Watanabe Masanao, Itoh Keiji, Abe Kuniya, Akizawa Toshifumi, Ikenishi Kohji, Furusawa Mitsuru
Furusawa MorphoGene Project, Exploratory Research for Advanced Technology (ERATO), Research Development Corporation of Japan (JRDC), Tsukuba 300-26, Japan.
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Setsunan University, Hirakata 573-01, Japan.
Dev Growth Differ. 1992 Apr;34(2):223-231. doi: 10.1111/j.1440-169X.1992.tb00011.x.
In order to investigate whether a vasa-like protein is present in germ line cells of Xenopus, antibodies were produced which react specifically with synthetic oligopeptides of sequences from near the N- or C-termini or with one including the DEAD box of the Drosophila vasa protein. Only the antibody against the oligopeptide including the DEAD box reacted strongly with germ plasm (GP) or with cytoplasm of germ line cells of Xenopus embryos by immunofluorescence microscopy. By immunoelectron microscopy, the antibody was demonstrated to react with the GP-specific structure, germinal granules, in cleaving embryos, and with their derivatives in the germ line cells of embryos at stages extending from gastrula to feeding tadpole. It also reacted with mitochondria not only in the GP and the germ line cells but also in somatic cells, and with myofibrils in muscle cells. By Western blotting, the antibody was shown to react with several bands of Mr 42-69 ± 10 in protein samples from Xenopus embryos. In samples from Drosophila ovaries, it reacted with a Mr 71 ± 10 band which was probably the vasa protein. This indicates the possibility that Xenopus embryos contain several DEAD family proteins. One of these is present on germinal granules, resembling the vasa protein on polar granules of Drosophila.
为了研究非洲爪蟾生殖系细胞中是否存在类瓦萨蛋白，制备了能与来自果蝇瓦萨蛋白N端或C端附近序列的合成寡肽或与包含DEAD框的寡肽特异性反应的抗体。通过免疫荧光显微镜观察，只有针对包含DEAD框的寡肽的抗体与非洲爪蟾胚胎生殖质（GP）或生殖系细胞的细胞质发生强烈反应。通过免疫电子显微镜观察，该抗体在分裂胚胎中与GP特异性结构、生殖颗粒发生反应，并在从原肠胚到摄食蝌蚪阶段的胚胎生殖系细胞中与它们的衍生物发生反应。它不仅在GP和生殖系细胞中，而且在体细胞中与线粒体发生反应，还与肌肉细胞中的肌原纤维发生反应。通过蛋白质印迹法，该抗体在非洲爪蟾胚胎的蛋白质样品中与几条分子量为42 - 69±10的条带发生反应。在果蝇卵巢的样品中，它与一条分子量为71±10的条带发生反应，该条带可能是瓦萨蛋白。这表明非洲爪蟾胚胎可能含有几种DEAD家族蛋白。其中一种存在于生殖颗粒上，类似于果蝇极颗粒上的瓦萨蛋白。
