Supersensitivity of P2X7 receptors in cerebrocortical cell cultures after in vitro ischemia

Kerstin Wirkner, Attila Köfalvi, Wolfgang Fischer, Albrecht Günther, Heike Franke, Heike Gröger-Arndt, Wolfgang Nörenberg, E. Madarász, E. Vízi, Dietmar Schneider, B. Sperlágh, Peter Illes

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Neuronally enriched primary cerebrocortical cultures were exposed to glucose-free medium saturated with argon (in vitro ischemia) instead of oxygen (normoxia). Ischemia did not alter P2X7 receptor mRNA, although serum deprivation clearly increased it. Accordingly, P2X7 receptor immunoreactivity (IR) of microtubuline-associated protein 2 (MAP2)-IR neurons or of glial fibrillary acidic protein (GFAP)-IR astrocytes was not affected; serum deprivation augmented the P2X7 receptor IR only in the astrocytic, but not the neuronal cell population. However, ischemia markedly increased the ATP- and 2′-3′-O-(4-benzoylbenzoyl)-adenosine 5′-triphosphate (BzATP)-induced release of previously incorporated [3H]GABA. Both Brilliant Blue G and oxidized ATP inhibited the release of [3H]GABA caused by ATP application; the Brilliant Blue G-sensitive, P2X7 receptor-mediated fraction, was much larger after ischemia than after normoxia. Whereas ischemic stimulation failed to alter the amplitude of ATP- and BzATP-induced small inward currents recorded from a subset of non-pyramidal neurons, BzATP caused a more pronounced increase in the frequency of miniature inhibitory postsynaptic currents (mlPSCs) after ischemia than after normoxia. Brilliant Blue G almost abolished the effect of BzATP in normoxic neurons. Since neither the amplitude of mlPSCs nor that of the muscimol-induced inward currents was affected by BzATP, it is assumed that BzATP acts at presynaptic P2X7 receptors. Finally, P2X7 receptors did not enhance the intracellular free Ca2+ concentration either in proximal dendrites or in astrocytes, irrespective of the normoxic or ischemic pre-incubation conditions. Hence, facilitatory P2X7 receptors may be situated at the axon terminals of GABAergic non-pyramidal neurons. When compared with normoxia, ischemia appears to markedly increase P2X7 receptor-mediated GABA release, which may limit the severity of the ischemic damage. At the same time we did not find an accompanying enhancement of P2X 7 mRNA or protein expression, suggesting that receptors may become hypersensitive because of an increased efficiency of their transduction pathways.

Original languageEnglish
Pages (from-to)1421-1437
Number of pages17
JournalJournal of Neurochemistry
Volume95
Issue number5
DOIs
Publication statusPublished - Dec 2005

Fingerprint

Purinergic P2X7 Receptors
Cell culture
Ischemia
Cell Culture Techniques
Adenosine
Adenosine Triphosphate
Neurons
gamma-Aminobutyric Acid
Astrocytes
Presynaptic Receptors
Inhibitory Postsynaptic Potentials
Muscimol
Messenger RNA
In Vitro Techniques
Argon
Glial Fibrillary Acidic Protein
Induced currents
Presynaptic Terminals
Dendrites
Serum

Keywords

  • ATP
  • Cortical cell culture
  • Ischemia
  • P2X receptor
  • Receptor up-regulation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Wirkner, K., Köfalvi, A., Fischer, W., Günther, A., Franke, H., Gröger-Arndt, H., ... Illes, P. (2005). Supersensitivity of P2X7 receptors in cerebrocortical cell cultures after in vitro ischemia. Journal of Neurochemistry, 95(5), 1421-1437. https://doi.org/10.1111/j.1471-4159.2005.03465.x

Supersensitivity of P2X7 receptors in cerebrocortical cell cultures after in vitro ischemia. / Wirkner, Kerstin; Köfalvi, Attila; Fischer, Wolfgang; Günther, Albrecht; Franke, Heike; Gröger-Arndt, Heike; Nörenberg, Wolfgang; Madarász, E.; Vízi, E.; Schneider, Dietmar; Sperlágh, B.; Illes, Peter.

In: Journal of Neurochemistry, Vol. 95, No. 5, 12.2005, p. 1421-1437.

Research output: Contribution to journalArticle

Wirkner, K, Köfalvi, A, Fischer, W, Günther, A, Franke, H, Gröger-Arndt, H, Nörenberg, W, Madarász, E, Vízi, E, Schneider, D, Sperlágh, B & Illes, P 2005, 'Supersensitivity of P2X7 receptors in cerebrocortical cell cultures after in vitro ischemia', Journal of Neurochemistry, vol. 95, no. 5, pp. 1421-1437. https://doi.org/10.1111/j.1471-4159.2005.03465.x
Wirkner, Kerstin ; Köfalvi, Attila ; Fischer, Wolfgang ; Günther, Albrecht ; Franke, Heike ; Gröger-Arndt, Heike ; Nörenberg, Wolfgang ; Madarász, E. ; Vízi, E. ; Schneider, Dietmar ; Sperlágh, B. ; Illes, Peter. / Supersensitivity of P2X7 receptors in cerebrocortical cell cultures after in vitro ischemia. In: Journal of Neurochemistry. 2005 ; Vol. 95, No. 5. pp. 1421-1437.
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