Bruchez MP, Barth AL (2012) The brain-specific beta4 subunit downregulates BK channel cell surface expression. PLoS ONE 7:e33429. doi: 10.1371/journal.pone.0033429 PubMedCentralPubMedCrossRefGoogle Scholar
  46. Sivaramakrishnan S, Oliver DL (2001) Distinct K+ currents result in physiological distinct cell types in inferior colliculus of the rat. J Neurosci 21:2861–2877PubMedGoogle Scholar
  47. Song X, Su W, Chen L, Ji JJ (2010) Functional expression of large-conductance Ca2+-activated potassium channels in lateral globus pallidus neurons. Neuroscience 169:1548–1556PubMedCrossRefGoogle Scholar
  48. Stern MD (1992) Buffering of calcium in the vicinity of channel pore. Cell Calcium 13:183–192PubMedCrossRefGoogle Scholar
  49. Storm JF (1987) Action potential repolarization and a fst afterhyperpolarization in rat hippocampal pyramidal cells. J Physiol Lond 385:733–759PubMedCentralPubMedGoogle Scholar
  50. Su W, Song X, Ji JJ (2010) Functional expression of a large-conductance Ca2+-activated K+ channel in mouse substantia nigra pars compacta dopaminergic neurons. Neurosci Lett 471:1–5PubMedCrossRefGoogle Scholar
  51. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of protein form polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4354PubMedCentralPubMedCrossRefGoogle Scholar
  52. Veraga C, Latorre R, Marrion NV, Andelman JP (1998) Calcium-activated potassium channels. Curr Opin Neurobiol 8:321–329CrossRefGoogle Scholar
  53. Ye H, Buttigieg J, Wan Y, Wang J, Figley S, Fehlings MG (2012) Expression and functional role of BK channels in chronically injured spinal cord white matter. Neurobiol Dis 47:225–236PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.Department of PediatricsGeorgetown University Medical CenterWashingtonUSA
  2. 2.Department of PharmacologyGeorgetown University Medical CenterWashingtonUSA
  3. 3.Cardiac Signaling CenterMedical University of South CarolinaCharlestonUSA

Personalised recommendations