Anales RANF

P.84 INTERACTION OF THE SPHINGOSINE-1-PHOSPHATE PATHWAY WITH PURINERGIC RECEPTORS D. Zahiri 1 , M. Klapperstück 1 , F. Markwardt 1 1 . University of Halle (Saale), Saxony-Anhalt, Germany. Abstract: Cells release ATP in many ways. One of them is via the cell volume-sensitive anion channel VRAC (also referred to as VSOR, VSOAC) induced by activation of S1P receptors. We have previously reported about this S1P-induced ATP release measured by voltage clamp and luciferase assay. Here, we investigated whether the S1P-induced ATP release can affect cell functions like cell migration by activating purinergic P2X or P2Y receptors. The microglia cell line BV-2 has been used to conduct the experiments. In order to assess the effects of a S1P-induced ATP release we used scratch assays (also ‘wound healing assay’). S1P, like ATP and ADP, stimulates cell migration into the scratch area. The inhibition of S1P receptors and of the downstream G i proteins reduced the cell migration. Antagonists of VRAC, which lead to reduced ATP release, were also able to diminish the cell migration. Furthermore, direct inhibition of ATP-gated P2X4 or P2X7 receptors or ADP-stimulated P2Y12 receptors blocked the stimulating effects of S1P on cell migration. We conclude that there is an interaction between S1P receptors and purinergic receptors mediated by a S1P-induced ATP release via VRAC and that the amount of released ATP is capable to stimulate cell migration of BV-2 microglia cells via activation of P2X4, P2X7 and P2Y12 receptors.