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

Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo.
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DOI:
10.1016/j.ejcb.2012.04.002
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发表时间:
2012-11
影响因子:
6.6
通讯作者:
Machesky, Laura M.
Machesky, Laura M.
中科院分区:
生物学3区
文献类型:
--
作者:
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.

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我们在这里描述了表达 Lifeact-GFP 的条件诱导小鼠模型的开发和表征,Lifeact-GFP 是一种报告丝状肌动蛋白动态的肽。我们已经使用该模型来研究血小板、巨核细胞和黑素细胞,并且我们提供的证据表明 Lifeact-GFP 是体外这些细胞类型中有用的报告基因。对于血小板和巨核细胞,这些细胞无法通过传统方法转染,因此 Lifeact 的条件激活可以实时研究这些细胞中的肌动蛋白动态。我们研究了来自胚胎的天然皮肤外植体中的黑素细胞,从而可以在胞质分裂和迁移过程中观察活肌动蛋白动态。我们的研究表明,缺乏小 GTPase Rac1 的成黑细胞在胞质分裂后显示出新伪足形成的延迟,这解释了先前报道的这些细胞中胞质分裂延迟的原因。因此,通过使用这种小鼠模型,我们能够深入了解细胞的肌动蛋白动力学,而这些细胞以前只能使用固定样本或从其天然组​​织环境中分离出来进行研究。
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.
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