PACAP REGULATION OF NEUROGENESIS AND SURVIVAL

PACAP 对神经发生和生存的调节

• 批准号: 2037735
• 负责人: Emanuel Murray DiCicco-Bloom
• 金额: $ 19.05万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2037735
• 关键词: DNA footprinting; adenylate cyclase; autocrine; autoradiography; cell cycle; cyclic AMP; enzyme activity; gene induction /repression; genetic regulatory element; growth factor receptors; immunocytochemistry; laboratory rat; lipid metabolism; neurogenesis; neurotrophic factors; northern blottings; nucleotide metabolism; phosphatidylinositols; pituitary hormones; protein tyrosine kinase; receptor expression; tissue /cell culture; transcription factor; vasoactive intestinal peptide

While the growth of neuronal population has been well-characterized in vivo, underlying molecular mechanisms remains undefined. The goal of this proposal is to define the role of PACAP (pituitary adenylate cyclase activating peptide) in regulating the three principal mechanisms contributing to neuronal population growth including 1) stimulating mitosis of precursor or neuroblasts, 2) enhancing survival of proliferating precursors and 3) promoting longer-term survival through action of target-derived neurotrophic factor s. PACAP may be critical to normal brain development, since this neuropeptide influences neurogenic and survival of multiple prenatal and postnatal as well as peripheral and central populations. Our specific aim are to define the effects of PACAP on neuroblast mitosis and survival as well as expression of the trk family mRNA's. Second, characterized the effect of PACAP on second messenger systems: cAMP and phosphatidyl inositol (PI) metabolism. Third, define dht regulation and expression of PACAP protein and mRNA in the neurogenetic populations, and characterized peptide autocrine and autoregulatory activity. Fourth, characterized PACAP induction of DNA binding activities that interact with cAMP response elements (CRE's) and other DNA sites in target neuronal gene promoter. Our strategy employs pure populations of mitotic neuroblasts from embryonic sympathetic and sensory ganglia, cerebral cortical neuroepithelium and postnatal cerebellar cortex to identity population- specific and age-dependent PACAP regulation. We examine mitogenic and trophic effects by assaying cell number, [3H]thymidine incorporation, and neurite outgrowth. cAMP and PACAP protein will be assessed y radioimmunoassay an PI metabolism by ion-exchange chromatography. mRNA;s will be assayed by Norther analysis. PACAP induction of population- specific binding activities will be characterized by gel retardation, DNAse I footprinting, and UV crosslinking procedures. By characterizing PACAP actions in peripheral and central neurogenetic populations, we many define mechanisms by which the single peptide, PACAP, differentially regulates population-specific neurogenesis and survival. Hopefully we may learn how extracellular signals, acting via second messenger and transcriptional pathways, alter neuroblast division, survival and trophic factor receptor expression to generate diverse neuronal populations. Insight into second messenger and transcription pathways underlying PACAP mitogenic and trophic activity may identify loci where disease processes intervene to produce dysraphic sates, holoprosencephaly, and systems disorders.

虽然神经元种群的增长已经充分表现 体内，潜在的分子机制仍然不确定。 目标 该建议是定义PACAP的作用（垂体腺苷酸环化酶 激活肽）调节三种主要机制 促进神经元种群的增长，包括1）刺激 前体或神经细胞有丝分裂，2）增强的生存率 增殖前体和3）通过 目标衍生的神经营养因子的作用。 PACAP可能很关键 由于这种神经肽会影响正常的大脑发育 多个产前和产后以及的神经源性和存活 外围和中央人口。 我们的具体目的是定义PACAP对神经细胞有丝分裂的影响 以及TRK家族mRNA的生存以及表达。 第二， 表征了PACAP对第二通信系统的影响：营地和 磷脂酰肌醇（PI）代谢。 第三，定义DHT调节和 PACAP蛋白和mRNA在神经遗传种群中的表达，以及 肽自分泌和自动调节活动的表征。 第四， 表征PACAP诱导的DNA结合活性的相互作用 具有cAMP响应元素（CRE）和目标中的其他DNA位点 神经元基因启动子。 我们的策略采用了纯有丝分裂神经细胞的种群 胚胎交感神经和感觉神经节，脑皮质 神经上皮和产后小脑皮质与身份种群 - 特定和年龄依赖的PACAP调节。 我们检查有丝分裂和 通过分析细胞数，[3H]胸苷掺入和 神经突生长。将评估营地和PACAP蛋白 放射免疫分子通过离子交换色谱法代谢。 mRNA; s 将通过Norther分析来测定。 PACAP诱导人口 - 特定的结合活动将以凝胶延迟为特征， DNase I足迹和UV交联过程。 通过表征外周和中央神经遗传学中的PACAP动作 种群，我们许多定义了单个肽的机制， PACAP，差异调节人群特异性神经发生和 生存。希望我们可以学习细胞外信号，通过 第二信使和转录途径，改变神经细胞部门， 生存和营养因子受体表达以产生多样化 神经元种群。 深入了解第二信使和转录 PACAP有丝分裂和营养活性的途径可能会鉴定 疾病处理干预以产生异性疾病的基因座， 全脑脑和系统疾病。