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非洲爪蟾卵母细胞和早期胚胎中波形蛋白及新型波形蛋白相关蛋白的鉴定

Identification of vimentin and novel vimentin-related proteins in Xenopus oocytes and early embryos.

作者信息

Torpey N P, Heasman J, Wylie C C

机构信息

Department of Zoology, University of Cambridge, UK.

出版信息

Development. 1990 Dec;110(4):1185-95. doi: 10.1242/dev.110.4.1185.

Abstract

We have made antibodies against fusion proteins of Xenopus vimentin. We show for the first time the distribution of vimentin in larval stages, where it is found in cells of mesenchymal origin, and in radial glial cells. In sections of Xenopus oocytes and early embryos, immunocytochemistry reveals the presence of an extensive cytoplasmic network, distributed in an animal-vegetal gradient. Germ plasm stains particularly strongly. The form of the IF proteins in this network is unusual. In immunoblot experiments the anti-vimentin antibodies detect a number of distinct proteins. We have identified those that are the products of the two known vimentin genes, by injection of synthetic mRNA transcribed from cloned vimentin cDNAs into oocytes, followed by two-dimensional Western blotting. This has demonstrated unambiguously that one Xenopus vimentin, Vim1, is present in oocytes and early embryos. However, two other immunoreactive proteins detected in Triton extracts of oocytes and early embryos are not the products of Vim1, since depletion of vimentin mRNA by antisense oligonucleotide injection has no effect on the synthesis of these proteins. These results suggest that novel IF-like proteins are expressed in Xenopus oocytes and early embryos.

摘要

我们制备了针对非洲爪蟾波形蛋白融合蛋白的抗体。我们首次展示了波形蛋白在幼虫阶段的分布情况,它存在于间充质起源的细胞以及放射状胶质细胞中。在非洲爪蟾卵母细胞和早期胚胎的切片中,免疫细胞化学显示存在广泛的细胞质网络,呈动植物梯度分布。生殖质染色特别强烈。该网络中中间丝蛋白的形式不同寻常。在免疫印迹实验中,抗波形蛋白抗体检测到一些不同的蛋白质。通过将从克隆的波形蛋白cDNA转录的合成mRNA注射到卵母细胞中,然后进行二维蛋白质印迹,我们鉴定出了那些是两个已知波形蛋白基因产物的蛋白质。这明确证明了一种非洲爪蟾波形蛋白Vim1存在于卵母细胞和早期胚胎中。然而,在卵母细胞和早期胚胎的Triton提取物中检测到的另外两种免疫反应性蛋白不是Vim1的产物,因为通过反义寡核苷酸注射耗尽波形蛋白mRNA对这些蛋白质的合成没有影响。这些结果表明,非洲爪蟾卵母细胞和早期胚胎中表达了新型的类中间丝蛋白。

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