ZBP1-mediated transport and local translation of B-actin mRNA in growth cones

描述(由申请人提供)：神经系统的正常发育依赖于在适当的时间和地点发生的一些细胞过程。神经系统正常发育所需的一个这样的细胞过程就是某些mRNA转录本的适当定位和翻译。这些转录本的定位依赖于mRNA结合蛋白，这是一类负责运输和在某些情况下翻译它们的mRNA货物的分子。邮编结合蛋白1(ZBP1)是一种信使核糖核酸结合蛋白，负责肌动蛋白mRNA的转运和翻译。肌动蛋白对神经系统的发育很重要，因为它是构成生长锥的主要结构蛋白之一，生长锥是发育中的神经元的寻路结构。目前这项提案的目的是确定ZBP1的缺失如何影响神经系统发育，特别是生长锥。研究ZBP1的功能具有重要的意义，因为它对适度的总体发展至关重要。目前的建议有两个主要目的：1)研究ZBP1在轴突生长调节和引导中的作用；2)研究ZBP1在肌动蛋白mRNA的运输和翻译中的作用。为了确定ZBP1在这些过程中的作用，我们将采用慢病毒载体介导的对ZBP1的shRNA干扰，以及ZBP1基因敲除小鼠的模型。为了达到上述目的，该提案还在原代神经元培养中使用了创新的荧光蛋白和报告。研究肌动蛋白mRNA的定位和翻译是至关重要的，因为它编码一种结构蛋白，这种结构蛋白是正确的生长锥运动和引导所必需的，因此是建立正确的神经系统连接所必需的。这项建议的长期目标是了解轴突生长和生长锥引导所需的基本机制，例如生长锥内mRNA转录本的局部翻译；这些信息将进一步加深我们对神经系统作为一个整体的适当发育的了解。神经系统的发育是一个过程，在这个过程中，神经元必须找到自己的方式，并与适当的目标建立联系。如果这不适当地发生，可能会导致一些疾病状态。这项建议研究了ZBP1蛋白的作用，该蛋白负责在发育过程中运输mRNA；该蛋白的丢失可能导致神经系统的不适当发育，从而导致神经疾病状态。

DESCRIPTION (provided by applicant): The proper development of the nervous system is dependent on a number of cellular processes occurring at the appropriate time and location. One such cellular process that is required for proper nervous system development is the appropriate localization and translation of certain mRNA transcripts. The localization of these transcripts is dependent on mRNA binding proteins, a class of molecules responsible for the transport, and in some cases, translation of their mRNA cargo. Zipcode binding protein 1 (ZBP1) is an mRNA binding protein that is responsible for the transport and regulates the translation of ¿-actin mRNA. ¿-actin is important for the development of the nervous system because it is one of the major structural proteins that composes the growth cone, the pathfinding structure of a developing neuron. The aim of the current proposal is to determine how the loss of ZBP1 affects nervous system development and specifically, the growth cone. It is of great importance to study the function of ZBP1 because it is essential for appropriate gross development. The current proposal has two main goals: 1) to examine the role of ZBP1 in the regulation of axon outgrowth and guidance and 2) to examine the role of ZBP1 in the transport and translation of ¿-actin mRNA. To determine the role of ZBP1 in these processes, lentiviral vector-mediated shRNA interference to ZBP1 will be employed, as well as a ZBP1 knockout mouse model. This proposal also uses innovative fluorescent proteins and reporters in primary neuronal cultures in order to achieve the above stated aims. It is essential to study the localization and translation of ¿-actin mRNA because it encodes a structural protein that is required for proper growth cone motility and guidance, and therefore is required for the correct connectivity of the nervous system to be established. The long-term objective of this proposal is to gain an understanding of the basic mechanisms that are required for axon outgrowth and growth cone guidance, such as local translation of mRNA transcripts within the growth cone; this information will further our knowledge about the appropriate development of the nervous system as a whole. The development of the nervous system is a process during which neurons must find their way to and make connections with their appropriate targets. If this does not happen properly, a number of disease states may result. This proposal investigates the role of the protein ZBP1, which is responsible for transporting mRNA during development; the loss of this protein may result in inappropriate development of the nervous system and thus, a neurological disease state.