Studies on functional proteins for morphogenesis in early development of Xenopus laevis.

制备了针对不含卵黄蛋白的早期卵母细胞细胞质中抗原的单克隆抗体。另一种单克隆抗体采用成熟卵105000 × g上清液作为抗原制备。通过对卵巢组织切片的免疫染色，筛选出特别感兴趣的抗体。用不同发育阶段的卵母细胞染色，获得发育阶段特异性的10个单克隆抗体。对卵母细胞切片进行免疫染色，筛选出22种定位特异性抗体。定位特异性抗体分为动物半球细胞质（A型）、植物半球细胞质（V型）、卵母细胞外周细胞质（P型）和核质（N型）4类。在卵母细胞发育过程中，通过免疫组织化学方法在卵母细胞内同一区域检测到P型和N型抗体。根据Nieuwkoop和Faber的正常表，A型和V型的分布仅在发育阶段IV之后才在动物或植物半球发现。A型和V型的三维研究表明，它们可分为A_1型和A_2型，V_1型和V_2型。这些分布也被从成熟卵子的动物和植物半球提取的蛋白质的western-blotting分析所证实。部分A、V、P型抗体在受精卵胚期的分布进行了研究。它们的定位与成熟的卵子完全相同。

Monoclonal antibodies were prepared against antigens which are contained in cytoplasm of early stage oocytes without yolk protein. Another monoclonal antibodies were produced using 105,000xG super-natant of matured egg as antigens. Specially interesting antibodies were selected by the immunological staining on the histological sections of ovary. Developmental stage specific 10 monoclonal anti-bodies were obtained by staining test with oocytes of different developmental stages. Localization specific 22 antibodies were selected by the immunological staining on the sections of oocytes.Localization specific antibodies were classified into 4 groups such as cytoplasm of animal hemisphere (type A), cytoplasm of vegetal hemisphere (type V), peripheral cytoplasm of oocytes (type P) and nuclear plasm (type N). Antibodies of type P and N are detected by immuno-histochemically on the same region within the oocytes through their developmental stages. The distribution of type A and V are found on the animal or vegetal hemisphere only after the developmental stage IV according to the Normal Table of Nieuwkoop and Faber.Three dimensional studies of type A and V showed that they can be divided into type A_1 and A_2, type V_1 and V_2. These distributions are also confirmed by the western-blotting analysis of proteins extracted from the sections of animal and vegetal hemispher of mature eggs.Some of type A, V and P antibodies were used for the studies of their distribution on the blastula stage of fertilized eggs. Their localizations were exactly the same as in the matured egg.