Neurons Generated from APP/APLP1/APLP2 Triple Knockout Embryonic Stem Cells Behave Normally in Vitro and in Vivo: Lack of Evidence for a Cell Autonomous Role of the Amyloid Precursor Protein in Neuronal Differentiation
Neurons Generated from APP/APLP1/APLP2 Triple Knockout Embryonic Stem Cells Behave Normally in Vitro and in Vivo: Lack of Evidence for a Cell Autonomous Role of the Amyloid Precursor Protein in Neuronal Differentiation
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DOI：
10.1002/stem.296
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发表时间：
2010-03-31
期刊：
影响因子：
5.2
通讯作者：
De Strooper, Bart
中科院分区：
文献类型：
--
作者：
Bergmans, Bruno A.;Shariati, S. Ali M.;De Strooper, Bart
阿尔茨海默病淀粉样前体蛋白 (APP) 与许多神经生物学过程有关，但支持证据仍然是间接的。研究因两个部分冗余的 APP 同源物 APLP1 和 APLP2 的存在而令人困惑。 APP/APLP1/APLP2 三重敲除 (APP tKO) 小鼠表现出鹅卵石无脑畸形，并且在围产期是致命的。为了解决这个问题，我们生成了 APP 三重敲除胚胎干 (ES) 细胞，并在体外和体内将其分化为 APP 三重敲除神经元。与野生型（WT）ES细胞来源的神经元相比，APP tKO神经元形成同样纯的神经元培养物，具有未改变的体外迁移能力，具有相似的极性获得，并且能够延伸长神经突并形成活跃的兴奋性突触。这些数据在带有 APP tKO 神经元的嵌合小鼠体内得到证实，这些神经元表达 WT 背景大脑中存在的增强型绿色荧光蛋白 (eGFP)。结果表明，APP 蛋白家族的丢失对这些关键的神经元过程没有重大影响，并且 APP 所涉及的明显多种功能可能以分子冗余为特征。我们的干细胞培养提供了一个极好的工具来规避 APP/APLP 三重敲除小鼠缺乏活力的问题，并将有助于在体外和体内进一步探索这种有趣蛋白质的功能。干细胞2010； 28：399-406
Alzheimer's disease amyloid precursor protein (APP) has been implicated in many neurobiologic processes, but supporting evidence remains indirect. Studies are confounded by the existence of two partially redundant APP homologues, APLP1 and APLP2. APP/APLP1/APLP2 triple knockout (APP tKO) mice display cobblestone lissencephaly and are perinatally lethal. To circumvent this problem, we generated APP triple knockout embryonic stem (ES) cells and differentiated these to APP triple knockout neurons in vitro and in vivo. In comparison with wild-type (WT) ES cell-derived neurons, APP tKO neurons formed equally pure neuronal cultures, had unaltered in vitro migratory capacities, had a similar acquisition of polarity, and were capable of extending long neurites and forming active excitatory synapses. These data were confirmed in vivo in chimeric mice with APP tKO neurons expressing the enhanced green fluorescent protein (eGFP) present in a WT background brain. The results suggest that the loss of the APP family of proteins has no major effect on these critical neuronal processes and that the apparent multitude of functions in which APP has been implicated might be characterized by molecular redundancy. Our stem cell culture provides an excellent tool to circumvent the problem of lack of viability of APP/APLP triple knockout mice and will help to explore the function of this intriguing protein further in vitro and in vivo. STEM CELLS 2010; 28: 399-406