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退化或功能障碍是多种疾病的病理生理学基础和/或促成因素，包括阿尔茨海默病、运动神经元疾病和家族性自主神经障碍，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研究这些疾病的实验方法的发展以及治疗策略的设计至关重要。我们专注于阐明调节胆碱能神经递质表型的机制，我们的数据表明，在小鼠中枢神经系统和大脑的原代培养中，特定的骨形态发生蛋白（BMPs）显著上调这种表型的表达。bmp在诱导和维持神经元胆碱能表型中的作用的细胞和分子机制仍然未知，这将是当前建议的主题。我们具体建议：1。鉴定BMPs介导胆碱能表型诱导的细胞内信号分子；描述胆碱能基因位点内对bmp有反应的启动子区域，并确定与bmp反应区域结合的蛋白质转录因子。这些研究将在野生型小鼠和具有候选转录因子基因靶向突变的动物的原代神经元培养物上进行，这些候选转录因子可能介导bmp对胆碱能表型的影响。研究将集中在Smad蛋白和其他转录调节因子的信号功能，报告基因构建中胆碱能特异性启动子的表达，以及候选转录因子的DNA结合。此外，我们提出了动物实验来验证体内给药bmp可以通过神经解剖学工具纠正中隔海马损伤引起的胆碱能功能下调的假设。总的来说，这项研究将为bmp在大脑中的作用机制提供基础和临床前数据，以及它们在治疗影响胆碱能神经元的神经系统疾病方面的潜力。

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.