Keywords :
Ion Channels; Adenosine Triphosphate; Potassium; Calcium; Cells, Cultured; Electrophysiology; Immunohistochemistry; Neuroglia/drug effects; Neuroglia/metabolism; Neuromuscular Junction/drug effects; Patch-Clamp Techniques; ATP; Ca2+-activated K+ current; Cl- conductance; P2X7 receptor; Schwann cell; Neuroscience (all); General Neuroscience
Abstract :
[en] Extracellular ATP, by acting on P2 purinergic receptors, is a potent mediator of cell-to-cell communication both within and between the nervous and the immune systems. We show here by patch-clamp recording, fluorescent dye uptake and immunocytochemistry that, in cultured mouse Schwann cells, ATP activates a P2X(7) receptor associated with three different ionic conductances. In control conditions, ATP activated an inward current (I(ATP)) with a low potency (EC(50), 7.2 mM). Replacing ATP either by the ATP analogue 2',3'-O-(4-benzoyl-4-benzoyl)-ATP (BzATP) or by the tetraacidic form ATP(4-) potentiated the inward current (ATP(4-) EC(50), 375 microM). ATP and BzATP currents were strongly reduced by periodate oxidized ATP (oATP), an antagonist of P2X(7) receptors. IATP was a mixed current composed of a nonselective cationic conductance, a cationic conductance selective for K(+) and an anionic conductance selective for Cl(-). The activation of the K(+) conductance was dependent on an influx of Ca(2+), and was blocked by charybdotoxin (ChTX) and tetraethylammonium (TEA), two potent antagonists of large conductance Ca(2+)-activated K(+) channels (BK channels). The activation of the Cl(-) conductance was insensitive to Ca(2+) but required the presence of K(+). Total removal of K(+) blocked both the Ca(2+)-activated K(+) conductance and the Cl(-) conductance, unveiling the P2X(7) nonselective cationic conductance. The P2X(7) receptor was localized by immunocytochemistry using a polyclonal antibody, anti-P2X(7), whilst its expression and functionality were both detected by the uptake of Lucifer Yellow. This receptor could regulate the synthesis and the release of cytokines by Schwann cells during pathophysiological events.
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