Liu Zhicen, Nukazuka Akira, Takagi Shin
Center for Gene Research, Nagoya University, Furo-cho, Chikusa 464-8602, Japan.
Dev Growth Differ. 2007 Jan;49(1):49-59. doi: 10.1111/j.1440-169X.2007.00906.x.
The formation of intricate and functional biological structures depends on the dynamic changes of cellular morphology. Confocal laser scanning microscopy (CLSM) is a widely used method to reveal the three-dimensional (3-D) structure of cells during the development of Caenorhabditis elegans (C. elegans) and other model organisms. Improving the efficiency and image quality of CLSM would benefit studies using this method. We found that CED-10::GFP::CED-10, a green fluorescent protein (GFP) marker, is intensely expressed beneath the cell surface, facilitating visualization of cellular morphology in C. elegans embryos. By combining the unique properties of this marker, and with the help of direct 3-D rendering of images obtained by CLSM, we developed a simple but powerful method for investigating cellular morphology in developing embryos. Using this method we, for the first time, document the dynamic changes in the morphology of ventral neuroblasts in vivo during ventral cleft closure.
复杂且功能性生物结构的形成依赖于细胞形态的动态变化。共聚焦激光扫描显微镜(CLSM)是一种广泛应用的方法,用于揭示秀丽隐杆线虫(C. elegans)及其他模式生物发育过程中细胞的三维(3-D)结构。提高CLSM的效率和图像质量将有益于使用该方法的研究。我们发现,绿色荧光蛋白(GFP)标记物CED-10::GFP::CED-10在细胞表面下方强烈表达,有助于观察秀丽隐杆线虫胚胎中的细胞形态。通过结合该标记物的独特特性,并借助CLSM获取图像的直接三维渲染,我们开发了一种简单而强大的方法来研究发育中胚胎的细胞形态。使用这种方法,我们首次记录了腹侧裂隙闭合过程中体内腹侧神经母细胞形态的动态变化。