Role of Purinergic Receptor P2Y1 in Spatiotemporal Ca²⁺Dynamics in Astrocytes

Abstract

Fine processes of astrocytes enwrap synapses and are well positioned to sense neuronal information via synaptic transmission. In rodents, astrocyte processes sense synaptic transmission via Gq-protein coupled receptors (GqPCR), including the P2Y1 receptor (P2Y1R), to generate Ca 2+ signals. Astrocytes display numerous spontaneous microdomain Ca 2+ signals; however, it is not clear whether such signals are due to local synaptic transmission and/or in what timeframe astrocytes sense local synaptic transmission. To ask whether GqPCRs mediate microdomain Ca 2+ signals, we engineered mice (both sexes) to specifically overexpress P2Y1Rs in astrocytes, and we visualized Ca 2+ signals via a genetically encoded Ca 2+ indicator, GCaMP6f, in astrocytes from adult mice. Astrocytes overexpressing P2Y1Rs showed significantly larger Ca 2+ signals in response to exogenously applied ligand and to repetitive electrical stimulation of axons compared with controls. However, we found no evidence of increased microdomain Ca 2+ signals. Instead, Ca 2+ waves appeared and propagated to occupy areas that were up to 80-fold larger than microdomain Ca 2+ signals. These Ca 2+ waves accounted for only 2% of total Ca 2+ events, but they were 1.9-fold larger and 2.9-fold longer in duration than microdomain Ca 2+ signals at processes. Ca 2+ waves did not require action potentials for their generation and occurred in a probenecid-sensitive manner, indicating that the endogenous ligand for P2Y1R is elevated independently of synaptic transmission. Our data suggest that spontaneous microdomain Ca 2+ signals occur independently of P2Y1R activation and that astrocytes may not encode neuronal information in response to synaptic transmission at a point source of neurotransmitter release. SIGNIFICANCE STATEMENT Astrocytes are thought to enwrap synapses with their processes to receive neuronal information via Gq-protein coupled receptors (GqPCRs). Astrocyte processes display numerous microdomain Ca 2+ signals that occur spontaneously. To determine whether GqPCRs play a role in microdomain Ca 2+ signals and the timeframe in which astrocytes sense neuronal information, we engineered mice whose astrocytes specifically overexpress the P2Y1 receptor, a major GqPCR in astrocytes. We found that overexpression of P2Y1 receptors in astrocytes did not increase microdomain Ca 2+ signals in astrocyte processes but caused Ca 2+ wavelike signals. Our data indicate that spontaneous microdomain Ca 2+ signals do not require activation of P2Y1 receptors.