Resolving dynamics of cell signaling via real-time imaging of the immunological synapse.

Stevens, Mark A. ; Pfeiffer, Janet R. (University of New Mexico, Albuquerque, NM) ; Wilson, Bridget S. (University of New Mexico, Albuquerque, NM) ; Timlin, Jerilyn Ann ; Thomas, James L. (University of New Mexico, Albuquerque, NM) ; Lidke, Keith A. (Universit

关键词: BACTERIA;    LIPIDS;    MACROPHAGES;    MAST CELLS;    MEMBRANES;    MICROSCOPY;    PROTEINS;    REFLECTION;    SIMULATION;    SPECTROSCOPY Microscopy;    Electron-instrumentation.;    Protein-analysis.;    Microscopy-Technique.;   

DOI  :  10.2172/974405
RP-ID  :  SAND2009-6396
PID  :  OSTI ID: 974405
Others  :  TRN: US201009%%70

美国|英语

来源: SciTech Connect

【 摘 要 】

This highly interdisciplinary team has developed dual-color, total internal reflection microscopy (TIRF-M) methods that enable us to optically detect and track in real time protein migration and clustering at membrane interfaces. By coupling TIRF-M with advanced analysis techniques (image correlation spectroscopy, single particle tracking) we have captured subtle changes in membrane organization that characterize immune responses. We have used this approach to elucidate the initial stages of cell activation in the IgE signaling network of mast cells and the Toll-like receptor (TLR-4) response in macrophages stimulated by bacteria. To help interpret these measurements, we have undertaken a computational modeling effort to connect the protein motion and lipid interactions. This work provides a deeper understanding of the initial stages of cellular response to external agents, including dynamics of interaction of key components in the signaling network at the 'immunological synapse,' the contact region of the cell and its adversary.