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

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• 作者：Hannah Schachtner ; Ang Li ; David Stevenson ; Simon D.J. Calaminus ; Steven G. Thomas ; Steve P. Watson ; Michael Sixt ; Rol ; Wedlich-Soldner ; Douglas Strathdee ; Laura M. Machesky
• 关键词：Actin dynamics ; Cell migration ; Cytokinesis ; Motility ; Cytoskeleton ; Imaging
• 刊名：European Journal of Cell Biology
• 出版年：2012
• 出版时间：November-December, 2012
• 年：2012
• 卷：91
• 期：11-12
• 页码：923-929
• 全文大小：1312 K

文摘

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.