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

Schachtner, H., Li, A., Stevenson, D., Calaminus, S. D.J., Thomas, S. G., Watson, S. P., Sixt, M., Wedlich-Soldner, R., Strathdee, D. and Machesky, L. M. (2013) Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo. European Journal of Cell Biology, 91(11-12), pp. 923-929. (doi:10.1016/j.ejcb.2012.04.002)

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Publisher's URL: http://dx.doi.org/10.1016/j.ejcb.2012.04.002

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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Li, Mr Ang and Machesky, Professor Laura and Strathdee, Dr Douglas and Calaminus, Dr Simon and Stevenson, Dr David
Authors: Schachtner, H., Li, A., Stevenson, D., Calaminus, S. D.J., Thomas, S. G., Watson, S. P., Sixt, M., Wedlich-Soldner, R., Strathdee, D., and Machesky, L. M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
Journal Name:European Journal of Cell Biology
Publisher:Elsevier B.V.
ISSN:0171-9335
Published Online:31 May 2012
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
533601The role of podosomes and lamellipodia in megakaryocyte function.Laura MacheskyBritish Heart Foundation (BHF)FS/09/034/27756ICS - BEATSON INSTITUTE FOR CANCER RES.