TARGET GENE IDENTIFICATION OF EMX TRANSCRIPTION FACTORS

拟议研究的长期目标是了解 大脑皮层发育的分子和细胞基础。 我们要研究的具体假设是，胚胎 大脑皮层限制性转录因子Emx1和/或Emx2 用表达限制调控靶基因的转录 划分发育中的大脑皮层。采取的方法是 检测不同组织中Emx1和Emx2的内源性表达水平 小鼠神经细胞系，包括几个来自皮质的细胞系， 使用Northern blotts。选择内源性EMX较少的细胞系 表达，可诱导的Emx1和/或Emx2基因将稳定 整合使用磷酸钙转染法。EMX 在去除低水平的基因后，基因将高度表达 从细胞培养上清液中提取四环素。上调的mRNA转录本- 或在Einx诱导后下调，将使用RDA和 聚合酶链式反应差异显示。将对减法产品进行筛查 胚胎大脑皮层的原位杂交表达 其中最有趣的将是DNA测序。 理解导致正常的分子相互作用 大脑皮层的发育可以揭示出 表征了这个器官的问题，从阅读困难到 精神分裂症。

The long term objectives of the research proposed is to understand the molecular and cellular basis for the development of the cerebral cortex. The specific hypothesis we aim to investigate is that the embryonic cerebral cortex-restricted transcription factors Emx1 and/or Emx2 regulate the transcription of target genes with limits of expression demarcating the developing cerebral cortex. The approach taken is to determine endogenous expression levels of Emx1 and Emx2 in various murine neural cell lines, including several derived from the cortex, using Northern blots. Choosing a cell line with little endogenous Emx expression, an inducible Emx1 and/or Emx2 gene will be stably integrated using a calcium phosphate transfection protocol. The Emx genes will be highly expressed after removal of low levels of tetracycline from the cell culture medium. mRNA transcripts that are up- or downregulated after Einx induction will be identified using RDA and PCR differential display. Subtraction products will be screened for embryonic cerebral cortex expression using in situ hybridization, and the most interesting of these will be characterized by DNA sequencing. Understanding the molecular interactions responsible for normal development of the cerebral cortex could shed light on the multitude of characterized problems with this organ, ranging from dyslexia to schizophrenia.