title: Analyzing actin dynamics during the activation of the B cell receptor in live B cells.
abstract: Actin reorganization has been shown to be important for lymphocyte activation in response to antigenic stimulation. However, methods for quantitative analysis of actin dynamics in live lymphocytes are still underdeveloped. In this study, we describe new methods to examine the actin dynamics in B cells induced by antigenic stimulation. Using the A20 B cell line expressing GFP-actin, we analyzed in real time the redistribution of F-actin and the lateral mobility of actin flow in the surface of B cells in response to soluble and/or membrane associated antigens. Using fluorescently labeled G-actin, we identified the subcellular location and quantified the level of de novo actin polymerization sites in primary B cells. Using A20 B cells expressing G-actin fused with the photoconvertible protein mEos, we examined the kinetics of actin polymerization and depolymerization at the same time. Our studies present a set of methods that are capable of quantitatively analyzing the role of actin dynamics in lymphocyte activation.
authors.affiliation: Department of Cell Biology & Molecular Genetics, University of Maryland, College Park, MD 20742, USA. cliu1234@umd.edu
authors.forename: Chaohong
authors.initials: C
authors.lastname: Liu
authors.forename: Heather
authors.initials: H
authors.lastname: Miller
authors.forename: Shruti
authors.initials: S
authors.lastname: Sharma
authors.forename: Amy
authors.initials: A
authors.lastname: Beaven
authors.forename: Arpita
authors.initials: A
authors.lastname: Upadhyaya
authors.forename: Wenxia
authors.initials: W
authors.lastname: Song
mesh_terms: D008841:Actin Cytoskeleton / Q000276:immunology; D000199:Actins / Q000378:metabolism*; D000818:Animals; D000941:Antigens / Q000276:immunology / Q000494:pharmacology; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002352:Carrier Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000276:immunology; D004268:DNA-Binding Proteins; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000378:metabolism; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D000199:Actins; D000941:Antigens; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D009419:Nerve Tissue Proteins; D011947:Receptors, Antigen, B-Cell; D011993:Recombinant Fusion Proteins; C118760:Zfpn1a4 protein, mouse; D049452:Green Fluorescent Proteins