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非洲爪蟾Xpat蛋白是生殖质的主要成分,可能在其组织和定位中发挥作用。

Xenopus Xpat protein is a major component of germ plasm and may function in its organisation and positioning.

作者信息

Machado Rachel J, Moore Wendy, Hames Richard, Houliston Evelyn, Chang Patrick, King Mary Lou, Woodland Hugh R

机构信息

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.

出版信息

Dev Biol. 2005 Nov 15;287(2):289-300. doi: 10.1016/j.ydbio.2005.08.044. Epub 2005 Oct 7.

Abstract

In many animals, including Drosophila, C. elegans, zebrafish and Xenopus, the germ line is specified by maternal determinants localised in a distinct cytoplasmic structure called the germ plasm. This is consists of dense granules, mitochondria, and specific localised RNAs. We have characterised the expression and properties of the protein encoded by Xpat, an RNA localised to the germ plasm of Xenopus. Immunofluorescence and immunoblotting showed that this novel protein is itself a major constituent of germ plasm throughout oogenesis and early development, although it is also present in other regions of oocytes and embryos, including their nuclei. We found that an Xpat-GFP fusion protein can localise correctly in cultured oocytes, in early oocytes to the 'mitochondrial cloud', from which germ plasm originates, and in later oocytes to the vegetal cortex. The localisation process was microtubule-dependent, while cortical anchoring required microfilaments. Xpat-GFP expressed in late stage oocytes assembled into circular fields of multi-particulate structures resembling endogenous fields of germ plasm islands. Furthermore these structures could be induced to form at ectopic sites by manipulation of culture conditions. Ectopic Xpat-GFP islands were able to recruit mitochondria, a major germ plasm component. These data suggest that Xpat protein has an important role in Xenopus germ plasm formation, positioning and maintenance.

摘要

在许多动物中,包括果蝇、秀丽隐杆线虫、斑马鱼和非洲爪蟾,生殖系由定位于一种名为生殖质的独特细胞质结构中的母体决定因子所指定。生殖质由致密颗粒、线粒体和特定的定位RNA组成。我们已经对Xpat编码的蛋白质的表达和特性进行了表征,Xpat是一种定位于非洲爪蟾生殖质的RNA。免疫荧光和免疫印迹显示,这种新蛋白质本身是整个卵子发生和早期发育过程中生殖质的主要成分,尽管它也存在于卵母细胞和胚胎的其他区域,包括它们的细胞核。我们发现,Xpat-GFP融合蛋白能够在培养的卵母细胞中正确定位,在早期卵母细胞中定位于生殖质起源的“线粒体云”,在后期卵母细胞中定位于植物性皮质。定位过程依赖于微管,而皮质锚定则需要微丝。在晚期卵母细胞中表达的Xpat-GFP组装成多颗粒结构的圆形区域,类似于生殖质岛的内源性区域。此外,通过操纵培养条件,可以在异位位点诱导这些结构形成。异位的Xpat-GFP岛能够募集线粒体,线粒体是生殖质的主要成分。这些数据表明,Xpat蛋白在非洲爪蟾生殖质的形成、定位和维持中具有重要作用。

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