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Neurochemical Research

, Volume 43, Issue 1, pp 79–88 | Cite as

Alleviation by GABAB Receptors of Neurotoxicity Mediated by Mitochondrial Permeability Transition Pore in Cultured Murine Cortical Neurons Exposed to N-Methyl-d-aspartate

  • Toshihiko Kinjo
  • Yoshino Ashida
  • Hiroshi Higashi
  • Satoshi Sugimura
  • Miho Washida
  • Hiroki Niihara
  • Kiyokazu Ogita
  • Yukio Yoneda
  • Nobuyuki KuramotoEmail author
Original Paper
  • 189 Downloads

Abstract

Mitochondrial permeability transition pore (PTP) is supposed to at least in part participate in molecular mechanisms underlying the neurotoxicity seen after overactivation of N-methyl-d-aspartate (NMDA) receptor (NMDAR) in neurons. In this study, we have evaluated whether activation of GABAB receptor (GABABR), which is linked to membrane G protein-coupled inwardly-rectifying K+ ion channels (GIRKs), leads to protection of the NMDA-induced neurotoxicity in a manner relevant to mitochondrial membrane depolarization in cultured embryonic mouse cortical neurons. The cationic fluorescent dye 3,3′-dipropylthiacarbocyanine was used for determination of mitochondrial membrane potential. The PTP opener salicylic acid induced a fluorescence increase with a vitality decrease in a manner sensitive to the PTP inhibitor ciclosporin, while ciclosporin alone was effective in significantly preventing both fluorescence increase and viability decrease by NMDA as seen with an NMDAR antagonist. The NMDA-induced fluorescence increase and viability decrease were similarly prevented by pretreatment with the GABABR agonist baclofen, but not by the GABAAR agonist muscimol, in a fashion sensitive to a GABABR antagonist. Moreover, the GIRK inhibitor tertiapin canceled the inhibition by baclofen of the NMDA-induced fluorescence increase. These results suggest that GABABR rather than GABAAR is protective against the NMDA-induced neurotoxicity mediated by mitochondrial PTP through a mechanism relevant to opening of membrane GIRKs in neurons.

Keywords

GABABGIRKs NMDAR Mitochondrial depolarization Cell death Tertiapin 

Abbreviations

AC

Adenylyl cyclase

AUC

Area under curve

DiSC3(5)

3,3′-Dipropylthiacarbocyanine iodide

DIV

Days in vitro

GABA

γ-Aminobutyric acid

GABAAR

γ-Aminobutyric acid A receptor

GABABR

γ-Aminobutyric acid B receptor

GIRK

G protein-coupled inwardly-rectifying K+ ion channel

GPCR

G protein-coupled receptor

MTT

3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide

NMDA

N-Methyl-d-aspartate

NMDAR

N-Methyl-d-aspartate receptor

MK-801

Dizocilpine

PKA

Protein kinase A

PTP

Permeability transition pore

ROS

Reactive oxygen species

SQ22536

9-(Tetrahydro-2-furanyl)-9H-purin-6-amine, 9-THF-Ade

Notes

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Fujikawa DG (2005) Prolonged seizures and cellular injury: understanding the connection. Epilepsy Behav 7:S3–S11CrossRefGoogle Scholar
  2. 2.
    Halenbeck R, MacDonald H, Roulston A, Chen TT, Conroy L, Williams LT (1998) CPAN, a human nuclease regulated by the caspase-sensitive inhibitor DFF45. Curr Biol 8:537–540CrossRefGoogle Scholar
  3. 3.
    Parrish J, Li L, Klotz K, Ledwich D, Wang X, Xue D (2001) Mitochondrial endonuclease G is important for apoptosis in C. elegans. Nature 412:90–94CrossRefGoogle Scholar
  4. 4.
    Halestrap AP, Doran E, Gillespie JP, O’Toole A (2000) Mitochondria and cell death. Biochem Soc Trans 28:170–177CrossRefGoogle Scholar
  5. 5.
    Bernardi P (1992) Modulation of the mitochondrial cyclosporin A-sensitive permeability transition pore by the proton electrochemical gradient. Evidence that the pore can be opened by membrane depolarization. J Biol Chem 267:8834–8839PubMedGoogle Scholar
  6. 6.
    Bernardi P, Broekemeier KM, Pfeiffer DR (1994) Recent progress on regulation of the mitochondrial permeability transition pore; a cyclosporin-sensitive pore in the inner mitochondrial membrane. J Bioenerg Biomembr 26(5):509–517CrossRefGoogle Scholar
  7. 7.
    Bettler B, Kaupmann K, Mosbacher J, Gassmann M (2004) Molecular structure and physiological functions of GABA(B) receptors. Physiol Rev 84:835–867CrossRefGoogle Scholar
  8. 8.
    Deng PY, Xiao Z, Yang C, Rojanathammanee L, Grisanti L, Watt J, Geiger JD, Liu R, Porter JE, Lei S (2009) GABA(B) receptor activation inhibits neuronal excitability and spatial learning in the entorhinal cortex by activating TREK-2 K+ channels. Neuron 63(2):230–243CrossRefGoogle Scholar
  9. 9.
    Chalifoux JR, Carter AG (2010) GABAB receptors modulate NMDA receptor calcium signals in dendritic spines. Neuron 66:101–113CrossRefGoogle Scholar
  10. 10.
    Kazmierska P, Konopacki J (2013) Development of NMDA-induced theta rhythm in hippocampal formation slices. Brain Res Bull 98:93–101CrossRefGoogle Scholar
  11. 11.
    di Porzio U, Daguet MC, Glowinski J, Prochiantz A (1980) Effect of striatal cells on in vitro maturation of mesencephalic dopaminergic neurones grown in serum-free conditions. Nature 288:370–373CrossRefGoogle Scholar
  12. 12.
    Plásek J, Sigler K (1996) Slow fluorescent indicators of membrane potential: a survey of different approaches to probe response analysis. J Photochem Photobiol B 33:101–124CrossRefGoogle Scholar
  13. 13.
    Klein B, Worndl K, Lutz-Meindl U, Kerschbaum HH (2011) Perturbation of intracellular K+ homeostasis with valinomycin promotes cell death by mitochondrial swelling and autophagic processes. Apoptosis 16:1101–1117CrossRefGoogle Scholar
  14. 14.
    Hrynevich SV, Pekun TG, Waseem TV, Fedorovich SV (2015) Influence of glucose deprivation on membrane potentials of plasma membranes, mitochondria and synaptic vesicles in rat brain synaptosomes. Neurochem Res 40:1188–1196CrossRefGoogle Scholar
  15. 15.
    Salvioli S, Ardizzoni A, Franceschi C, Cossarizza A (1997) JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: implications for studies on mitochondrial functionality during apoptosis. FEBS Lett 411:77–82CrossRefGoogle Scholar
  16. 16.
    A Szewczyka, J Skalskaa, M Głąba, B Kulawiaka, D Malińskaa, I Koszela-Piotrowskaa, WS. Kunzb (2006) Mitochondrial potassium channels: from pharmacology to function. Biochim Biophys Acta 1757:715–720CrossRefGoogle Scholar
  17. 17.
    Szydlowska K, Tymianski M (2010) Calcium, ischemia and excitotoxicity. Cell Calcium 47:122–129CrossRefGoogle Scholar
  18. 18.
    Pinton P, Giorgi C, Siviero R, Zecchini E, Rizzuto R (2008) Calcium and apoptosis: ER-mitochondria Ca2+ transfer in the control of apoptosis. Oncogene 27:6407–6418CrossRefGoogle Scholar
  19. 19.
    Schinder AF, Olson EC, Spitzer NC, Montal M (1996) Mitochondrial dysfunction is a primary event in glutamate neurotoxicity. J Neurosci 16:6125–6133CrossRefGoogle Scholar
  20. 20.
    Raiteri M (2006) Functional pharmacology in human brain. Pharmacol Rev 58:162–193CrossRefGoogle Scholar
  21. 21.
    Federici M, Nisticò R, Giustizieri M, Bernardi G, Mercuri NB (2009) Ethanol enhances GABAB-mediated inhibitory postsynaptic transmission on rat midbrain dopaminergic neurons by facilitating GIRK currents. Eur J Neurosci 29:1369–1377CrossRefGoogle Scholar
  22. 22.
    Laskowski M, Augustynek B, Kulawiak B, Koprowski P, Bednarczyk P, Jarmuszkiewicz W, Szewczyk A. (2016) What do we not know about mitochondrial potassium channels? Biochim Biophys Acta. 1857:1247–1257CrossRefGoogle Scholar
  23. 23.
    Jin W, Lu Z (1998) A novel high-affinity inhibitor for inward-rectifier K+ channels. BioChemistry 37:13291–13299CrossRefGoogle Scholar
  24. 24.
    Kanjhan R, Coulson EJ, Adams DJ, Bellingham MC (2005) Tertiapin-Q blocks recombinant and native large conductance K+ channels in a use-dependent manner. J Pharmacol Exp Ther 314:1353–1361CrossRefGoogle Scholar
  25. 25.
    Yamada K, Inagaki N (2005) Neuroprotection by KATP channels. J Mol Cell Cardiol 38:945–949CrossRefGoogle Scholar
  26. 26.
    Heurteaux C, Bertaina V, Widmann C, Lazdunski M (1993) K+ channel openers prevent global ischemia-induced expression of c-fos, c-jun, heat shock protein, and amyloid beta-protein precursor genes and neuronal death in rat hippocampus. Proc Natl Acad Sci USA 90:9431–9435CrossRefGoogle Scholar
  27. 27.
    Kong J, Ren G, Jia N, Wang Y, Zhang H, Zhang W, Chen B, Cao Y (2013) Effects of nicorandil in neuroprotective activation of PI3K/AKT pathways in a cellular model of Alzheimer’s disease. Eur Neurol 70:233–241CrossRefGoogle Scholar
  28. 28.
    Lüscher C, Slesinger PA (2010) Emerging roles for G protein-gated inwardly rectifying potassium (GIRK) channels in health and disease. Nat Rev Neurosci 11:301–315CrossRefGoogle Scholar
  29. 29.
    Nagy K, Kis B, Rajapakse NC, Bari F, Busija DW (2004) Diazoxide preconditioning protects against neuronal cell death by attenuation of oxidative stress upon glutamate stimulation. J Neurosci Res 76:697–704CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Laboratory of Molecular Pharmacology, Faculty of Pharmaceutical SciencesSetsunan UniversityHirakataJapan
  2. 2.Laboratory of Pharmacology, Faculty of Pharmaceutical SciencesSetsunan UniversityHirakataJapan
  3. 3.Section of Prophylactic Pharmacology, Venture Business LaboratoryKanazawa UniversityKanazawaJapan

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