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Spatiotemporal resolution of Ca 2+ signaling events by real time imaging of single B cells
Author(s) -
Junek Stephan,
Engelke Michael,
Schild Detlev,
Wienands Jürgen
Publication year - 2012
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2012.03.057
Subject(s) - microbiology and biotechnology , cytosol , signal transduction , effector , organelle , signal transducing adaptor protein , intracellular , second messenger system , live cell imaging , cell , subcellular localization , cell signaling , receptor , chemistry , cytoplasm , biology , biochemistry , enzyme
Antigen‐induced B cell activation requires mobilization of the Ca 2+ second messenger. This process is associated with the subcellular relocalization of signal effector proteins of the B cell antigen receptor such as the adaptor protein SLP65. Here we describe a broadly applicable live cell imaging method to simultaneously visualize intracellular Ca 2+ flux profiles and the translocation of cytosolic signaling proteins to the plasma membrane in real time. Our approach delineated the kinetic hierarchy of Ca 2+ signaling events in B cells and revealed a timely ordered contribution of various organelles to the overall Ca 2+ signal. The developed experimental setup provides a useful tool to resolve the spatiotemporal signaling dynamics in various receptor signaling systems.