Chemokine receptor binding and signal transduction in native cells of the central nervous system.
Chemokine receptors belong to the superfamily of seven-transmembrane-spanning, G-protein-coupled receptors, and their expression by central nervous system cells is clearly documented. As this gene family has become the target of novel therapeutic development, the analysis of these receptors requires radioligand binding techniques as well as methods that entail assessing receptor stimulation of signal transduction pathways. Herein, we describe specific protocols for measuring radiolabeled chemokine binding to their cognate receptors on cultured glial cells as well as to receptors expressed in heterologous cell systems. Multiple downstream signaling pathways, including intracellular calcium influx and receptor-dependent kinase activation, are associated with chemokine receptor stimulation. Protocols for measuring these signaling events in chemokine-receptor-expressing cells are also presented.
Department of Pharmacology & Therapeutics, University of Florida College of Medicine, Gainesville, FL 32610-0267, USA.
Christopher N
CN
Davis
Shuzhen
S
Chen
Stefen A
SA
Boehme
Kevin B
KB
Bacon
Jeffrey K
JK
Harrison
D000818:Animals; D001671:Biochemistry / Q000379:methods*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002490:Central Nervous System / Q000378:metabolism*; D018925:Chemokines / Q000378:metabolism; D020935:MAP Kinase Signaling System; D017628:Microglia / Q000378:metabolism; D008954:Models, Biological; D011233:Precipitin Tests; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D051381:Rats; D017207:Rats, Sprague-Dawley; D019707:Receptors, Chemokine / Q000378:metabolism*; D015398:Signal Transduction
D018925:Chemokines; D011518:Proto-Oncogene Proteins; D019707:Receptors, Chemokine; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D002118:Calcium