Objective: To investigate the regulatory effect of zyxin on the distribution of platelet cytoskeleton.
Methods: Platelets were isolated from zyxin-knockout (Zyx-/-) and wild type (WT) mice respectively and corresponding platelet cytoskeleton components were separated. The expressions of β-actin, α-actinin, filamin A and myosin ⅡA in cytoskeleton components were detected by Western blot. Actin polymerization was induced by the actin polymerization inducer Jasplakinolide (Jas) in WT and Zyx-/- platelets. Also, the expressions of the cytoskeleton proteins in cytoskeleton components were detected by Western blot. WT and Zyx-/- platelets were allowed to spread on fibrinogen-coated surface. Platelet F-actin was labeled with Alexa Fluor 488-conjugated phalloidin and the fluorescent intensity was compared between WT and Zyx-/- platelets.
Results: After zyxin gene was knockout, the expressions of cytoskeleton proteins β-actin, α-actinin, filamin A, and myosin Ⅱ A in resting and Jas-induced platelets were significantly increased. In the platelet spreading on fibrinogen surface, F-actin was increased in Zyx-/- platelets as compared with that in WT platelets.
Conclusion: Zyxin significantly regulates the distribution of platelet cytoskeleton, which plays an important role in maintaining platelet cytoskeleton homeostasis.
题目: Zyxin在调节血小板细胞骨架蛋白分布中的作用.
目的: 探讨斑联蛋白（zyxin）对血小板细胞骨架分布的调节作用.
方法: 分别取zyxin基因敲除（Zyx-/-）和野生型（WT）小鼠的血小板，分离血小板细胞骨架组分，应用Western blot分别检测细胞骨架组分中β-actin、α-actinin、filamin A及myosin ⅡA的表达；肌动蛋白聚合诱导剂（Jasplakinolide，Jas）诱导血小板肌动蛋白聚合，同法检测WT和Zyx-/-小鼠血小板细胞骨架组分中相关骨架蛋白的表达；将WT和Zyx-/-小鼠血小板于纤维蛋白原表面进行铺展，用Alexa Fluor 488偶联的鬼笔环肽标记肌动蛋白丝，比较WT和Zyx-/-小鼠血小板荧光强度.
结果: Zyxin基因敲除后，血小板主要细胞骨架蛋白β-actin、α-actinin、filamin A及myosin ⅡA在静息以及Jas诱导的血小板细胞骨架组分中的含量较WT小鼠血小板明显增多。在纤维蛋白原表面铺展的血小板中，Zyx-/-小鼠血小板肌动蛋白丝含量较WT小鼠血小板增多.
结论: Zyxin显著调节血小板细胞骨架分布，在维持血小板细胞骨架稳态中起着重要作用.