GENE EXPRESSION AND CA2+ REGULATION IN BRAIN AGING AND ALZHEIMER'S DISEASE

脑衰老和阿尔茨海默病中的基因表达和 CA2 调节

• 批准号: 7475678
• 负责人: KUEY-CHU CHEN
• 金额: $ 29.36万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7475678
• 关键词: APP-PS1; Age; Age-Months; Aging; Alzheimer' s Disease; Animal Model; Animals; Biological Assay; Biological Markers; Cells; Cognition; Collaborations; Collection; Data; Detection; Gene Expression; Gene Proteins; Genes; Goals; Hippocampus (Brain); Image Analysis; In Situ Hybridization; Measures; Mediating; Messenger RNA; Microglia; Modeling; Mus; Neuroglia; Neurons; Pathway interactions; Phase; Physiological; Play; Polymerase Chain Reaction; Preparation; Process; Protein Overexpression; Proteins; Proteomics; Rattus; Regulation; Relative (related person); Risk; Risk Factors; Role; Signal Pathway; Signaling Molecule; Signaling Protein; Slice; Steroids; Techniques; Testing; Time; Viral; Work; age group; age related; aging brain; cell type; hippocampal pyramidal neuron; middle age; normal aging; novel; pathological aging; research study; steroid hormone; steroid hormone receptor; transcription factor

The overall goal of this project is to identify genes/proteins that play a causal role in brain aging/risk for AD. We will test the hypothesis that interactions of steroids with Ca2+ signaling pathways are underlying causal factors that act in early mid-life, using gene microarrays, combined with sophisticated biostatistical analyses and real-time PCR, protein assays and in situ hybridization. In the first three specific aims, we will extend our previous work by determining which changes in gene expression take place earliest in normal aging rats and also in an animal model of AD, the APP/PS1 mouse. Comparison of these two models will reveal how normal aging transitions into pathological aging and will identify risk factors for AD. In addition, we will carry out microarray analyses of single neurons and gila to that test the hypothesis that aging is a process of neuronal involution and gUallmicroglial activation. We will coordinate this project closely with Project 3, in order to determine the temporal relationship between altered gene expression and Ca 2+ dysregulation assessed by physiological techniques. In the last two specific aims, we will test the hypothesis that steroid hormones and their coordinating transcription factors and Ca2+-regulated signaling molecules play a role in inducing changes in gene expression that mimic those in aging. These studies will be carried out using viral-mediated overexpression/knockdown of steroid hormone receptors or Ca2+-regulated signaling proteins in mixed cultures of hippocampal cells. Subsequent experiments will be carried out in cell-type specific cultures (neurons, gila, microglia) in order to identify where the changes in gene expression originate. Gene expression changes elicited by these treatments will be determined first using microarrays and subsequently, real-time PCR of selected panels of aging biomarkers. Taken together, these studies will elucidate the role of both factors previously implicated in brain aging (steroid hormones, Ca 2+- regulating molecules) and will identify novel factors and pathways, as well. Further, this Project in collaboration with Project 3 and Cores A, B and C will define more clearly the relationship of gene expression cascades to Ca 2+ dysregulation in aging/AD.

该项目的总体目标是确定在AD的大脑衰老/风险中起因果作用的基因/蛋白质。我们将检验以下假设：类固醇与Ca2+信号通路的相互作用是使用基因微阵列在中年早期起作用的基本因果因素，并结合了复杂的生物统计学分析和实时PCR，蛋白质分析和原位杂交。在前三个特定目标中，我们将通过确定最早在正常衰老大鼠以及AP APP/PS1小鼠的动物模型中确定基因表达的哪些变化来扩展以前的工作。对这两个模型的比较将揭示正常的衰老过渡到病理衰老的方式，并确定AD的危险因素。此外，我们还将对单个神经元和吉拉进行微阵列分析，以检验衰老是神经元相关和guallmicroglial激活过程的假设。我们将与项目3紧密协调该项目，以确定基因表达改变的时间关系和通过生理技术评估的Ca 2+失调之间的时间关系。在最后两个 具体目的，我们将检验以下假设：类固醇激素及其配位转录因子和Ca2+调节的信号传导分子在诱导模仿衰老的基因表达的变化中起作用。这些研究将使用病毒介导 在海马细胞混合培养物中，类固醇激素受体或Ca2+调节的信号蛋白的过表达/敲低。随后的实验将在细胞型特异性培养物（神经元，吉拉，小胶质细胞）中进行，以确定基因表达的变化起源。这些治疗引起的基因表达变化将首先使用微阵列，然后是随后的衰老生物标志物的实时PCR确定。综上所述，这些研究将阐明以前与脑衰老（类固醇激素，Ca 2+调节分子）有关的两个因素的作用，并将确定新的因素和途径。此外，该项目与项目3和核心A，B和C合作将更清楚地定义基因表达级联反应与衰老/AD中Ca 2+失调的关系。