In the brain, endfeet of perivascular astrocytes make close contact with capillary endothelial cells that form the blood-brain barrier. The aim of the present work was to investigate whether and how calcium signals can be communicated from astrocytes to endothelial cells following acute mechanical cell damage. The experiments were performed on astrocyte-endothelial co-cultures prepared from primary rat brain astrocytes and an endothelial cell line (ECV304). A single astrocyte was acutely damaged by mechanical stimulation of sufficient strength with a micropipette, and the resulting cytoplasmic calcium changes were monitored using fura-2 and digital calcium imaging. Mechanical damage to a single astrocyte triggered a large intercellular calcium wave that propagated to surrounding astrocytes and also to even remotely located (several hundred micrometers) endothelial cells. Astrocyte-endothelial calcium waves induced by mechanical cell damage were largely deflected by fast superfusion, were able to cross a cell-free lane, were dose-dependently inhibited by suramin, a P2-purinoceptor blocker, and were largely reduced in size in the presence of the ATP-degrading enzyme apyrase. Our results indicate that mechanical damage to a single astrocyte can produce far reaching calcium signals that are propagated by the release of a calcium mobilizing P2-purinergic agonist and that can be communicated to endothelial cells. As endothelial cytoplasmic calcium is an important factor in the regulation of blood-brain barrier permeability and transport, mechanical cell damage-induced astrocyte-endothelial calcium signals are hypothesized to play a role in the initiation of brain edema and the stimulation of brain glucose uptake.