Ultra-high-throughput Ca²⁺ assay in platelets to distinguish ITAM-linked and G-protein-coupled receptor activation.

ABSTRACT: Antiplatelet drugs targeting G-protein-coupled receptors (GPCRs), used for the secondary prevention of arterial thrombosis, coincide with an increased bleeding risk. Targeting ITAM-linked receptors, such as the collagen receptor glycoprotein VI (GPVI), is expected to provide a better antithrombotic-hemostatic profile. Here, we developed and characterized an ultra-high-throughput (UHT) method based on intracellular [Ca²⁺]_i increases to differentiate GPVI and GPCR effects on platelets. In 96-, 384-, or 1,536-well formats, Calcium-6-loaded human platelets displayed a slow-prolonged or fast-transient [Ca²⁺]_i increase when stimulated with the GPVI agonist collagen-related peptide or with thrombin and other GPCR agonists, respectively. Semi-automated c

SUBMITTER: Fernandez DI 

PROVIDER: S-EPMC8762394 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature