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Supersensitivity of P2X 7 receptors in cerebrocortical cell cultures after in vitro ischemia
Author(s) -
Wirkner Kerstin,
Köfalvi Attila,
Fischer Wolfgang,
Günther Albrecht,
Franke Heike,
GrögerArndt Helke,
Nörenberg Wolfgang,
Madarász Emília,
Vizi E. Sylvester,
Schneider Dietmar,
Sperlágh Beáta,
Illes Peter
Publication year - 2005
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2005.03465.x
Subject(s) - receptor , glial fibrillary acidic protein , ischemia , astrocyte , stimulation , medicine , biology , biophysics , chemistry , endocrinology , biochemistry , central nervous system , immunohistochemistry
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 P2X 7 receptor mRNA, although serum deprivation clearly increased it. Accordingly, P2X 7 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 P2X 7 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 [ 3 H]GABA. Both Brilliant Blue G and oxidized ATP inhibited the release of [ 3 H]GABA caused by ATP application; the Brilliant Blue G‐sensitive, P2X 7 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 (mIPSCs) after ischemia than after normoxia. Brilliant Blue G almost abolished the effect of BzATP in normoxic neurons. Since neither the amplitude of mIPSCs nor that of the muscimol‐induced inward currents was affected by BzATP, it is assumed that BzATP acts at presynaptic P2X 7 receptors. Finally, P2X 7 receptors did not enhance the intracellular free Ca 2+ concentration either in proximal dendrites or in astrocytes, irrespective of the normoxic or ischemic pre‐incubation conditions. Hence, facilitatory P2X 7 receptors may be situated at the axon terminals of GABAergic non‐pyramidal neurons. When compared with normoxia, ischemia appears to markedly increase P2X 7 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.