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

描述（由申请人提供）：神经系统的适当发育依赖于在适当的时间和地点发生的许多细胞过程。神经系统发育所需要的一个细胞过程是某些mRNA转录物的适当定位和翻译。这些转录本的定位依赖于mRNA结合蛋白，这是一类负责运输的分子，在某些情况下，负责mRNA货物的翻译。Zipcode结合蛋白1 （ZBP1）是一种mRNA结合蛋白，负责转运和调节-肌动蛋白mRNA的翻译。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基因的功能对正常发育具有重要意义。目前的研究有两个主要目标：1)研究ZBP1在轴突生长调控和引导中的作用；2)研究ZBP1在-actin mRNA的转运和翻译中的作用。为了确定ZBP1在这些过程中的作用，将采用慢病毒载体介导的shRNA干扰ZBP1，以及ZBP1敲除小鼠模型。本建议还在原代神经元培养中使用创新的荧光蛋白和报告蛋白，以实现上述目标。研究-actin mRNA的定位和翻译是至关重要的，因为它编码一种结构蛋白，这种结构蛋白是生长锥运动和引导所必需的，因此是建立神经系统正确连接所必需的。该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转录物的局部翻译；这一信息将进一步加深我们对神经系统整体发育的认识。神经系统的发育是一个过程，在这个过程中，神经元必须找到通往合适目标的路，并与之建立联系。如果这种情况不正常发生，可能会导致许多疾病状态。本研究探讨了在发育过程中负责mRNA运输的蛋白ZBP1的作用；这种蛋白质的缺失可能导致神经系统发育不正常，从而导致神经系统疾病。

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.