|Detection of CD31/PECAM‑1 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Rat Anti-Mouse CD31/PECAM‑1 APC‑conjugated Monoclonal Antibody (Catalog # FAB6874A, filled histogram) or isotype control antibody (Catalog # IC005A, open histogram). View our protocol for Staining Membrane-associated Proteins.|
PECAM-1 (Platelet-Endothelial Cell Adhesion Molecule-1), also known as CD31, is a 130 kDa type I transmembrane glycoprotein adhesion molecule that belongs to the immunoglobulin superfamily (1, 2). Expression is restricted to cells involved in circulation, especially endothelial cells, platelets, monocytes, neutrophils and lymphocyte subsets. CD31 is concentrated at cell-cell junctions and is required for transendothelial migration (TEM) (1‑3). The extracellular domain (ECD) of CD31 has ten potential N-linked glycosylation sites and six C2-type Ig-like domains, the first of which is critical for adhesion and extravasation (3, 4). The cytoplasmic domain contains immunoregulatory tyrosine-based inhibitory and switch motifs (ITIM, ITSM) that mediate both inhibition and activation via phosphotyrosine-mediated engagement of SH2-containing signaling molecules (1, 5). Metalloproteinase-mediated ectodomain shedding occurs during apoptosis (6) but increased serum CD31 ectodomain in HIV and active multiple sclerosis occurs independent of apoptosis (7, 8). In humans, expression of six isoforms with exon deletions in the cytoplasmic domain is tissue- and stage-specific, but full-length CD31 is predominant. A form lacking the ITSM predominates in mouse (9). Mouse CD31 ECD shows 77%, 63%, 63%, 63% and 61% amino acid (aa) sequence identity with rat, human, canine, porcine and bovine CD31, respectively. Although, CD31 participates with other adhesion molecules in some functions, it is the critical molecule for TEM. Homotypic CD31 adhesion in trans, combined with cycling of CD31 to and from surface-connected endothelial cell vesicles, leads leukocytes across endothelial tight junctions (3, 10). Homotypic adhesion and signaling functions also strongly suppress mitochondria-dependent apoptosis (11). In platelets, CD31 is necessary for limiting thrombus formation and promoting integrin-mediated clot retraction and platelet spreading, but the mechanism for these phenomena is unclear (12-13). CD31-/- mice are deficient in chemokine-mediated chemotaxis (14).
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