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Wave-like propagation of [Ca 2+ ] i  increases is a remarkable intercellular communication characteristic in astrocyte networks, intercalating neural circuits and vasculature. Mechanically-induced [Ca 2+ ] i  increases and their subsequent propagation to neighboring astrocytes in culture is a classical model of astrocyte calcium wave and is known to be mediated by gap junction and extracellular ATP, but the role of each pathway remains unclear. Pharmacologic analysis of time-dependent distribution of [Ca 2+ ] i  revealed three distinct [Ca 2+ ] i  increases, the largest being in stimulated cells independent of extracellular Ca 2+  and inositol 1,4,5-trisphosphate-induced Ca 2+  release. In addition, persistent [Ca 2+ ] i  increases were found to propagate rapidly via gap junctions in the proximal region, and transient [Ca 2+ ] i  increases were found to propagate slowly via extracellular ATP in the distal region. Simultaneous imaging of astrocyte [Ca 2+ ] i  and extracellular ATP, the latter of which was measured by an ATP sniffing cell, revealed that ATP was released within the proximal region by volume-regulated anion channel in a [Ca 2+ ] i  independent manner. This detailed analysis of a classical model is the first to address the different contributions of two major pathways of calcium waves, gap junctions and extracellular ATP.

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Astrocyte calcium waves propagate proximally by gap junction and distally by extracellular diffusion of ATP released from volume-regulated anion channels