Development of Mechanical Stability in Late-Stage Embryonic Erythroid Cells: Insights From Fluorescence Imaged Micro-Deformation Studies.
Development of Mechanical Stability in Late-Stage Embryonic Erythroid Cells: Insights From Fluorescence Imaged Micro-Deformation Studies.
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DOI：
10.3389/fphys.2021.761936
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发表时间：
2021
影响因子：
4
通讯作者：
Waugh RE
中科院分区：
文献类型：
--
作者：
Delgadillo LF;Huang YS;Leon S;Palis J;Waugh RE
荧光标记和红细胞显微操作的结合使用已被证明是理解和表征细胞力学行为的基本机制的有力工具。本研究采用该方法研究了原代胚胎红细胞中膜相关细胞骨架（MAS）的发育。红细胞生成在哺乳动物胚胎中有两种形式，原始和最终，分别以血管内和血管外成熟为特征。小鼠胚胎中的原始红系前体首先在胚胎日(E) 8.25开始循环，在E12.5和E16.5之间去核之前作为半同步队列成熟。先前，我们确定MAS的主要成分在E10.5和E12.5之间定位到膜上，并且这种定位与同一时期膜机械稳定性的增加有关。机械稳定性的变化反映在当细胞被吸进微移液管时形成的突起尖端膜的无mas区域的产生上。通过E14.5，在所有可检测到的细胞骨架成分定位到膜上至少2天后，原始红细胞继续成熟，形成无mas区域的倾向减少，这表明细胞骨架成分在膜定位后，膜内聚继续加强。在这里，我们证明了无MAS区域的形成是MAS内部机械故障的结果，而不是膜双分子层从MAS分离的结果。一旦MAS形成“孔”，骨骼网络沿吸入投影向外侧收缩，形成无MAS区域。在蛋白4.1缺失的原始红细胞中，形成无mas区域的倾向明显增强。值得注意的是，在来自人骨髓的成熟红细胞中也观察到类似的无mas区域，这表明终代红细胞中也发生了类似的过程。我们得出的结论是，在成熟过程中，细胞骨架成分定位于哺乳动物红细胞的细胞膜本身不足以产生成熟的MAS，但随后的过程还需要加强骨骼内的相互作用。
The combined use of fluorescence labeling and micro-manipulation of red blood cells has proven to be a powerful tool for understanding and characterizing fundamental mechanisms underlying the mechanical behavior of cells. Here we used this approach to study the development of the membrane-associated cytoskeleton (MAS) in primary embryonic erythroid cells. Erythropoiesis comes in two forms in the mammalian embryo, primitive and definitive, characterized by intra- and extra-vascular maturation, respectively. Primitive erythroid precursors in the murine embryo first begin to circulate at embryonic day (E) 8.25 and mature as a semi-synchronous cohort before enucleating between E12.5 and E16.5. Previously, we determined that the major components of the MAS become localized to the membrane between E10.5 and E12.5, and that this localization is associated with an increase in membrane mechanical stability over this same period. The change in mechanical stability was reflected in the creation of MAS-free regions of the membrane at the tips of the projections formed when cells were aspirated into micropipettes. The tendency to form MAS-free regions decreases as primitive erythroid cells continue to mature through E14.5, at least 2 days after all detectable cytoskeletal components are localized to the membrane, indicating continued strengthening of membrane cohesion after membrane localization of cytoskeletal components. Here we demonstrate that the formation of MAS-free regions is the result of a mechanical failure within the MAS, and not the detachment of membrane bilayer from the MAS. Once a “hole” is formed in the MAS, the skeletal network contracts laterally along the aspirated projection to form the MAS-free region. In protein 4.1-null primitive erythroid cells, the tendency to form MAS-free regions is markedly enhanced. Of note, similar MAS-free regions were observed in maturing erythroid cells from human marrow, indicating that similar processes occur in definitive erythroid cells. We conclude that localization of cytoskeletal components to the cell membrane of mammalian erythroid cells during maturation is insufficient by itself to produce a mature MAS, but that subsequent processes are additionally required to strengthen intraskeletal interactions.
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DOI：
10.1016/j.msec.2005.08.020
发表时间：
2006-09-01
期刊：
MATERIALS SCIENCE & ENGINEERING C-BIOMIMETIC AND SUPRAMOLECULAR SYSTEMS
影响因子：
--
作者：
Dao, M.;Li, J.;Suresh, S.
通讯作者：
Suresh, S.
影响因子：
20.3
作者：
Dahl, KN;Westhoff, CM;Discher, DE
通讯作者：
Discher, DE
影响因子：
7.5
作者：
Gautier, Emilie-Fleur;Leduc, Marjorie;Mayeux, Patrick
通讯作者：
Mayeux, Patrick
影响因子：
20.3
作者：
Lee, JCM;Gimm, JA;Chasis, JA
通讯作者：
Chasis, JA
影响因子：
2.9
作者：
EDER, PS;SOONG, CJ;TAO, M
通讯作者：
TAO, M