In vivo Profiling of Glial and Neuronal Activities in Psychostimulant Abuse

精神兴奋剂滥用中神经胶质和神经元活动的体内分析

• 批准号: 7851185
• 负责人: Philip K Liu
• 金额: $ 62.78万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-15 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7851185
• 关键词: Abstinence; Actins; Affect; Amphetamine Addiction; Amphetamines; Animals; Autopsy; Blood - brain barrier anatomy; Brain; Brain Diseases; Cell Culture Techniques; Cells; Cerebrum; Chronic; Chronic Disease; Contrast Media; Data; Detection; Development; Discrimination; Doctor of Philosophy; Dose; Drug Addiction; Drug Exposure; Drug abuse; Event; Exposure to; Eyedrops; FOS gene; Gene Activation; Gene Expression; Genes; Glial Fibrillary Acidic Protein; Goals; Imaging Techniques; Immunohistochemistry; In Situ; In Situ Hybridization; Individual; Intraperitoneal Injections; Knockout Mice; Life; Link; Longitudinal Studies; Magnetic Resonance Imaging; Maps; Mediating; Messenger RNA; Methods; Modification; Molecular; Molecular Profiling; Monitor; Mus; Neuroglia; Neurons; Neurosciences; Neurosciences Research; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Play; Polymerase Chain Reaction; Preparation; Productivity; Proteins; RNA Probes; Resolution; Reverse Transcription; Role; Route; Sampling; Sensitivity and Specificity; Signal Transduction; Slice; Societies; Staging; Staining method; Stains; Substance abuse problem; Techniques; Time; Toxin; Up-Regulation; Wild Type Mouse; addiction; base; behavioral sensitization; dosage; drug of abuse; imaging probe; in vivo; iron oxide; member; nanoparticle; novel; nutrition; protein expression; psychostimulant; public health relevance; receptor; response; segregation; spatiotemporal; stimulant abuse

DESCRIPTION (provided by applicant): In vivo profiling of glial and neuronal activities in psychostimulant abuse (PI: Christina H Liu, PhD) Drug addiction is a chronic brain disorder that severely hampers productivity of many members of our society. One of the most used drugs of abuse is amphetamine. Most neuroscience studies on amphetamine have focused on the morphological and molecular adaptations of neurons in response to chronic drug exposure. Electrophysiological data from cell cultures or brain slice preparations show that an important component of drug abuse is the interaction that occurs between glia and neurons during neuronal activations arising from repeat exposure to amphetamine. Although we know that glial cells are essential to the survival of neurons, providing critical support, nutrition, and toxin clearance to the neurons, our understanding of the role of glia in drug addiction is very limited. To overcome this limitation, we have developed a novel in vivo probe to visualize the changes in gene activities in neurons and glia, using magnetic resonance imaging (MRI) in live brains after exposure to amphetamine. We have demonstrated the utility of this method for discrimination of cells that express different but specific gene activities; i.e., neuronal-based elevation of cfos messenger RNA (mRNA) from glia-based GFAP mRNA. Because our detection technique does not require the use of postmortem samples, we are able to study the longitudinal profiles of different cerebral gene activities during and after exposure to drugs. Here, we aim to study gene activation during amphetamine sensitization in living C57Black6 mice, which involves repeated usage of, abstinence from, and re-exposure to the drug. We will utilize our novel technique that uses mRNA-targeted MR contrast probes to detect altered endogenous gene activities that result from amphetamine exposure in neurons and glia. The targets to be investigated in this project are mRNAs of glial fibrillary acidic protein (GFAP) and neuronal cFos protein. Our goals for this project are to use this novel MRI technique to enhance current understanding of the interactions between glial and neuronal cells at different stages of amphetamine exposure. We propose the following specific aims: (1) determine the optimal dosage and retention profiles of MRI probes in live brains; (2) determine the gene activity profiles of glial and neuronal cells in mouse brains in vivo after amphetamine, at different stages of exposure; and (3) explore the neuron-mediated glial response in the development of the amphetamine addiction phenotype. PUBLIC HEALTH RELEVANCE: In this project, we will utilize a novel Magnetic Resonance Imaging (MRI) technique that we have developed to detect altered gene expression in neurons and glia of live animal affected by addictive drugs. The goals of this project are to enhance current understanding of the interaction between glial and neuronal cells at different stages of amphetamine exposure. Such enhancement will bring a step closer to real-time analysis of neurophysiologic events that occur in patients with substance abuse and addiction.

描述（由申请人提供）：精神刺激虐待中神经胶质和神经元活动的体内分析（PI：Christina H Liu，PhD）药物成瘾是一种慢性脑疾病，严重妨碍了我们社会许多成员的生产力。最常用的滥用药物之一是苯丙胺。大多数关于苯丙胺的神经科学研究都集中在响应慢性药物暴露的神经元的形态和分子适应上。来自细胞培养物或脑切片制剂的电生理数据表明，药物滥用的重要组成部分是在反复暴露于苯丙胺引起的神经元激活期间神经元与神经元之间发生的相互作用。尽管我们知道神经胶质细胞对于神经元的生存至关重要，为神经元提供关键的支持，营养和毒素清除率，但我们对胶质成瘾中神经胶质作用的理解非常有限。为了克服这一局限性，我们开发了一种新颖的体内探针，以使用苯丙胺暴露后活体大脑中的磁共振成像（MRI）来可视化神经元和神经胶质的基因活性的变化。我们已经证明了这种方法对表达不同但特定基因活性的细胞歧视的实用性。即，基于神经胶质的GFAP mRNA的CFOS Messenger RNA（mRNA）的基于神经元的升高。由于我们的检测技术不需要使用后验尸样本，因此我们能够研究暴露于药物的不同脑基因活性的纵向谱。在这里，我们旨在研究活在C57BLACK6小鼠中苯丙胺致敏期间的基因激活，该小鼠涉及反复使用，禁欲和重新暴露对药物的使用。我们将利用使用靶向mRNA的MR对比探针的新技术来检测因神经元和神经胶质中苯丙胺暴露而导致的内源性基因活性的改变。在该项目中要研究的靶标是神经纤维纤维酸性蛋白（GFAP）和神经元CFOS蛋白的mRNA。我们的该项目的目标是使用这种新颖的MRI技术来增强对苯丙胺暴露不同阶段的神经胶质细胞和神经元细胞之间相互作用的当前理解。我们提出以下特定目的：（1）确定活大脑中MRI探针的最佳剂量和保留概况； （2）确定在暴露阶段，在苯丙胺后，体内小鼠大脑中神经胶质细胞和神经元细胞的基因活性谱； （3）在苯丙胺成瘾表型的发展中探索神经元介导的神经胶质反应。公共卫生相关性：在这个项目中，我们将利用一种新型的磁共振成像（MRI）技术来检测受成瘾性药物影响的活动物神经元和神经胶质的基因表达改变。该项目的目标是增强对苯丙胺暴露不同阶段的神经胶质细胞和神经元细胞之间相互作用的当前理解。这种增强将使对药物滥用和成瘾的患者发生的神经生理事件的实时分析更加接近实时分析。