Extracellular ATP activates store-operated Ca2+ entry in white adipocytes: functional evidence for STIM1 and ORAI1

Here we have applied ratiometric measurements of intracellular Ca concentrations ([Ca ] ) to show that extracellularly applied ATP (100 mM) stimulates store-operated Ca entry (SOCE) in 3T3-L1 adipocytes. ATP produced a rapid increase in [Ca ] consisting of an initial transient elevation followed by a sustained elevated phase that could be observed only in the presence of extracellular Ca Gene expression data and [Ca ] recordings with uridine-5'-triphosphate (UTP) or with the phospholipase C (PLC) inhibitor U73122 demonstrated the involvement of purinergic P2Y2 receptors and the PLC/inositol trisphosphate (IP ) pathway. The [Ca ] elevation produced by re-introduction of a Ca -containing intracellular solution to adipocytes exposed to ATP in the absence of Ca was diminished by known SOCE antagonists. The chief molecular components of SOCE, the stromal interaction molecule 1 (STIM1) and the calcium release-activated calcium channel protein 1 (ORAI1), were detected at the mRNA and protein level. Moreover, SOCE was largely diminished in cells where STIM1 and/or ORAI1 had been silenced by small interfering (si)RNA. We conclude that extracellular ATP activates SOCE in white adipocytes, an effect predominantly mediated by STIM1 and ORAI1.