Block the function of nonmuscle myosin II by blebbistatin induces zebrafish embryo cardia bifida
Block the function of nonmuscle myosin II by blebbistatin induces zebrafish embryo cardia bifida
复制标题
肌球蛋白II阻断非肌肉肌球蛋白II的功能诱导斑马鱼胚胎贲门裂
DOI：
10.1007/s11626-014-9836-0
复制
发表时间：
2015
影响因子：
2.1
通讯作者：
Liu Dong
中科院分区：
文献类型：
--
作者：
Wang Xueqian;Chong Mei;Wang Xin;Wang Hongkui;Zhang Jie;Xu Hui;Zhang Jingjing;Liu Dong
非肌肉肌球蛋白II（NM II）是编码不同非肌肉肌球蛋白重链（包括NM II-A、NM II-B和NM II-C）的三种基因的多亚基蛋白产物的名称。Blebbistatin是一种小分子，已被证明是NM II的相对特异性抑制剂。阻断NM Ⅱ功能可诱导斑马鱼胚胎心裂，且呈剂量依赖性。用心室肌球蛋白重链（vmhc）和心房肌球蛋白重链（amhc）进行原位杂交分析，发现每颗心脏都含有不同的心房和心室。然而，心裂胚胎有高度可变的两个单独的心室之间的距离。我们还提供了证据表明，受精后12 - 20小时（hpf）的时间窗对于blebbistatin治疗引起的心裂形成是必要和充分的。在blebbistatin处理的胚胎中，spns 2的表达降低，表明NM II抑制引起的心裂表型与心前中胚层迁移缺陷有关。通过原位杂交分析，我们发现foxa 1在blebbistatin处理的24-hpf期胚胎的内胚层中表达，提示blebbistatin处理的贲门裂胚胎内胚层形成正常。此外，我们证明了blebbistatin处理导致斑马鱼心肌细胞在体内和新生小鼠心肌细胞在体外的形态学改变。
Nonmuscle myosin II (NM II) is the name given to the multi-subunit protein product of three genes encoding different nonmuscle myosin heavy chains including NM II-A, NM II-B, and NM II-C. Blebbistatin is a small molecule that has been shown to be a relatively specific inhibitor of NM II. Blocking the function of NM II by blebbistatin induces zebrafish embryo cardia bifida at a dose-dependent manner. In situ hybridization analysis with ventricular markerventricular myosin heavy chain(vmhc) and atrial markeratrial myosin heavy chain(amhc) showed each of the heart contained both distinct atria and ventricle. However, the cardia bifida embryos had highly variable distance between two separate ventricles. We also provided evidence that time window from 12 to 20 h post fertilization (hpf) is necessary and sufficient for cardia bifida formation caused by blebbistatin treatment. Expression ofspinster homolog 2(spns2) was decreased in blebbistatin-treated embryos, suggesting the cardia bifida phenotype caused by NM II inhibition was relevant to precardiac mesoderm migration defects. Through in situ hybridization analysis, we showed thatfoxa1was expressed in endoderm of blebbistatin-treated embryos at 24-hpf stage, suggesting the endoderm formation is normal in cardia bifida embryos caused by blebbistatin treatment. In addition, we demonstrated that blebbistatin treatment resulted in morphology alteration of zebrafish cardiomyocytes in vivo and neonatal mouse cardiomyocytes in vitro.