Tang P, Sharpe C R, Mohun T J, Wylie C C
Department of Anatomy, St. George's Hospital Medical School, London, UK.
Development. 1988 Jun;103(2):279-87. doi: 10.1242/dev.103.2.279.
Immunocytochemical studies using a monoclonal anti-porcine vimentin antibody reveal a well-organized pattern of staining in Xenopus laevis oocytes, eggs and early embryos. The positions of Xenopus vimentin and desmin in two-dimensional (2D) polyacrylamide gels were first established by immunoblotting of muscle Triton extracts with anti-intermediate filament antibodies (anti-IFA), which cross-react with all intermediate filament proteins (IFPs). The anti-porcine vimentin reacts with vimentin and desmin in muscle 2D immunoblots, but only reacts with one polypeptide in oocyte blots in the position predicted for vimentin (Mr 55 x 10(3), pI 5.6). Using an anti-sense probe derived from a Xenopus vimentin genomic clone in RNase protection assays, we show that expression of vimentin begins in previtellogenic oocytes. The level of expression remains constant throughout oogenesis and in unfertilized eggs. These data suggest that vimentin is expressed in oocytes and eggs. Most interestingly, the immunocytochemical results also show that vimentin is present in the germ plasma of oocytes, eggs and early embryos. It is therefore possible that vimentin has an important role in the formation or behaviour of early germ line cells.
使用单克隆抗猪波形蛋白抗体进行的免疫细胞化学研究显示,非洲爪蟾的卵母细胞、卵和早期胚胎中有组织良好的染色模式。通过用抗中间丝抗体(抗IFA)对肌肉Triton提取物进行免疫印迹,首先确定了非洲爪蟾波形蛋白和结蛋白在二维(2D)聚丙烯酰胺凝胶中的位置,该抗体与所有中间丝蛋白(IFP)发生交叉反应。抗猪波形蛋白在肌肉二维免疫印迹中与波形蛋白和结蛋白发生反应,但在卵母细胞印迹中仅与预测波形蛋白位置的一种多肽发生反应(Mr 55 x 10(3),pI 5.6)。在核糖核酸酶保护试验中,使用源自非洲爪蟾波形蛋白基因组克隆的反义探针,我们发现波形蛋白的表达始于卵黄生成前的卵母细胞。在整个卵子发生过程和未受精卵中,表达水平保持恒定。这些数据表明波形蛋白在卵母细胞和卵中表达。最有趣的是,免疫细胞化学结果还表明波形蛋白存在于卵母细胞、卵和早期胚胎的生殖质中。因此,波形蛋白在早期生殖系细胞的形成或行为中可能具有重要作用。