Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 354, Issue 4, pp 420–423 | Cite as

Effect of glutamate receptor ligands on mitochondrial membrane potential in rat dissociated cerebellar cells

  • Francesc X. Sureda
  • Elena Escubedo
  • Cecília Gabriel
  • Jorge Camarasa
  • Antonio Camins
Original Article


The effect of three different glutamate receptor ligands on mitochondrial membrane potential has been studied in rat pup dissociated cerebellar cells by measuring rhodamine 123 fluorescence. L-glutamate, NMDA (N-methyl-D-aspartate) and kainate (from 10−8 to 10−3 M) decreased in a concentration-dependent manner the mitochondrial membrane potential with EC50 values of 6.7±1.7, 3.8±0.5, and 37.4±14 μM, respectively.

Dizocilpine ((+)MK 801) was able to inhibit the NMDA- and L-glutamate-induced decrease in rhodamine 123 fluorescence, while kainate-induced fluorescence-decreases were unaffected. However, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) totally prevented the effect of kainate on mitochondrial membrane potential, but failed to block the L-glutamate effect. It is concluded that, in our cell preparation, L-glutamate exerts its action mainly through NMDA-subtype receptors, and that Ca2+ and Na+ entry through ionotropic glutamate receptors could be responsible for an impairment of mitochondrial membrane potential.

Key words

N-methyl-D-aspartate Glutamate Kainate Dissociated cerebellar cells Rat Mitochondrial membrane potential 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ankarcrona M, Dypbukt JM, Bonfoco E, Zhivotovsky B, Orrenius S, Lipton SA, Nicotera P (1995) Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function. Neuron 15:961–973Google Scholar
  2. Auer RN (1995) Combination therapy with U74006F (tirilazad mesylate), MK-801, insulin and diazepam in transient forebrain ischaemia. Neurol Res 17:132–136Google Scholar
  3. Bowling AC, Beal MF (1995) Bioenergetic and oxidative stress in neurodegenerative diseases. Life Sci 56:1151–1171Google Scholar
  4. Chen LB (1988) Mitochondrial membrane potential in living cells. Ann Rev Cell Biol 4:155–181Google Scholar
  5. Choi DW (1995) Calcium: still center-stage in hypoxic-ischemic neuronal death. Trends Neurosci 18:58–60Google Scholar
  6. Choi DW (1992) Excitotoxic cell death. J Neurobiol 23:1261–1276Google Scholar
  7. Coyle JT, Puttfarcken P (1993) Oxidative stress, glutamate, and neurodegenerative disorders. Science 262:689–695Google Scholar
  8. Di Porzio U, Smith SV, Novotny EA, Morelli F, Barker JL (1993) Two functionally different glutamate receptors of the kainate subtype in embryonic rat mesencephalic cells. Exp Neurol 120:202–213Google Scholar
  9. Domenici MR, Longo R, Scotti de Carolis A, Frank C, Sagratella S (1993) Protective actions of 21-aminosteroids and MK-801 on hypoxia-induced electrophysiological changes in rat hippocampal slices. Eur J Pharmacol 233:291–293Google Scholar
  10. Dubinsky JM (1993) Examination of the role of calcium in neuronal death. Ann NY Acad Sci 679:34–42Google Scholar
  11. Dykens JA (1994) Isolated cerebral and cerebellar mitochondria produce free radicals when exposed to elevated Ca2+ and Na2+: implications for neurodegeneration. J Neurochem 63:584–591Google Scholar
  12. Frandsen A, Drejer J, Schousboe A (1989) Direct evidence that excitotoxicity in cultured neurons is mediated via N-methyl-D-aspartate (NMDA) as well as non-NMDA receptors. J Neurochem 53:297–299Google Scholar
  13. Grierson JP, Petroski RE, O'Connell SM, Geller HM (1992) Calcium homeostasis in dissociated embryonic neurons: a flow cytometric analysis. J Neurophysiol 67:704–714Google Scholar
  14. Hirose K, Chan PH (1993) Blockade of glutamate excitotoxicity and its clinical applications. Neurochem Res 18:479–483Google Scholar
  15. Holopainen I, Enkvist MOK, Akerman KEO (1989) Glutamate receptor agonists increase intracellular Ca2+ independently of voltage-gated Ca2+ channels in rat cerebellar granule cells. Neurosci Lett 98:57–62Google Scholar
  16. Jesberger JA, Richardson JS (1991) Oxygen free radicals and brain dysfunction. Int J Neurosci 57:1–17Google Scholar
  17. Lopez Lozano JJ, Notter MFD, Gash DM, Leary JF (1989) Selective flow cytometric sorting of viable dopamine neurons. Brain Res 486:351–356Google Scholar
  18. Mandler RN, Schaffner AE, Novotny EA, Lange GD, Barker JL (1988) Flow cytometric analysis of membrane potential in embryonic rat spinal cord cells. J Neurosci Methods 22:203–213Google Scholar
  19. Marcaida G, Miana MD, Grisolfa S, Felipo V (1995) Lack of correlation between glutamate-induced deplection of ATP and neuronal death in primary cultures of cerebellum. Brain Res 695:146–150Google Scholar
  20. Monyer H, Hartley DM, Choi DW (1990) 21-Aminosteroids attenuate excitotoxic neuronal injury in cortical cell cultures. Neuron 5:121–126Google Scholar
  21. Nicotera P, Bellomo G, Orrenius S (1992) Calcium-mediated mechanisms in chemically induced cell death. Annu Rev Pharmacol Toxicol 32:449–470Google Scholar
  22. Oyama Y, Hayashi A, Ueha T, Maekawa K (1995) Characterization of 2′,7′-dichlorofluorescin fluorescence in dissociated mammalian brain neurons: estimation on intracellular content of hydrogen peroxide. Brain Res 635:113–117Google Scholar
  23. Rogawski MA (1995) The NMDA receptor, NMDA antagonists and epilepsy therapy. Drugs 44:279–292Google Scholar
  24. Schieren I, MacDermott A (1988) Flow cytometric identification and purification of cells by ligand-induced changes in intracellular calcium. J Neurosci Methods 26:35–44Google Scholar
  25. St. John PA, Kell WM, Mazzetta JS, Lange GD, Barker JL (1986) Analysis and isolation of embryonic mammalian neurons by fluorescence activated cell sorting. J Neurosci 6:1492–1512Google Scholar
  26. Sureda FX, Camins A, Trullas R, Camarasa J, Escubedo E (1996) A flow cytometric study of N-methyl-D-aspartate effects on dissociated cerebellar cells. Brain Res (in press)Google Scholar
  27. Taylor CP, Meldrum BS (1995) Na+ channels as targets for neuroprotective drugs. Trends Pharmacol Sci 16:309–316Google Scholar
  28. Trump BF, Berezesky IK (1995) Calcium-mediated cell injury and cell death. FASEB J 9:219–228Google Scholar
  29. Wong EHF, Kemp J (1991) Sites for antagonism on the N-methyl-D-aspartate receptor channel complex. Annu Rev Pharmacol Toxicol 31:401–425Google Scholar
  30. Wolf ME, Kapatos G (1989) Flow cytometric analysis of rat striatal nerve terminals. J Neurosci 9:94–105Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Francesc X. Sureda
    • 1
  • Elena Escubedo
    • 1
  • Cecília Gabriel
    • 1
  • Jorge Camarasa
    • 1
  • Antonio Camins
    • 1
  1. 1.Unitat de Farmacologia i Farmacognòsia, Facultat de FarmàciaUniversitat de BarcelonaBarcelonaSpain

Personalised recommendations