Expression and functional characterization of P2X receptors in mouse alveolar macrophages

Abstract

Alveolar macrophages (AM) are crucial for pulmonary host defense, and evidence emerges that ATP-gated P2X receptors are involved in inflammatory processes. This study focuses on the expression and functional characterization of P2X receptors in AM from mouse. In RT-PCR experiments, transcripts encoding the P2X₁, P2X₃, P2X₄, P2X₅, and P2X₇ receptors were detected. In whole-cell patch-clamp recordings, ATP (1 mM) evoked an inward current (mouse and human AM) that was reversible upon washout, and the reversal potential was ~5 mV, indicating the activation of a non-selective conductance-a fingerprint of P2X receptors. Further characterization (mouse AM) revealed that the current was not desensitized by a second ATP application. The ATP-induced current was increased by the removal of extracellular Ca²⁺ (in human and mouse AM), and EC₅₀ in mouse AM were determined with ~1 mM ATP, in the presence as well as in the absence of extracellular Ca²⁺. Pharmacological characterization of mouse AM revealed that the effect was augmented by BzATP and pre-application with ivermectin, but no effect with α,β-meATP was observed. Further, the ATP effect was reduced by PPADS (300 μM), brilliant blue G (5 μM), and about A438079 (10 μM). Although different P2X receptor transcripts were detected in mouse AM, the observed functional and pharmacological characteristics indicate primarily the participation of P2X₄ and P2X₇ receptors as mediators of the ATP-induced ion current in mouse AM. These suggestions were confirmed by experiments with AM from P2X₇ -deficient animals, indicating a contribution of P2X₄ and P2X₇ receptors in pulmonary immune function.