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Regulation of intracellular Ca 2+ by P2Y 1 receptors may depend on the developmental stage of cultured rat striatal neurons
Author(s) -
Rubini Patrizia,
Pinkwart Christina,
Franke Heike,
Gerevich Zoltan,
Nörenberg Wolfgang,
Illes Peter
Publication year - 2006
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.20705
Subject(s) - ppads , depolarization , p2y receptor , intracellular , cyclopiazonic acid , patch clamp , receptor , biophysics , nmda receptor , membrane potential , biology , fura 2 , muscimol , purinergic receptor , calcium in biology , chemistry , microbiology and biotechnology , biochemistry , agonist , cytosol , enzyme
Mixed striatal cell cultures containing neurons and glial cells were grown either in neurobasal medium (NBM) or Dulbecco's modified Eagle's medium (DMEM). Whole‐cell patch‐clamp recordings indicated that, if at all, only a single, low amplitude spike was evoked shortly after starting the injection of a depolarizing current pulse into NBM neurons. In contrast, DMEM neurons fired series of high amplitude action potentials, without apparent spike frequency adaptation. The possible reason for the observed action potential failure in NBM neurons was a low density of Na + channels per unit of membrane surface area. However, both in NBM and DMEM neurons, ATP did not induce inward current responses via P2X receptor‐channels, although GABA A and N ‐methyl‐ D ‐aspartate (NMDA) receptor‐channels could be activated by muscimol and NMDA, respectively. Ca 2+ imaging experiments by means of the Fura‐2 method were utilized to measure intracellular Ca 2+ ([Ca 2+ ] i ) in neurons and glial cells. NBM, but not DMEM neurons responded to ATP with [Ca 2+ ] i transients; glial cells grown in either culture medium were equally sensitive to ATP. ATP caused an increase of [Ca 2+ ] i by a mechanism only partly dependent on external Ca 2+ ; the residual ATP effect was blocked by cyclopiazonic acid (CPA) and was therefore due to the release of Ca 2+ from its intracellular pools. The receptor involved was characterized by P2 receptor antagonists (PPADS, MRS 2179, AR‐C69931MX) and was found to belong to the P2Y 1 subtype. CPA caused an early [Ca 2+ ] i response due to release from intracellular storage sites, followed by a late [Ca 2+ ] i response due to the influx of this cation from the extracellular space, probably triggered by the opening of store‐operated channels (SOCs) in the plasma membrane. It is concluded that in partial analogy with the effect of CPA, ATP releases [Ca 2+ ] i via the G q /phospholipase C/inositoltrisphosphate (IP 3 ) pathway, thereby opening SOCs. It is hypothesized that this effect of ATP may have an important role for the proliferation and migration of striatal neuronal progenitors. J. Cell. Physiol. 209: 81–93, 2006. © 2006 Wiley‐Liss, Inc.