Doss Michael Xavier, Wagh Vilas, Schulz Herbert, Kull Meelis, Kolde Raivo, Pfannkuche Kurt, Nolden Tobias, Himmelbauer Heinz, Vilo Jaak, Hescheler Jürgen, Sachinidis Agapios
Center of Physiology and Pathophysiology, Institute of Neurophysiology, and Center of Molecular Medicine, University of Cologne (CMMC), Robert-Koch Str. 39, 50931 Cologne, GermanyMax-Delbrueck-Center for Molecular Medicine - MDC, Robert-Rössle Str. 10, 13092 Berlin, GermanyInstitute of Computer Science, University of Tartu, Liivi 2, 50409 Tartu, Estonia and Quretec Ltd, Ulikooli 6a, Tartu, EstoniaDepartment of Vertebrate Genomics, Max-Planck-Institute for Molecular Genetics, Ihnestr.73, D-14195 Berlin, Germany.
Genes Cells. 2010 Mar;15(3):209-28. doi: 10.1111/j.1365-2443.2010.01390.x. Epub 2010 Feb 24.
Brachyury(+) mesodermal cell population with purity over 79% was obtained from differentiating brachyury embryonic stem cells (ESC) generated with brachyury promoter driven enhanced green fluorescent protein and puromycin-N-acetyltransferase. A comprehensive transcriptomic analysis of brachyury(+) cells enriched with puromycin application from 6-day-old embryoid bodies (EBs), 6-day-old control EBs and undifferentiated ESCs led to identification of 1573 uniquely up-regulated and 1549 uniquely down-regulated transcripts in brachyury(+) cells. Furthermore, transcripts up-regulated in brachyury(+) cells have overrepresented the Gene Ontology annotations (cell differentiation, blood vessel morphogenesis, striated muscle development, placenta development and cell motility) and Kyoto Encyclopedia of Genes and Genomes pathway annotations (mitogen-activated protein kinase signaling and transforming growth factor beta signaling). Transcripts representing Larp2 and Ankrd34b are notably up-regulated in brachyury(+) cells. Knockdown of Larp2 resulted in a significantly down-regulation BMP-2 expression, and knockdown of Ankrd34b resulted in alteration of NF-H, PPARγ and PECAM1 expression. The elucidation of transcriptomic signatures of ESCs-derived brachyury(+) cells will contribute toward defining the genetic and cellular identities of presumptive mesodermal cells. Furthermore, there is a possible involvement of Larp2 in the regulation of the late mesodermal marker BMP-2. Ankrd34b might be a positive regulator of neurogenesis and a negative regulator of adipogenesis.
通过用短尾相关蛋白启动子驱动的增强型绿色荧光蛋白和嘌呤霉素 - N - 乙酰转移酶从分化的短尾胚胎干细胞（ESC）中获得了纯度超过79%的短尾相关蛋白（+）中胚层细胞群体。对来自6日龄胚状体（EBs）、6日龄对照EBs和未分化ESC的用嘌呤霉素处理富集的短尾相关蛋白（+）细胞进行全面的转录组分析，结果在短尾相关蛋白（+）细胞中鉴定出1573个独特上调和1549个独特下调的转录本。此外，在短尾相关蛋白（+）细胞中上调的转录本在基因本体注释（细胞分化、血管形态发生、横纹肌发育、胎盘发育和细胞运动）和京都基因与基因组百科全书通路注释（丝裂原活化蛋白激酶信号传导和转化生长因子β信号传导）方面有过度富集。代表Larp2和Ankrd34b的转录本在短尾相关蛋白（+）细胞中显著上调。敲低Larp2导致骨形态发生蛋白 - 2（BMP - 2）表达显著下调，敲低Ankrd34b导致神经丝蛋白 - H（NF - H）、过氧化物酶体增殖物激活受体γ（PPARγ）和血小板内皮细胞黏附分子1（PECAM1）表达改变。对ESC来源的短尾相关蛋白（+）细胞转录组特征的阐明将有助于确定推定中胚层细胞的遗传和细胞特性。此外，Larp2可能参与晚期中胚层标志物BMP - 2的调控。Ankrd34b可能是神经发生的正调节因子和脂肪生成的负调节因子。
