Purine receptor mediated actin cytoskeleton remodeling of human fibroblasts

Earlier studies have shown that activation of adenosine A1 receptors on peripheral pain fibers contributes to acupuncture-induced suppression of painful input. In addition to adenosine, acupuncture triggers the release of other purines, including ATP and ADP that may bind to purine receptors on nearby fibroblasts. We here show that purine agonists trigger increase in cytosolic Ca(2+) signaling in a cultured human fibroblasts cell line. The profile of agonist-induced Ca(2+) increases indicates that the cells express functional P2yR2 and P2yR4 receptors, as well as P2yR1 and P2xR7 receptors. Unexpectedly, purine-induced Ca(2+) signaling was associated with a remodeling of the actin cytoskeleton. ATP induced a transient loss in F-actin stress fiber. The changes of actin cytoskeleton occurred slowly and peaked at 10min after agonist exposure. Inhibition of ATP-induced increases in Ca(2+) by cyclopiazonic acid blocked receptor-mediated cytoskeleton remodeling. The Ca(2+) ionophore failed to induce cytoskeletal remodeling despite triggering robust increases in cytosolic Ca(2+). These observations indicate that purine signaling induces transient changes in fibroblast cytoarchitecture that could be related to the beneficial effects of acupuncture.