Transcriptome Analysis of Identified Cells in Developing Xenopus laevis CNS

描述（由申请人提供）：非洲爪蟾是研究神经发育的有价值的模型系统。近年来，它的效用已黯然失色其他模型系统，只是因为它缺乏所需的分子和细胞操纵的遗传工具。然而，许多功能使非洲爪蟾一个独特的和不可替代的模型，用于研究，操纵和测试体内神经元发育范式。拟开展的研究的重点有三个方面：1）为非洲爪蟾群落提供急需的遗传信息，即X染色体几个区域中基因转录本和microRNA的序列和表达。在几种鉴定的神经细胞类型和几个发育阶段，2）产生用于鉴定的转录物和信号传导途径的分子/遗传操作的试剂，包括反义吗啉代构建体、siRNA和感兴趣基因的细胞类型特异性表达，和3）使用所述试剂来探测完整动物中神经发生中候选调控基因的功能。拟议的工作将产生丰富的试剂，将显着提高使用非洲爪蟾作为生物医学研究的实验模型，并允许跨物种的比较研究，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.