BMP actions on cholinergic cells

描述（由申请人提供）：中枢和外周胆碱能神经元的变性或功能障碍是多种疾病的病理生理学基础和/或促成多种疾病的病理生理学，包括阿尔茨海默病、运动神经元疾病和家族性自主神经功能障碍，以及可能与衰老相关的记忆丧失。了解胆碱能表型及其调节机制对于研究这些疾病的实验方法的发展和治疗策略的设计至关重要。我们已经集中在阐明机制，调节胆碱能神经递质表型和我们的数据显示，特定的骨形态发生蛋白（BMP）显着上调这种表型在小鼠中枢神经系统的原代培养物的表达，并在体内的大脑。BMP诱导和维持神经元胆碱能表型的细胞和分子作用机制仍不清楚，将是本提案的主题。具体来说，我们建议：1。鉴定介导骨形成蛋白诱导胆碱能表型的细胞内信号分子。描绘胆碱能基因位点内对BMP反应的启动子区域，并鉴定与这些BMP反应区域结合的蛋白质转录因子。将对从野生型小鼠和具有编码候选转录因子的基因靶向突变的动物获得的原代神经元培养物进行研究，所述候选转录因子可能介导BMP对胆碱能表型的影响。调查将集中在Smad蛋白和其他转录调节因子的信号传导功能，胆碱能特异性启动子在报告构建体中的表达，以及候选转录因子的DNA结合。此外，我们提出动物研究，以测试的假设，即在体内施用骨形态发生蛋白可以纠正隔-海马病变诱发的胆碱能功能的下调，使用神经解剖工具。总之，这项研究将提供有关BMP在大脑中的作用机制及其用于治疗影响胆碱能神经元的神经系统疾病的潜力的基础和临床前数据。

DESCRIPTION (provided by applicant): Degeneration or malfunction of central and peripheral cholinergic neurons underlies and/or contributes to the pathophysiology of a variety of disorders including Alzheimer's Disease, Motor Neuron Disorders, and Familial Dysautonomias, and possibly memory loss associated with aging. Understanding the cholinergic phenotype and the mechanisms regulating it is crucial to the development of experimental approaches to study these diseases, and to the design of treatment strategies. We have focused on the elucidation of mechanisms that regulate the cholinergic neurotransmitter phenotype and our data show that specific bone morphogenetic proteins (BMPs) dramatically upregulate the expression of this phenotype in primary cultures of mouse central nervous system, and in the brain in vivo. The cellular and molecular mechanisms of action of BMPs in inducing and maintaining the neuronal cholinergic phenotype remain unknown and will be the subject of the current proposal. Specifically we propose: 1. To identify the intracellular signaling molecules that mediate the induction of the cholinergic phenotype by BMPs, and 2. To delineate the promoter regions within the cholinergic gene locus that respond to BMPs and to identify protein transcription factors that bind to those BMP-responsive regions. The studies will be performed on primary neuronal cultures obtained from wild type mice and from animals with targeted mutations of genes encoding candidate transcription factors that may mediate the effects of BMPs on the cholinergic phenotype. Investigations will focus on the signaling functions of Smad proteins and other transcription regulators, expression of cholinergic-specific promoters in reporter constructs, and DNA binding of candidate transcription factors. In addition, we propose animal studies to test the hypothesis that administration of BMPs in vivo can correct septo-hippocampal lesion evoked downregulation of cholinergic function using neuroanatomical tools. Taken together this research will provide basic and preclinical data on the mechanisms of action of BMPs in the brain and the potential for their use in treating neurological disorders affecting cholinergic neurons.