mRNA localization in C.elegans neurons in vivo

描述(由申请人提供)： 大多数真核细胞是不对称的或极性的；这种极性是正常功能所必需的。细胞极性是通过许多机制建立的，包括特定信使RNA(MRNAs)的调节亚细胞定位。定位的mRNA导致局部蛋白质的合成和使用；这对许多细胞的正常发育和神经元的突触可塑性非常重要。这项研究的目的是加深我们对所有细胞，特别是神经元中信使核糖核酸定位的基本过程的理解。 该项目的第一个目标是使体内技术用于一种特定的模式生物--线虫秀丽线虫。由于各种原因，线虫非常适合于研究神经元中的基本细胞过程。这种动物是半透明的，有一个非常简单和描述良好的身体计划，所以个体标记的神经元很容易在体内观察到。它有一个完全测序和良好特征的基因组。最后，制造转基因动物和快速分离突变体是很容易的。该项目的第一部分将是产生转基因线虫，这种线虫携带荧光标记的mRNAs，这些mRNAs不均匀地分布在单个已识别的神经元中。在这些转基因物质产生后，它们将被用于突变和筛选，以分离在神经元中具有异常mRNA定位的突变体。利用线虫可用的工具，分离突变株将很容易导致识别这一过程中涉及的一些基因。这些基因中的两个或更多将被克隆，其表达和蛋白质产品的初步特征将在这笔赠款所涵盖的两年期结束之前开始。这些基因的信息以及线虫体内mRNA定位的信息将增加我们对导致细胞极性和功能的基本过程的理解。

DESCRIPTION (provided by applicant): Most eukaryotic cells are asymmetric or polar; this polarity is essential for normal function. Cellular polarity is established via a number of mechanisms, including the regulated subcellular localization of specific messenger RNAs (mRNAs). Localized mRNA leads to local protein synthesis and use; this is extremely important for normal development of many cells and for synaptic plasticity in neurons. The goal of this research is to deepen our understanding of the basic processes underlying mRNA localization in all cells and, particularly, in neurons. The first aims of the project are to adapt an in vivo technique for use in a particular model organism, the nematode Caenorhabditis elegans. C. elegans is ideally suited for studies of basic cellular processes in neurons for a variety of reasons. The animal is translucent and has a very simple and well described body plan, so individual marked neurons can readily be observed in vivo. It has a completely sequenced and well characterized genome. Finally, it is easy to make transgenic animals and to rapidly isolate mutants. The first part of the project will be to generate transgenic C. elegans that carry fluorescently tagged mRNAs that are non-uniformly distributed within individual identified neurons. After these transgenics have been generated, they will be used in mutageneses and screens to isolate mutants that have abnormal mRNA localization in neurons. Using the tools available in C. elegans, the isolation of mutants will readily lead to the identification of some of the genes involved in this process. Two or more of these genes will be cloned and initial characterization of their expression and protein products will begin before the end of the two year period covered by this grant. The information on these genes and on mRNA localization in C. elegans in vivo will increase our understanding of basic processes that lead to cell polarity and function.