Suzuki Akio S, Manabe Junichi, Hirakawa Asako
Department of Biology, Faculty of General Education, Kumamoto University, 2-40-1 Kurokami, 860, Kumamoto, Japan.
Rouxs Arch Dev Biol. 1991 Sep;200(4):213-222. doi: 10.1007/BF00361340.
For analysing spatial distribution of maternal proteins in an amphibian egg, monoclonal antibodies specific to certain regions were raised. One monoclonal antibody was found (MoAB Xa5B6) which reacted specifically with the animal hemisphere of the mature Xenopus laevis egg. The maternal protein that reacted with the MoAb Xa5B6 was shown to be distributed asymmetrically along the dorso-ventral axis in the upper region of the equatorial zone of the fertilized egg. At late blastula stage, the antigen protein could be observed clearly in both the marginal zone and animal cap. It was localized predominantly in mesodermal and ectodermal cells of late neurula embryos. The Xa5B6 antigen accumulated during oogenesis. The distribution pattern of maternal protein was remarkably different in the developmental stages of the oocyte. The pattern in the mature oocyte was completely different from that of the immature egg in which the antigen was located in the radial striations of the oocyte cytoplasm. After maturation, the distribution pattern changed drastically to an animal-vegetal polarization and the striation labellings were no longer observed. By Western blot examination, it was confirmed that the amounts of antigen protein were constant during early embryogenesis and the mesoectoderm contained a greater amount of antigens than the endoderm at late blastula. The antibody detected two bands of approximately 70 × 10 and 30 × 10 Mr by Western blot analysis. The latter molecule may possibly be a degrading moiety of the former. The results were discussed in relation to establishment of animal-vegetal (A/V) and dorso-ventral (D/V) polarization at the molecular level.
为了分析两栖类卵中母体蛋白的空间分布,制备了针对特定区域的单克隆抗体。发现了一种单克隆抗体(MoAB Xa5B6),它与成熟非洲爪蟾卵的动物半球发生特异性反应。与MoAb Xa5B6反应的母体蛋白在受精卵赤道区上部沿背腹轴呈不对称分布。在囊胚晚期,抗原蛋白在边缘区和动物帽中都能清晰观察到。它主要定位于神经胚晚期胚胎的中胚层和外胚层细胞中。Xa5B6抗原在卵子发生过程中积累。母体蛋白的分布模式在卵母细胞的发育阶段有显著差异。成熟卵母细胞中的模式与未成熟卵完全不同,未成熟卵中的抗原位于卵母细胞细胞质的放射状条纹中。成熟后,分布模式急剧变为动植物极化,不再观察到条纹标记。通过蛋白质免疫印迹检测证实,在早期胚胎发生过程中抗原蛋白的量是恒定的,在囊胚晚期中胚外胚层比内胚层含有更多的抗原。通过蛋白质免疫印迹分析,该抗体检测到两条分别约为70×10和30×10 Mr的条带。后一种分子可能是前一种分子的降解部分。结合在分子水平上动植物(A/V)和背腹(D/V)极化的建立对结果进行了讨论。