BDNF PROMOTER ELEMENTS IN THE DEVELOPING HYPOTHALAMUS

描述（改编自申请人的描述）：本项目的目标 建议是双重的。 第一是要描述的监管和作用， 脑源性神经营养因子（BDNF）在下丘脑。 二是 主要研究者作为独立研究者的发展。 脑源性神经营养因子（BDNF）属于神经生长因子（NGF）家族的神经营养因子（NTs） 其功能对于正常的神经系统发育至关重要。更 最近，人们赞赏它在正常运作中发挥关键作用， 成年人的大脑。 调查人员最近描述了一个 有趣的NT缺陷小鼠表型， 小鼠变得肥胖或过度活跃。 在转录水平上， 表型的特征在于BDNF在细胞内的差异调节。 下丘脑他们推测，表型的差异可能是由于 BDNF的转录控制。BDNF有四种启动子， 每一个都与它们自己的非翻译外显子沿着， 外显子。 本提案将评估这些外显子中每一个的表达和功能 特别是下丘脑核的方法 模型 通过研究每个未翻译的BDNF外显子的表达， 研究人员可以确定哪些启动子元件是关键的和特异的 到特定的下丘脑核团。 在具体目标1中，他们将分析 这些非翻译外显子中的每一个以细胞特异性方式表达 在下丘脑中。他们将通过利用lacZ报告器来实现这一点 对每个外显子具有特异性的构建体和随后转染这些 构建到器官型下丘脑培养物中。具体目标2将解决 最感兴趣的启动子的体内发育表达， 通过建立稳定转染的转基因小鼠系来实现特定目的1。 最后，特异性目标3将解决最特异性的 通过将突变引入小鼠胚胎干细胞， 研究人员已经证明， 转录。研究人员提出的小鼠只会 在需要靶向启动子元件的区域缺乏BDNF， 转录发生。通过这种方式，他们可以在分子水平上 靶向启动子的功能。

DESCRIPTION (Adapted from applicant's description): The goals of this proposal are twofold. The first is to characterize the regulation and role of brain derived neurotrophic factor (BDNF) in the hypothalamus. The second is the development of the principal investigator as an independent researcher. BDNF belongs to the nerve growth factor (NGF) family of neurotrophins (NTs) and its function is critical for normal nervous system development. More recently, it has been appreciated to play key roles in the proper functioning of the adult brain. The investigators have recently characterized an interesting phenotype of NT deficient mice whereby haplo-insufficient BDNF mice become obese or hyperactive. At the transcriptional level these phenotypes are characterized by differential regulation of BDNF within the hypothalamus. They hypothesize that divergence in phenotype may be due to the transcriptional control of BDNF. There are four promoters for BDNF that are each associated with their own untranslated exon along with a fifth coding exon. This proposal will evaluate the expression and function of each of these exons in particular hypothalamic nuclei by the use of both in vitro and in vivo models. By studying the expression of each untranslated BDNF exon, the investigators can determine which promoter elements are critical and specific to particular hypothalamic nuclei. In Specific Aim 1 they will analyze the expression of each of these untranslated exons in a cell specific manner within the hypothalamus. They will do this by utilizing lacZ reporter constructs specific for each exon and subsequent transfection of these constructs into organotypic hypothalamic cultures. Specific Aim 2 will address the in vivo developmental expression of the most interesting promoters from Specific Aim 1 by establishing a line of stably transfected transgenic mice. Finally, Specific Aim 3 will address the in vivo function of the most specific of these promoters by introducing a mutation into mouse embryonic stem cells of key promoter elements which the investigators have shown disrupt gene transcription. The mice that the investigators propose to generate will only be deficient in BDNF in areas that need the targeted promoter elements for transcription to occur. In this manner they can determine at a molecular level the function of the targeted promoter.