Spatiotemporal resolution of Ca2+ signaling events by real time imaging of single B cells.
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.
Department of Neurophysiology and Biophysics, Georg-August-University of Göttingen, Göttingen, Germany.
Stephan
S
Junek
Michael
M
Engelke
Detlev
D
Schild
Jürgen
J
Wienands
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D002451:Cell Compartmentation; D002460:Cell Line; D002645:Chickens; D018613:Microscopy, Confocal; D021381:Protein Transport; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism; D059010:Single-Cell Analysis
D048868:Adaptor Proteins, Signal Transducing; C113939:B cell linker protein; D011947:Receptors, Antigen, B-Cell; D002118:Calcium