for the PIP2 messenger system produces two effectors, phospholipase C (PLC) and phosphoinositide 3-kinase (PI3K). PLC as an effector produces two different second messengers, inositol triphosphate (IP3) and Diacylglycerol (DAG). IP3 is soluble and diffuses freely into the cytoplasm. As a second messenger, it is recognized by the inositol triphosphate receptor (IP3R), a Ca2+ channel in the endoplasmic reticulum (ER) membrane, which stores intracellular Ca2+. The binding of IP3 to IP3R releases Ca2+ from the ER into the normally Ca2+-poor cytoplasm, which then triggers various events of Ca2+ signaling. Specifically in blood vessels, the increase in Ca2+ concentration from IP3 releases nitric oxide, which then diffuses into the smooth muscle tissue and causes relaxation. DAG remains bound to the membrane by its fatty acid "tails" where it recruits and activates both conventional and novel members of the protein kinase C family. Thus, both IP3 and DAG contribute to activation of PKCs. Phosphoinositide 3-kinase (PI3K) as an effector phosphorylates phosphatidylinositol bisphosphate (PIP2) to produce phosphatidylinositol (3,4,5)-trisphosphate (PIP3). PIP3 has been shown to activate protein kinase B, increase binding to extracellular proteins and ultimately enhance cell survival. Activators of G-protein coupled receptors See main article on G-protein coupled receptors Lysophosphatidic acid (LPA) LPA is the result of phospholipase A2 action on phosphatidic acid. The SN-1 position can contain either an ester bond or an ether bond, with ether LPA being found at elevated levels in certain cancers. LPA binds the high-affinity G-protein coupled receptors LPA1, LPA2, and LPA3 (also known as EDG2, EDG4, and EDG7, respectively). Sphingosine-1-phosphate (S1P) S1P is present at high concentrations in plasma and secreted locally at elevated concentrations at sites of inflammation. It is formed by the regulated phosphorylation of sphingosine. It acts through five dedicated high-affinity G-protein coupled receptors, S1P1 - S1P5. Targeted deletion of S1P1 results in lethality in mice and deletion of S1P2 results in seizures and deafness. Additionally, a mere 3- to 5-fold elevation in serum S1P concentrations induces sudden cardiac death by an S1P3-receptor specific mechanism. Platelet activating factor (PAF) PAF is a potent activator of platelet aggregation, inflammation, and anaphylaxis. It is similar to the ubiquitous membrane phospholipid phosphatidylcholine except that it contains an acetyl-group in the SN-2 position and the SN-1 position contains an ether-linkage. PAF signals through a dedicated G-protein coupled receptor, PAFR and is inactivated by PAF acetylhydrolase. Endocannabinoids The endogenous cannabinoids, or endocannabinoids, are endogenous lipids that activate