BMP actions on cholinergic cells

描述(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基础和/或促成各种疾病的病理生理学，包括阿尔茨海默病、运动神经元障碍、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于发展研究这些疾病的实验方法和设计治疗策略至关重要。我们致力于阐明调节胆碱能神经递质表型的机制，我们的数据显示，在原代培养的小鼠中枢神经系统中，特定的骨形态发生蛋白(BMPs)显著上调这一表型的表达，并在体内的脑中表达。BMPs诱导和维持神经元胆碱能表型的细胞和分子作用机制尚不清楚，将是当前提议的主题。具体地说，我们建议：1.确定介导BMPs诱导胆碱能表型的细胞内信号分子；2.确定胆碱能基因座内对BMPs反应的启动子区域，并确定与这些BMP反应区域结合的蛋白转录因子。这些研究将在从野生型小鼠和动物获得的原代神经元培养上进行，这些神经元编码候选转录因子的基因具有靶向性突变，这些基因可能介导BMPs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，报告结构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，我们建议进行动物实验，以验证体内注射BMPs可以纠正隔区-海马区损害引起的胆碱能功能下调的假说。综上所述，这项研究将提供关于BMPs在大脑中的作用机制以及它们在治疗影响胆碱能神经元的神经疾病方面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.