Lau Jeffrey M C, Muslin Anthony J
Center for Cardiovascular Research, Department of Medicine, Department of Cell Biology & Physiology, Washington University School of Medicine, St. Louis, MO, USA.
Methods Mol Biol. 2009;518:31-41. doi: 10.1007/978-1-59745-202-1_3.
The 14-3-3 intracellular phosphoserine/threonine-binding proteins are adapter molecules that regulate signal transduction, cell cycle, nutrient sensing, apoptotic, and cytoskeletal pathways. There are seven 14-3-3 family members, encoded by separate genes, in vertebrate organisms. To evaluate the role of individual 14-3-3 proteins in vertebrate embryonic development, we utilized an antisense morpholino oligo microinjection technique in Xenopus laevis embryos. By use of this method, we showed that embryos lacking specific 14-3-3 proteins displayed unique phenotypic abnormalities. Specifically, embryos lacking 14-3-3 tau exhibited gastrulation and axial patterning defects, but embryos lacking 14-3-3 gamma exhibited eye defects without other abnormalities, and embryos lacking 14-3-3 zeta appeared completely normal. These and other results demonstrate the power and specificity of the morpholino antisense oligo microinjection technique.
14-3-3细胞内磷酸丝氨酸/苏氨酸结合蛋白是调节信号转导、细胞周期、营养感知、凋亡和细胞骨架途径的衔接分子。在脊椎动物中，有七个由不同基因编码的14-3-3家族成员。为了评估单个14-3-3蛋白在脊椎动物胚胎发育中的作用，我们在非洲爪蟾胚胎中利用了反义吗啉代寡核苷酸显微注射技术。通过使用这种方法，我们发现缺乏特定14-3-3蛋白的胚胎表现出独特的表型异常。具体而言，缺乏14-3-3 tau的胚胎表现出原肠胚形成和轴向模式缺陷，但缺乏14-3-3 gamma的胚胎表现出眼部缺陷而无其他异常，而缺乏14-3-3 zeta的胚胎看起来完全正常。这些以及其他结果证明了吗啉代反义寡核苷酸显微注射技术的强大功能和特异性。
