NAADP/SERCA3-Dependent Ca²⁺ Stores Pathway Specifically Controls Early Autocrine ADP Secretion Potentiating Platelet Activation.

Abstract

Ca²⁺ signaling is a key and ubiquitous actor of cell organization and its modulation controls many cellular responses. SERCAs (sarco-endoplasmic reticulum Ca²⁺-ATPases) pump Ca²⁺ into internal stores that play a major role in the cytosolic Ca²⁺ concentration rise upon cell activation. Platelets exhibit 2 types of SERCAs, SERCA2b and SERCA3 (SERCA3 deficient mice), which may exert specific roles, yet ill-defined. We have recently shown that Ca²⁺ mobilization from SERCA3-dependent stores was required for full platelet activation in weak stimulation conditions. To uncover the signaling mechanisms associated with Ca²⁺ mobilization from SERCA3-dependent stores leading to ADP secretion. Using platelets from wild-type or <i>Serca3</i>-deficient mice, we demonstrated that an early (within 5-10 s following stimulation) secretion of ADP specifically dependent on SERCA3 stored Ca²⁺ is exclusively mobilized by nicotinic acid adenosine dinucleotide-phosphate (NAADP): both Ca²⁺ mobilization from SERCA3-dependent stores and primary ADP secretion are blocked by the NAADP receptor antagonist Ned-19, and reciprocally both are stimulated by permeant NAADP. In contrast, Ca²⁺ mobilization from SERCA3-dependent stores and primary ADP secretion were unaffected by inhibition of the production of IP3 (inositol-1,4,5-trisphosphate) by phospholipase-C and accordingly were not stimulated by permeant IP3. Upon activation, an NAADP/SERCA3 Ca²⁺ mobilization pathway initiates an early ADP secretion, potentiating platelet activation, and a secondary wave of ADP secretion driven by both an IP3/SERCA2b-dependent Ca²⁺ stores pathway and the NAADP/SERCA3 pathway. This does not exclude that Ca²⁺ mobilized from SERCA3 stores may also enhance platelet global reactivity to agonists. Because of its modulating effect on platelet activation, this NAADP-SERCA3 pathway may be a relevant target for anti-thrombotic therapy. Graphic Abstract: A graphic abstract is available for this article.