Document Detail

Quantifying subcellular distribution of fluorescent fusion proteins in cells migrating within tissues.
MedLine Citation:
PMID:  20956985     Owner:  NLM     Status:  In-Data-Review    
The movement of proteins within cells can provide dynamic indications of cell signaling and cell polarity, but methods are needed to track and quantify subcellular protein movement within tissue environments. Here we present a semiautomated approach to quantify subcellular protein location for hundreds of migrating cells within intact living tissue using retrovirally expressed fluorescent fusion proteins and time-lapse two-photon microscopy of intact thymic lobes. We have validated the method using GFP-PKCζ, a marker for cell polarity, and LAT-GFP, a marker for T-cell receptor signaling, and have related the asymmetric distribution of these proteins to the direction and speed of cell migration. These approaches could be readily adapted to other fluorescent fusion proteins, tissues and biological questions.
Heather J Melichar; Ou Li; Paul Herzmark; Raghav K Padmanabhan; Jane Oliaro; Mandy J Ludford-Menting; Philippe Bousso; Sarah M Russell; Badrinath Roysam; Ellen A Robey
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Publication Detail:
Type:  Journal Article     Date:  2010-10-19
Journal Detail:
Title:  Immunology and cell biology     Volume:  89     ISSN:  1440-1711     ISO Abbreviation:  Immunol. Cell Biol.     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-05-06     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8706300     Medline TA:  Immunol Cell Biol     Country:  England    
Other Details:
Languages:  eng     Pagination:  549-57     Citation Subset:  IM    
Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
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