Institute of Biomedicine and Molecular Immunology, National Research Council, via Ugo La Malfa, 153-90100, Palermo, Italy.
Dev Growth Differ. 2013 Feb;55(2):237-46. doi: 10.1111/dgd.12023. Epub 2013 Jan 10.
The sea urchin provides a relatively simple and tractable system for analyzing the early stages of embryo development. Here, we use the sea urchin species, Paracentrotus lividus, to investigate the role of Alix in key stages of embryogenesis, namely the egg fertilization and the first cleavage division. Alix is a multifunctional protein involved in different cellular processes including endocytic membrane trafficking, filamentous (F)-actin remodeling, and cytokinesis. Alix homologues have been identified in different metazoans; in these organisms, Alix is involved in oogenesis and in determination/differentiation events during embryo development. Herein, we describe the identification of the sea urchin homologue of Alix, PlAlix. The deduced amino acid sequence shows that Alix is highly conserved in sea urchins. Accordingly, we detect the PlAlix protein cross-reacting with monoclonal Alix antibodies in extracts from P. lividus, at different developmental stages. Focusing on the role of PlAlix during early embryogenesis we found that PlAlix is a maternal protein that is expressed at increasingly higher levels from fertilization to the 2-cell stage embryo. In sea urchin eggs, PlAlix localizes throughout the cytoplasm with a punctuated pattern and, soon after fertilization, accumulates in larger puncta in the cytosol, and in microvilli-like protrusions. Together our data show that PlAlix is structurally conserved from sea urchin to mammals and may open new lines of inquiry into the role of Alix during the early stages of embryo development.
海胆为分析胚胎发育早期阶段提供了一个相对简单且易于处理的系统。在这里,我们使用海胆物种 Paracentrotus lividus 来研究 Alix 在胚胎发生的关键阶段(即卵子受精和第一次卵裂)中的作用。Alix 是一种多功能蛋白,参与包括内吞膜运输、丝状(F)肌动蛋白重塑和胞质分裂在内的不同细胞过程。Alix 同源物已在不同的后生动物中被鉴定出来;在这些生物中,Alix 参与卵母细胞发生和胚胎发育过程中的决定/分化事件。在此,我们描述了 Alix 海胆同源物 PlAlix 的鉴定。推断的氨基酸序列表明 Alix 在海胆中高度保守。因此,我们在来自 P. lividus 的不同发育阶段的提取物中检测到与单克隆 Alix 抗体发生交叉反应的 PlAlix 蛋白。在关注 PlAlix 在早期胚胎发生中的作用时,我们发现 PlAlix 是一种母源性蛋白,从受精到 2 细胞胚胎阶段,其表达水平逐渐升高。在海胆卵中,PlAlix 分布在整个细胞质中,呈现出点状模式,受精后不久,它在细胞质中积累在更大的点状结构中,并在微绒毛样突起中积累。总之,我们的数据表明,PlAlix 从海胆到哺乳动物的结构保守,这可能为研究 Alix 在胚胎发育早期阶段的作用开辟新的研究途径。
Mar Biotechnol (NY). 2016-6