Quantitative proteomics of Xenopus oocytes, eggs, blastomeres, and nuclear transplants

 描述（由申请人提供）： 抽象的。该研究小组最近发表了第一个发育胚胎的定量蛋白质组学分析（Sci Rep. 2014; 4：4365）。PMID：24626130）。我们研究了单个非洲爪蟾胚胎在六个阶段的发展。我们最初量化了至少四个发育阶段中4,000种蛋白质的表达变化;使用改进的基因组数据库重新分析数据，将我们的鉴定扩展了50%。数据已在Scien-tific Reports网站、Xenbase和PeptideAtlas上提供给社区。 我们将通过定量监测从单个非洲爪蟾卵母细胞、卵和卵裂球中分离的蛋白质的表达和磷酸化状态来扩展我们的初步研究。还将从垂直于发育轴的冷冻切片测量定量蛋白质表达变化，以鉴定沿着所选细胞谱系、来自该谱系的外植体和核移植后的卵母细胞中的图案化梯度。这些研究的结果将是一个前所未有的资源爪蟾社区提供详细的数据，在发展过程中的蛋白质表达的演变，在胚胎和核移植后的染色质重组。我们将与Xenbase、PeptideAtlas和VectorBase合作，尽可能广泛地传播我们的结果。

 DESCRIPTION (provided by applicant): Abstract. This research team recently published the first quantitative proteomic analysis of a devel- oping embryo (Sci Rep. 2014; 4: 4365. PMID: 24626130). We studied single Xenopus laevis embryos at six stages of development. We initially quantified the expression change of 4,000 proteins across at least four stages of development; reanalysis of the data with an improved genomic database has extended our identifications by 50%. Data have been made available to the community on the Scien- tific Reports web site, at Xenbase, and at PeptideAtlas. We will extend our initial study by quantitatively monitoring the expression and phosphorylation status of proteins isolated from single Xenopus oocytes, eggs, and blastomeres. Quantitative protein expression changes will also be measured from cryosections perpendicular to developmental axes to identify patterning gradients, along a selected cell lineage, from an explant of that lineage, and in oocytes following nuclear transplantation. The results of these studies will be an unprecedented resource to the Xenopus community by providing detailed data on the evolution of protein expression during development, and chromatin reorganization in embryos and following nuclear transplantation. We will work with Xenbase, PeptideAtlas, and VectorBase to disseminate our results as widely as possible.