The Rac-FRET Mouse Reveals Tight Spatiotemporal Control of Rac Activity in Primary Cells and Tissues

The small G protein family Rac has numerous regulators that integrate extracellular signals into tight spatiotemporal maps of its activity to promote specific cell morphologies and responses. Here, we have generated a mouse strain, Rac-FRET, which ubiquitously expresses the Raichu-Rac biosensor. It...

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Published in:Cell reports (Cambridge) Vol. 6; no. 6; pp. 1153 - 1164
Main Authors: Johnsson, Anna-Karin E., Dai, Yanfeng, Nobis, Max, Baker, Martin J., McGhee, Ewan J., Walker, Simon, Schwarz, Juliane P., Kadir, Shereen, Morton, Jennifer P., Myant, Kevin B., Huels, David J., Segonds-Pichon, Anne, Sansom, Owen J., Anderson, Kurt I., Timpson, Paul, Welch, Heidi C.E.
Format: Journal Article
Language:English
Published: United States Cell Press 01.03.2014
Elsevier
Subjects:
Animals
Enzyme Activation
Fluorescence Resonance Energy Transfer - methods
Mice
Neutrophils - cytology
Neutrophils - enzymology
rac GTP-Binding Proteins - chemistry
rac GTP-Binding Proteins - metabolism
Resource
Signal Transduction
Spatio-Temporal Analysis
ISSN:2211-1247, 2211-1247
Online Access:Get full text
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Summary:The small G protein family Rac has numerous regulators that integrate extracellular signals into tight spatiotemporal maps of its activity to promote specific cell morphologies and responses. Here, we have generated a mouse strain, Rac-FRET, which ubiquitously expresses the Raichu-Rac biosensor. It enables FRET imaging and quantification of Rac activity in live tissues and primary cells without affecting cell properties and responses. We assessed Rac activity in chemotaxing Rac-FRET neutrophils and found enrichment in leading-edge protrusions and unexpected longitudinal shifts and oscillations during protruding and stalling phases of migration. We monitored Rac activity in normal or disease states of intestinal, liver, mammary, pancreatic, and skin tissue, in response to stimulation or inhibition and upon genetic manipulation of upstream regulators, revealing unexpected insights into Rac signaling during disease development. The Rac-FRET strain is a resource that promises to fundamentally advance our understanding of Rac-dependent responses in primary cells and native environments.
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Present address: Research Centre for Animal Genetic Resources of the Mongolia Plateau, Inner Mongolia University, 235 West University Road, 010021 Hohhot, China
These authors contributed equally to this work
ISSN:2211-1247
2211-1247
DOI:10.1016/j.celrep.2014.02.024