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

非洲爪蟾早期发育形态发生功能蛋白的研究。

基本信息

批准号：
03640605
负责人：
AMANO Minoru
金额：
$ 1.22万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03640605/
关键词：
Oocyte Body-axis Oocyte protein Oogenesis Monoclonal-antibody Xenopus laevis モノクロ-ナル抗体卵形成

项目摘要

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.

制备了抗早期卵母细胞胞质中不含卵黄蛋白的抗原的单克隆抗体。用105，000 × G成熟卵上清液作抗原制备了单克隆抗体。对卵巢组织切片进行免疫染色，筛选特异性抗体。用不同发育阶段的卵母细胞进行染色试验，获得了10株发育阶段特异性单克隆抗体。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，将其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。P型和N型抗体通过免疫组织化学在卵母细胞发育阶段的同一区域检测。根据Nieuwkoop和Faber的正态分布表，A型和V型在动物或植物半球的分布仅在发育阶段IV以后才有，对A型和V型的三维研究表明，它们可分为A_1和A_2型，V_1和V_2型。这些分布也被从成熟卵的动物和植物半球的切片中提取的蛋白质的western-blotting分析所证实，并用A、V和P型抗体研究了它们在受精卵囊胚期的分布。它们的定位与成熟卵中的定位完全相同。