Approaches to study protein complexes and signaling systems during neural circuit formation using ES cell-derived neurons.

使用 ES 细胞衍生的神经元研究神经回路形成过程中的蛋白质复合物和信号系统的方法。

• 批准号: BB/I022392/1
• 负责人: Britta Eickholt
• 金额: $ 89.31万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI022392%2F1
• 关键词: Approaches; study; protein; complexes; signaling

This research looks at the mechanisms involved in the formation of the complex arrangement of nerve cells within the nervous system. In recent years, it became apparent that a specific factor called PTEN regulates many processes required for nerve cells to proper assemble a functioning nervous system. In addition to this, it was realised that inhibition of PTEN helps the survival of nerve cells under conditions of degeneration, and, also, that it can re-install growth of nerve cells following injury to the nervous system. Therefore, this research proposes to investigate the mechanisms that control PTEN function in the nervous system. We will study (1) how PTEN is regulated by other factors in the nerve cell, (2) how PTEN helps nerve cells to communicate with each other, (3) how deregulation of PTEN by different proteins has an effect on the formation of the nervous system, and (4) how we can translate this information in order to develop novel ways in which we can inhibit the function of PTEN in a very specific way. We will use different experiments, which have novel and complementary advantages. Firstly, we will use stem cells, which we will differentiate into neurons. We will manipulate these cells with DNA sequences known to alter PTEN functions. In this way we generate clones of cells that can be amplified to virtual unlimited quantities. Using these stem cells, we will investigate how alteration in PTEN function affects the nerve cells and which cellular mechanisms are involved in mediating these effects. Secondly, we will study PTEN function in chick embryos as they develop. In this research we will use a set of temporary manipulations of the chick. We will test if our nerve cells produced from stem cells integrate into the nervous system of the chick, and we will also test if direct manipulation of PTEN in the nervous system of the chick affects the establishment of connection between nerve cells. This approach helps us to avoid expensive and time-consuming experiments in commonly used animal models such as the mouse. This work will help us understand how PTEN is regulated in the nervous system, and will help us to develop future treatment strategies that help neurons to re-integrate into neuronal networks following injury or during degenerative conditions.

这项研究着眼于神经系统内神经细胞复杂排列形成的机制。近年来，很明显，一种称为PTEN的特定因子调节神经细胞正确组装功能神经系统所需的许多过程。除此之外，人们意识到抑制PTEN有助于神经细胞在变性条件下的存活，并且还可以在神经系统损伤后重新安装神经细胞的生长。因此，本研究拟探讨神经系统中控制PTEN功能的机制。我们将研究（1）PTEN如何受到神经细胞中其他因素的调节，（2）PTEN如何帮助神经细胞相互交流，（3）不同蛋白质对PTEN的失调如何影响神经系统的形成，以及（4）我们如何翻译这些信息，以开发新的方法，我们可以以非常特定的方式抑制PTEN的功能。我们将使用不同的实验，这些实验具有新颖和互补的优势。首先，我们将使用干细胞，我们将分化成神经元。我们将用已知改变PTEN功能的DNA序列操纵这些细胞。通过这种方式，我们产生了可以扩增到几乎无限数量的细胞克隆。利用这些干细胞，我们将研究PTEN功能的改变如何影响神经细胞，以及哪些细胞机制参与介导这些效应。其次，我们将研究鸡胚发育过程中PTEN的功能。在这项研究中，我们将使用一套临时操纵小鸡。我们将测试由干细胞产生的神经细胞是否整合到小鸡的神经系统中，我们还将测试在小鸡神经系统中直接操纵PTEN是否会影响神经细胞之间连接的建立。这种方法有助于我们避免在常用的动物模型（如小鼠）中进行昂贵且耗时的实验。这项工作将帮助我们了解PTEN如何在神经系统中调节，并将帮助我们开发未来的治疗策略，帮助神经元在损伤后或退行性疾病期间重新整合到神经元网络中。

项目成果

Phosphorylation of the actin binding protein Drebrin at S647 is regulated by neuronal activity and PTEN.

• DOI: 10.1371/journal.pone.0071957
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Kreis P;Hendricusdottir R;Kay L;Papageorgiou IE;van Diepen M;Mack T;Ryves J;Harwood A;Leslie NR;Kann O;Parsons M;Eickholt BJ
• 通讯作者: Eickholt BJ

Capillary Isoelectric Focusing of Akt Isoforms Identifies Highly Dynamic Phosphorylation in Neuronal Cells and Brain Tissue.

• DOI: 10.1074/jbc.m115.700138
• 发表时间: 2016-05-06
• 期刊: The Journal of biological chemistry
• 作者: Schrötter S;Leondaritis G;Eickholt BJ
• 通讯作者: Eickholt BJ