Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo.
We describe here the development and characterization of a conditionally inducible mouse model expressing Lifeact-GFP, a peptide that reports the dynamics of filamentous actin. We have used this model to study platelets, megakaryocytes and melanoblasts and we provide evidence that Lifeact-GFP is a useful reporter in these cell types ex vivo. In the case of platelets and megakaryocytes, these cells are not transfectable by traditional methods, so conditional activation of Lifeact allows the study of actin dynamics in these cells live. We studied melanoblasts in native skin explants from embryos, allowing the visualization of live actin dynamics during cytokinesis and migration. Our study revealed that melanoblasts lacking the small GTPase Rac1 show a delay in the formation of new pseudopodia following cytokinesis that accounts for the previously reported cytokinesis delay in these cells. Thus, through use of this mouse model, we were able to gain insights into the actin dynamics of cells that could only previously be studied using fixed specimens or following isolation from their native tissue environment.
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD.
Hannah
H
Schachtner
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD.
Ang
A
Li
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD.
David
D
Stevenson
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD.
Simon D J
SDJ
Calaminus
Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT.
Steve
S
Thomas
Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT.
Steve P
SP
Watson
Institute of Science and Technology, Am Campus 1, A-3400 Klosterneuberg, Austria.
Michael
M
Sixt
Cellular Dynamics and Cell Patterning, Max-Planck Institute of Biochemistry, Am, Klopferspitz 18, 82152 Martinsried, Germany.
Roland
R
Wedlich-Soldner
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD.
Douglas
D
Strathdee
The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD.
Laura M
LM
Machesky
D008841:Actin Cytoskeleton / Q000648:ultrastructure*; D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D048749:Cytokinesis; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D008544:Melanocytes / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D058953:Neural Stem Cells / Q000378:metabolism / Q000648:ultrastructure; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity; D010455:Peptides / Q000235:genetics; D011554:Pseudopodia / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D059008:Time-Lapse Imaging; D014158:Transcription, Genetic; D020744:rac GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020830:rac1 GTP-Binding Protein
D000199:Actins; D009479:Neuropeptides; D010455:Peptides; C484602:Rac1 protein, mouse; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D020744:rac GTP-Binding Proteins; D020830:rac1 GTP-Binding Protein