Chemokines have a range of functions, including the activation and promotion of the vectorial migration of leukocytes. They mediate their biological effects by binding to their cognate G-protein-coupled receptors. Upon activation of the heterotrimeric G proteins, the Gα subunit exchanges GDP for GTP and dissociates from the receptor and from the Gβγ subunits, and both G-protein complexes go on to activate other downstream signaling events. In addition, chemokines interact with cell-surface glycosaminoglycans (GAGs). This potential for binding GAG components of proteoglycans on the cell surface or within the extracellular matrix allows the formation of the stable chemokine gradients necessary for leukocyte chemotaxis. In this chapter, we describe techniques for studying chemotaxis both in vivo and in vitro, as well as the creation of chemokine receptor-expressing cell lines, in order to examine this process in isolation.
Keywords: Air pouch; Chemokine; Chemokine receptor; Chemotactic gradient; Diffusion gradient chemotaxis; Glycosaminoglycans; In vivo chemotaxis; Stable transfectants; Transendothelial chemotaxis.
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