Transcriptome Analysis of Identified Cells in Developing Xenopus laevis CNS

描述(申请人提供)：非洲爪哇一直是研究神经发育的有价值的模型系统。最近几年，它的实用性已经被其他模型系统黯然失色，仅仅是因为它缺乏分子和细胞操作所需的遗传工具。然而，许多特性使非洲爪哇成为研究、操纵和测试体内神经元发育范式的独特和不可替代的模型。建议的研究重点有三个方面：1)为非洲爪哇提供急需的基因转录本和microRNAs的序列和表达的遗传信息，这些基因转录本和microRNAs在非洲爪哇脑的几个区域，在几种已鉴定的神经细胞类型和几个发育阶段，2)产生试剂，用于分子/遗传操作已确定的转录本和信号通路，包括反义Morolino结构，siRNA和目标基因的细胞类型特异性表达，以及3)使用这些试剂来探索候选调控基因在完整动物神经发生中的功能。拟议的工作将产生丰富的试剂，这些试剂将显着加强非洲爪哇作为生物医学研究的实验模型的使用，并允许进行跨物种比较研究，这将丰富其他实验系统的研究。

DESCRIPTION (provided by applicant): Xenopus laevis has been a valuable model system for studying neural development. In more recent years its utility has been eclipsed by other model systems simply because it lacks the genetic tools required for molecular and cellular manipulations. Nevertheless, many features make Xenopus a unique and irreplaceable model for studying, manipulating and testing neuronal developmental paradigms in vivo. The focus of the proposed studies is threefold: 1) to provide the Xenopus community with much needed genetic information on the sequence and expression of gene transcripts and microRNAs in several regions of the X. laevis brain, in several identified neural cell types and at several developmental stages, 2) to generate reagents for molecular/genetic manipulations of identified transcripts and signaling pathways, including antisense morpholino constructs, siRNA and cell-type specific expression of genes of interest and 3) to use the reagents to probe the function of candidate regulatory genes in neurogenesis in the intact animal. The proposed work will generate a wealth of reagents that will significantly enhance use of Xenopus as an experimental model for biomedical research and permit cross-species comparative studies that will enrich research in other experimental systems.