Extracellular ATP‐induced currents in astrocytes: Involvement of a cation channel

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Abstract
Whole‐cell currents were measured with the perforated patch clamp technique in cultured rat astrocytes to analyze the underlying ionic mechanism for a P2‐purinoceptor‐mediated depolarization. ATP (100 μM) induced an inward current with a mean amplitude of 130 pA and an EC50 of 17 μM. The response desensitized during a 1 min application. Replacement of extracellular Na+ with NMDG or K+ abolished the ATP‐evoked inward current. Replacement of Na+ with choline, however, resulted in an ATP‐evoked response of one‐third the amplitude in normal solution. This is indicative of a cation rather than Na+ channel. However, due to difficulties in voltage‐clamping these gap junction‐coupled cells at voltages different from the membrane resting potential, the current reversal potential could not be determined. Measurements with K+‐sensitive microelectrodes showed that 100 μM ATP lowered the intracellular K+ concentration. Replacement of extracellular Ca2+ or Cl− did not alter the ATP‐induced inward currents. Fura‐2 imaging experiments revealed a transient rise of the intracellular Ca2+ concentration during ATP application. Removal of extracellular Ca2+ did not influence the peak response; it did, however, shorten the time course. These results and previous observations that the permeability changes are caused by a P2× receptor are indicative of an ATP‐sensitive cation conductance. In addition, cytoplasmic Ca2+ is increased by mobilization from intracellular stores, and by additional influx across the cell membrane. Extracellular ATP released by neurons could evoke K+ release from astrocytes as well as be a mediator for cation changes that signal cell activation processes when released by damaged cells.