Vinculin knockout results in heart and brain defects during embryonic development.
Vinculin knockout results in heart and brain defects during embryonic development.
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发表时间：
1998-01
期刊：
影响因子：
4.6
通讯作者：
Weiming Xu;H. Baribault;E. Adamson
中科院分区：
文献类型：
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作者：
Weiming Xu;H. Baribault;E. Adamson
黏着斑蛋白基因编码一种细胞骨架蛋白，存在于粘着斑和细胞-细胞粘附连接处。通过在胚胎干（ES）细胞中使用靶向载体进行同源重组来灭活白蛋白。通过建立的程序将杂合ES细胞引入小鼠中以产生正常且可生育的杂合动物。没有纯合黏着斑蛋白-/-胚胎出生，妊娠期间的分析表明，黏着斑蛋白无效胚胎从E8天开始就很小且异常，但有些存活至E10。最突出的缺陷是缺乏中线融合的吻神经管，产生颅双叶外观和衰减的颅神经和脊神经发育。E9.5时心脏发育减少，心肌和内皮结构严重减少和无动力。突变胚胎体积缩小30-40%，体节和肢体发育迟缓，外胚层组织稀疏脆弱。与野生型水平相比，从突变胚胎中分离的成纤维细胞（MEF）对纤连蛋白、玻连蛋白、层粘连蛋白和胶原蛋白的粘附力降低。此外，在这些基质上的迁移率高出两倍，粘着斑激酶（FAK）活性水平高出三倍。我们的结论是黏着斑蛋白是必要的正常胚胎发育，可能是因为它的作用，在调节细胞粘附和运动，细胞行为的正常胚胎形态发生是必不可少的，虽然在神经和心脏发育的具体作用不能排除。
The vinculin gene codes for a cytoskeletal protein, found in focal adhesion plaques and in cell-cell adherens junctions. Vinculin was inactivated by homologous recombination using a targeting vector in embryonic stem (ES) cells. The heterozygous ES cells were introduced into mice by established procedures to produce heterozygous animals that were normal and fertile. No homozygous vinculin-/- embryos were born and analyses during the gestational period showed that the vinculin null embryos were small and abnormal from day E8 but some survived until E10. The most prominent defect was lack of midline fusion of the rostral neural tube, producing a cranial bilobular appearance and attenuation of cranial and spinal nerve development. Heart development was curtailed at E9.5, with severely reduced and akinetic myocardial and endocardial structures. Mutant embryos were 30-40% smaller, somites and limbs were retarded and ectodermal tissues were sparse and fragile. Fibroblasts (MEF) isolated from mutant embryos were shown to have reduced adhesion to fibronectin, vitronectin, laminin and collagen compared to wild-type levels. In addition, migration rates over these substrata were two-fold higher and the level of focal adhesion kinase (FAK) activity was three-fold higher. We conclude that vinculin is necessary for normal embryonic development, probably because of its role in the regulation of cell adhesion and locomotion, cell behaviors essential for normal embryonic morphogenesis, although specific roles in neural and cardiac development cannot be ruled out.