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Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo

Calaminus, Simon D. J.; Calaminus, Simon D J; Calaminus, Simon; Schachtner, Hannah; Li, Ang; Stevenson, David; Calaminus, Simon D.J.; Thomas, Steven G.; Watson, Steve P.; Sixt, Michael; Wedlich-Soldner, Roland; Strathdee, Douglas; Machesky, Laura M.

Authors

Simon D. J. Calaminus

Simon D J Calaminus

Simon Calaminus S.Calaminus@hull.ac.uk

Hannah Schachtner

Ang Li

David Stevenson

Simon D.J. Calaminus

Steven G. Thomas

Steve P. Watson

Michael Sixt

Roland Wedlich-Soldner

Douglas Strathdee

Laura M. Machesky



Abstract

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.

Citation

Schachtner, H., Li, A., Stevenson, D., Calaminus, S. D., Thomas, S. G., Watson, S. P., …Machesky, L. M. (2012). Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo. European journal of cell biology, 91(11-12), (923-929). doi:10.1016/j.ejcb.2012.04.002. ISSN 0171-9335

Journal Article Type Article
Acceptance Date Apr 12, 2012
Online Publication Date May 31, 2012
Publication Date 2012-11
Publicly Available Date
Print ISSN 0171-9335
Electronic ISSN 1618-1298
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 91
Issue 11-12
Pages 923-929
DOI https://doi.org/10.1016/j.ejcb.2012.04.002
Keywords Actin dynamics, cell migration, cytokinesis, motility, cytoskeleton, imaging
Public URL https://hull-repository.worktribe.com/output/428945