Ezrin is a Major Regulator of Membrane Tension in Epithelial Cells.
Ezrin is a Major Regulator of Membrane Tension in Epithelial Cells.
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DOI：
10.1038/srep14700
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发表时间：
2015-10-05
影响因子：
4.6
通讯作者：
Janshoff A
中科院分区：
文献类型：
--
作者：
Rouven Brückner B;Pietuch A;Nehls S;Rother J;Janshoff A
质膜张力负责多种细胞功能，如运动性、细胞分裂和内吞作用。由于膜张力是由肌动蛋白皮质的质膜的内小叶的附件为主，我们调查的重要性埃兹林，一个主要的交联剂的膜-细胞骨架界面，汇合MDCK II细胞的细胞力学。为了这个目的，我们进行了埃兹蛋白耗尽实验，也提高了活跃埃兹蛋白分子在界面上的数量。力学性能进行了评估，通过力压痕实验，然后通过膜系绳提取。将PIP 2胶束注射到单个活细胞中以加强质膜和肌动蛋白-皮质之间的联系，同时通过ezrin siRNA和分别施用抑制剂新霉素和NSC 668394来减弱这种联系。我们观察到大量硬化的细胞和增加后，加入PIP 2胶束的膜张力。与此相反，减少活性ezrin导致膜张力的减少，伴随着多余的表面积的损失，增加皮质张力，肌动蛋白细胞骨架的重塑，和细胞高度的降低。这些数据证实了ezrin介导的质膜和皮质之间的连接对细胞力学和细胞形态的重要性。
Plasma membrane tension is responsible for a variety of cellular functions such as motility, cell division, and endocytosis. Since membrane tension is dominated by the attachment of the actin cortex to the inner leaflet of the plasma membrane, we investigated the importance of ezrin, a major cross-linker of the membrane-cytoskeleton interface, for cellular mechanics of confluent MDCK II cells. For this purpose, we carried out ezrin depletion experiments and also enhanced the number of active ezrin molecules at the interface. Mechanical properties were assessed by force indentation experiments followed by membrane tether extraction. PIP2 micelles were injected into individual living cells to reinforce the linkage between plasma membrane and actin-cortex, while weakening of this connection was reached by ezrin siRNA and administration of the inhibitors neomycin and NSC 668394, respectively. We observed substantial stiffening of cells and an increase in membrane tension after addition of PIP2 micelles. In contrast, reduction of active ezrin led to a decrease of membrane tension accompanied by loss of excess surface area, increase in cortical tension, remodelling of actin cytoskeleton, and reduction of cell height. The data confirm the importance of the ezrin-mediated connection between plasma membrane and cortex for cellular mechanics and cell morphology.