Allosteric modulation of GABA_A receptors by extracellular ATP

The γ-aminobutyric acid type A receptor (GABA_AR) is the primary receptor mediating fast synaptic inhibition in the brain and plays a critical role in modulation of neuronal excitability and neural networks. Previous studies have demonstrated that ATP and its nucleotide analogs may regulate the function of GABA_ARs via Ca²⁺-dependent intracellular mechanisms, which require activation of purinergic 2 (P2) receptors or cross-talk between two receptors.

Here, we report a potentiation of GABA_ARs by extracellular ATP via a previously un-recognized allosteric mechanism. Using cultured hippocampal neurons as well as HEK293 cells transiently expressing GABA_ARs, we demonstrate that extracellular ATP potentiates GABA_AR mediated currents in a dose-dependent manner with an EC50 of 2.1 ± 0.2 mM. The potentiation was mediated by a postsynaptic mechanism that was not dependent on activation of either ecto-protein kinase or P2 receptors. Single channel recordings from cell-free excised membrane patches under outside-out mode or isolated membrane patches under cell-attached mode suggest that the ATP modulation of GABA currents is achieved through a direct action of ATP on the channels themselves and manifested by increasing the single channel open probability without alteration of its conductance. Moreover, this ATP potentiation of GABA_AR could be reconstituted in HEK293 cells that transiently expressed recombinant rat GABA_ARs.

Our data strongly suggest that extracellular ATP allosterically potentiates GABA_AR-gated chloride channels. This novel mode of ATP-mediated modulation of GABA_ARs may play an important role in regulating neuronal excitability and thereby in fine-tuning the excitation-inhibition balance under conditions where a high level of extracellular ATP is ensured.