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;这种极性对于正常功能是必不可少的。细胞极性通过许多机制建立，包括特定信使RNA（mRNA）的调节性亚细胞定位。局部mRNA导致局部蛋白质合成和使用;这对于许多细胞的正常发育和神经元的突触可塑性极其重要。这项研究的目的是加深我们对所有细胞，特别是神经元中mRNA定位的基本过程的理解。 该项目的第一个目标是调整体内技术，用于一个特定的模式生物，线虫秀丽隐杆线虫。C.出于各种原因，elegans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，所以个别标记的神经元可以很容易地在体内观察到。它有一个完整的测序和充分表征的基因组。最后，制造转基因动物和快速分离突变体很容易。该项目的第一部分将是产生转基因C。携带荧光标记的mRNA的线虫，这些mRNA不均匀地分布在单个识别的神经元中。在这些转基因产生后，它们将用于诱变和筛选以分离在神经元中具有异常mRNA定位的突变体。使用C中可用的工具。在线虫中，突变体的分离将很容易导致参与这一过程的一些基因的鉴定。将克隆两个或更多的这些基因，并将在该资助所涵盖的两年期结束前开始对其表达和蛋白质产物进行初步表征。这些基因的信息和mRNA定位在C。elegans在体内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.