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

这项研究着眼于神经系统内神经细胞复杂排列的形成机制。近年来，很明显，一种名为PTEN的特定因子调节着神经细胞正确组装起作用的神经系统所需的许多过程。除此之外，人们还意识到，抑制PTEN有助于神经细胞在变性条件下存活，也可以在神经系统损伤后恢复神经细胞的生长。因此，本研究建议研究神经系统中控制PTEN功能的机制。我们将研究(1)PTEN如何被神经细胞中的其他因素调控，(2)PTEN如何帮助神经细胞相互沟通，(3)不同蛋白质对PTEN的放松调控如何对神经系统的形成产生影响，以及(4)我们如何翻译这些信息，以便开发出以非常特定的方式抑制PTEN功能的新方法。我们将使用不同的实验，这些实验具有新颖性和互补性。首先，我们将使用干细胞，我们将其分化为神经元。我们将用已知可以改变PTEN功能的DNA序列来操纵这些细胞。通过这种方式，我们可以产生细胞的克隆，这些克隆可以被放大到几乎无限的数量。利用这些干细胞，我们将研究PTEN功能的变化如何影响神经细胞，以及哪些细胞机制参与了这些影响。其次，我们将研究PTEN在鸡胚胎发育过程中的功能。在这项研究中，我们将使用一套临时操作的雏鸟。我们将测试我们的干细胞产生的神经细胞是否整合到雏鸡的神经系统中，我们还将测试在雏鸡神经系统中直接操作PTEN是否会影响神经细胞之间的连接建立。这种方法帮助我们避免了在常用动物模型(如小鼠)中进行昂贵和耗时的实验。这项工作将帮助我们了解PTEN在神经系统中是如何调控的，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。

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.