A study of wound repair in Dictyostelium cells by using novel laserporation.
A study of wound repair in Dictyostelium cells by using novel laserporation.
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DOI：
10.1038/s41598-018-26337-0
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发表时间：
2018-05-22
影响因子：
4.6
通讯作者：
Yumura S
中科院分区：
文献类型：
--
作者：
Pervin MS;Itoh G;Talukder MSU;Fujimoto K;Morimoto YV;Tanaka M;Ueda M;Yumura S
我们用一种新的基于激光的细胞穿孔方法研究了网柄苔藓细胞的细胞膜修复机制。通过荧光细胞膜蛋白的实时成像、荧光染料的内流和钙离子成像来表征创面毛孔的张开和闭合的动态。根据伤口的大小，伤口在2-4 秒内闭合。细胞能耐受小于2.0 µm的伤口，在无钙离子存在的情况下，伤口毛孔不闭合，细胞破裂。细胞内钙离子的释放也促进了细胞内钙离子的升高，但对创面修复无影响。依赖于外界的钙离子浓度，膜联蛋白C1立即在伤口处积累，而膜联蛋白C1敲除细胞在伤口修复过程中存在缺陷，但不是必需的。Dictyostelius细胞能够以相同的时间进程对多个重复创伤做出反应，这与以前的报道相反，即第一个创伤加速了成纤维细胞的第二个伤口修复。
We examined the mechanism of cell membrane repair in Dictyostelium cells by using a novel laser-based cell poration method. The dynamics of wound pores opening and closing were characterized by live imaging of fluorescent cell membrane proteins, influx of fluorescent dye, and Ca2+ imaging. The wound closed within 2–4 sec, depending on the wound size. Cells could tolerate a wound size of less than 2.0 µm. In the absence of Ca2+ in the external medium, the wound pore did not close and cells ruptured. The release of Ca2+ from intracellular stores also contributed to the elevation of cytoplasmic Ca2+ but not to wound repair. Annexin C1 immediately accumulated at the wound site depending on the external Ca2+ concentration, and annexin C1 knockout cells had a defect in wound repair, but it was not essential. Dictyostelium cells were able to respond to multiple repeated wounds with the same time courses, in contrast to previous reports showing that the first wound accelerates the second wound repair in fibroblasts.
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