Association Between Adenosine A2A Receptors and Connexin 43 Regulates Hemichannels Activity and ATP Release in Astrocytes Exposed to Amyloid-β Peptides

Abstract

Increasing evidence implicates astrocytes and the associated purinergic modulation in Alzheimer's disease (AD), characterized by cognitive deficits involving the extracellular amyloid-β peptides (Aβ) accumulation. Aβ can affect astrocytic gliotransmitters release, namely ATP, which is rapidly metabolized into adenosine by ecto-5'-nucleotidase, CD73, resulting in adenosine A_2A receptors (A_2AR) activation that bolsters neurodegeneration. AD's brains exhibit an upregulation of A_2AR and of connexin 43 (Cx43), which in astrocytes forms hemichannels that can mediate ATP release. However, a coupling between astrocytic A_2AR and Cx43 remains to be established. This was now investigated using astrocytic primary cultures exposed to Aβ_1-42 peptides. Aβ triggered ATP release through Cx43 hemichannels, a process blocked by A_2AR antagonists and mimicked by selective A_2AR activation. A_2AR directly regulated hemichannels activity and prevented Cx43 upregulation and phosphorylation observed in Aβ_1-42-exposed astrocytes. Moreover, a proximity ligand assay revealed a physical association between astrocytic A_2AR and Cx43. Finally, the blockade of CD73-mediated extracellular formation of ATP-derived adenosine prevented the Aβ-induced increase of Cx43 hemichannel activity and of ATP release. Overall, the data identify a feed-forward loop involving astrocytic A_2AR and Cx43 hemichannels, whereby A_2AR increase Cx43 hemichannel activity leading to increased ATP release, which is converted into adenosine by CD73, sustaining the increased astrocytic A_2AR activity in AD-like conditions.

Keywords: ATP; Adenosine A2A receptors; Amyloid-β; Astrocytes; CD73; Connexin 43; Hemichannels.