Chronic CeA CRF overexpression on gene expression in PVN CRF-expressing cells

慢性 CeA CRF 过表达对 PVN CRF 表达细胞中基因表达的影响

• 批准号: 7331587
• 负责人: Elizabeth Irene Flandreau
• 金额: $ 2.73万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7331587
• 关键词: Acute; Address; Adrenal Glands; Adult; Amygdaloid structure; Anhedonia; Animals; Antidepressive Agents; Behavior; Behavioral; Brain; Brain-Derived Neurotrophic Factor; Cells; Chronic; Chronic stress; Condition; Corticotropin; Corticotropin-Releasing Hormone; Depressed mood; Disruption; Elevation; Endocrine; Event; Feedback; Fluorescence-Activated Cell Sorting; Functional disorder; Gene Expression; Glucocorticoid Receptor; Glucocorticoids; Green Fluorescent Proteins; Human; Hyperactive behavior; Hypothalamic structure; Lentivirus Vector; Literature; Major Depressive Disorder; Measures; Mediating; Mental Depression; Messenger RNA; Methods; Monitor; Mouse Strains; Mus; Neurons; Oocytes; Output; Patients; Pattern; Pituitary Gland; Play; Polymerase Chain Reaction; Prevalence; Production; Protein Overexpression; Public Health; Research; Role; Signal Transduction; Source; Stress; Structure; Subfamily lentivirinae; Swimming; Symptoms; Techniques; Technology; Testing; Thinking; Time; Transgenic Mice; Transgenic Organisms; Vasopressins; Withdrawal; base; depressive symptoms; design; dexamethasone suppression test; hypothalamic-pituitary-adrenal axis; mRNA Expression; mature animal; molecular pathology; novel; preference; promoter; recombinase; vector

DESCRIPTION (provided by applicant): Project Summary: The hypothalamic pituitary adrenal (HPA) axis is hyperactive in many patients with major depressive disorder (MOD). This dysfunction normalizes with successful treatment and may be a commonality behind treatments. Novel treatments should target the source of HPA axis hyperactivity: presumably dysregulated corticotropin releasing factor (CRF) from the paraventricular hypothalamus (PVN). PVN activity is regulated by the coordination of numerous limbic structures and plasticity in one or more of these regions may cause enhanced PVN CRF output and subsequent HPA axis dysfunction. One likely culprit is the central amygdala (CeA), another region expressing high concentrations of CRF and thought to mediate stress- induced behavior. The Aims of the present research are to determine (1) if, in addition to behavioral effects, chronic increased CRF drive from the CeA reproduces endocrine changes associated with depressive symptoms and (2) if chronically overexpressing CRF in the CeA results in gene expression changes in PVN CRF neurons that correlate with the observed endocrine and behavioral disruptions. A lentiviral vector will overexpress CRF in CRF-expressing neurons (LVCRFp3.OCRF) in the CeA of adult animals. CRF-expressing cells will be easily identifiable by creating a transgenic mouse in which CRF- expressing cells also express green fluorescent protein (GFP). These cells can be isolated via fluorescence activated cell sorting and subjected to gene expression analysis to compare the CRF and non CRF- expressing cells in the PVN and CeA under basal conditions or after chronically overexpressing CRF in the CeA. It is expected that chronic CeA CRF overexpression will result in HPA axis hyperactivity that can be explained at least partially by gene expression changes in PVN CRF cells. Relevance to Public Health: Chronic elevations in CRF may initiate a chain of events altering gene expression and endocrine signaling and eventually leading to symptoms of depression. CRF is released by numerous brain structures but the relationship between CRF in these regions is incompletely understood because previous technology has been unable to mimic chronic CRF elevations. Using superior techniques, CRF will be overexpressed in one region and then CRF cells in another region will be isolated to identify gene-expression changes that may correlate with observed endocrine and behavioral alterations.

描述（由申请人提供）：项目概述：下丘脑垂体肾上腺（HPA）轴在许多重度抑郁症（MOD）患者中是过度活跃的。这种功能障碍随着成功的治疗而正常化，可能是治疗背后的共性。新的治疗应该针对HPA轴过度活跃的来源：可能是室旁下丘脑（PVN）的促肾上腺皮质激素释放因子（CRF）失调。PVN的活动是由许多边缘结构的协调和可塑性在一个或多个这些地区可能会导致增强PVN CRF输出和随后的HPA轴功能障碍。一个可能的罪魁祸首是中央杏仁核（CeA），另一个表达高浓度CRF的区域，被认为是介导压力诱导行为的区域。本研究的目的是确定（1）除了行为效应之外，来自CeA的慢性增加的CRF驱动是否再现与抑郁症状相关的内分泌变化，以及（2）CeA中慢性过表达CRF是否导致PVN CRF神经元中的基因表达变化，这些变化与观察到的内分泌和行为干扰相关。慢病毒载体将在成年动物的CeA中的CRF表达神经元（LVCRFp3.OCRF）中过表达CRF。通过产生转基因小鼠，表达CRF的细胞也表达绿色荧光蛋白（GFP），可以容易地鉴定表达CRF的细胞。这些细胞可以通过荧光激活细胞分选分离，并进行基因表达分析，以比较在基础条件下或在CeA中长期过表达CRF后PVN和CeA中的CRF和非CRF表达细胞。预计慢性CeA CRF过表达将导致HPA轴过度活跃，这至少可以部分地通过PVN CRF细胞中的基因表达变化来解释。与公共卫生的相关性：慢性CRF升高可能引发一系列事件，改变基因表达和内分泌信号，最终导致抑郁症状。CRF由许多脑结构释放，但这些区域中CRF之间的关系尚未完全了解，因为以前的技术无法模拟慢性CRF升高。使用上级技术，CRF将在一个区域中过表达，然后将另一个区域中的CRF细胞分离以鉴定可能与观察到的内分泌和行为改变相关的基因表达变化。