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  • Title: P2U nucleotide receptor activation in rat glial cell line induces [Ca2+]i oscillations which depend on cytosolic pH.
    Author: Czubayko U, Reiser G.
    Journal: Glia; 1996 Feb; 16(2):108-16. PubMed ID: 8929898.
    Abstract:
    In single rat glioma cells, the signal transduction process activated by the UTP sensitive purinergic nucleotide receptor was studied by determining [Ca2+]i by Fura-2 fluorescence and measuring pH by BCECF fluorescence to elucidate the control of [Ca2+]i oscillations by intracellular pH. Addition of UTP for long time periods (some min) causes a [Ca2+]i response composed of i) an initial large peak and a following sustained increase (160 s duration), and ii) subsequent regular [Ca2+]i oscillations (amplitude 107 nM, frequency 1.5 oscillations per min). The maintenance of the [Ca2+]i oscillations depends on the continued presence of agonist. The oscillations are abolished by reducing extracellular Ca2+ concentration. The interaction of UTP receptors and bradykinin receptors during the [Ca2+]i oscillations was investigated because previous studies have already shown that the peptide causes comparable [Ca2+]i oscillations. During [Ca2+]i oscillations induced by UTP or bradykinin, long-term admission of both hormones (400-500 s) causes a large initial response superimposed on regular [Ca2+]i oscillations. Short pulses (12 s) of the second agonist given in any phase of the oscillations induce large [Ca2+]i peaks. In both cases, the following oscillations are not disturbed. The influence of cytosolic pH was studied by alkalinizing pHi by application of NH4Cl. [Ca2+]i oscillations stop after addition of NH4Cl. Recovery of NH4Cl-induced alkalinization is reduced by furosemide. To the same degree, the interruption of [Ca2+]i oscillations is significantly prolonged in the presence of furosemide. Thus cytosolic alkalinization suppresses hormone-induced [Ca2+]i oscillations in rat glioma cells. The understanding of the molecular mechanism of this interference of pH should provide an important contribution for unravelling the function of cytosolic pH in cellular signal transduction.
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