In Vivo Two-Photon Imaging of Cortical Activity in Alcohol-Dependent Mice

酒精依赖小鼠皮质活动的体内双光子成像

基本信息

批准号：
8635067
负责人：
JOHN J. WOODWARD
金额：
$ 21.49万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-15 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8635067
关键词：
Affect Alcohol abuse Alcohol consumption Alcohol dependence Alcoholism Alcohols Behavior Blood Vessels Blood flow Brain Brain imaging Breeding Calcium Caliber Cells Cerebrum Chronic Cognition Cognitive Complication Coupling Cues Dependence Diffusion Dyes Economic Burden Electroencephalography Ethanol Ethanol dependence Experimental Models Future Glutamates Goals Heavy Drinking Image Impairment In Vitro Individual Interneurons Intervention Label Laser Scanning Microscopy Lead Light Maps Measures Medial Methods Modeling Molecular Monitor Morbidity - disease rate Mouse Strains Mus Neurobiology Neuroglia Neurons Population Prefrontal Cortex Preparation Process Property Proteins Relapse Reliance Reporting Research Design Resolution Rest Sensory Sensory Process Signal Transduction Slice Specificity Stimulus System Techniques Testing Thinking Tissue Sample Tissues Toxic effect Visible Radiation Visual Visual Cortex Withdrawal alcohol cue alcohol effect alcohol research arteriole base cell type cerebrovascular drinking fluorophore hippocampal pyramidal neuron in vivo light microscopy mortality mouse model new technology novel novel strategies prevent problem drinker public health relevance response selective expression sensor treatment strategy two-photon vapor visual stimulus

项目摘要

The goal of this project is to use in vivo two-photon laser scanning microscopy (2PLSM) to examine changes in neuronal activity in ethanol-dependent mice. Chronic drinking that leads to dependence is associated with marked changes in brain function including impairments in cortical processing and cognition. These impairments likely underlie the transition to heavy drinking and elucidating the mechanisms that underlie these changes is critical towards developing treatments and interventions that restore control over behavior. A key limitation in the study of the mechanistic actions on alcohol on brain function is the reliance upon in vitro techniques to monitor neuronal function. These approaches, while extremely valuable, are hindered by the loss of relevant sensory input and disruption of normal circuitry during preparation of the in vitro tissue sample. While brain imaging approaches can avoid this complication, they are compromised by a lack of specificity with regard to spatial resolution and neuronal sub-type. In this application, we will use in vivo 2PLSM to image neuronal activity and neurovascular function in alcohol-dependent mice. In Aim 1, we will utilize a recently developed line of mice that, when crossed with a specific Cre-driver mouse line, expresses the calcium-sensor protein GCaMP3 in a defined sub-population of neurons. Neuronal calcium dynamics will be monitored in glutamatergic pyramidal neurons and fast-spiking interneurons by crossing the GCaMP3 mice with the Wfs1-Tg2-CreERT2 line and Pvalb-2A-Cre line; respectively. Activity will be monitored both during rest and during presentation of visual cues that reliably induce activity in visual cortex neurons. In Aim 2, neurovascular coupling (e.g., activity-dependent changes in local brain blood flow) will be monitored by the use of a dye recently shown by the Co-I to effectively signal changes in arteriolar blood flow. Together, results from these two complementary aims will establish the technique of in vivo 2PLSM in the study of alcohol action and will support future studies designed to analyze the effects of alcohol on cortical function.

该项目的目标是使用体内双光子激光扫描显微镜（2PLSM）检查乙醇依赖小鼠神经元活动的变化。长期饮酒导致依赖性的原因与大脑功能的显着变化有关，包括皮质处理和认知障碍。这些损害可能是向酗酒的转变并阐明这些变化背后的机制对于开发恢复控制的治疗和干预措施至关重要行为。酒精对大脑作用机制研究的一个关键局限性功能是依赖体外技术来监测神经元功能。这些方法虽然非常有价值，但会因相关感官输入的丢失而受到阻碍以及体外组织样本制备过程中正常电路的破坏。尽管脑成像方法可以避免这种并发症，但它们因缺乏关于空间分辨率和神经元亚型的特异性。在这个应用程序中，我们将使用体内 2PLSM 对神经元活动和神经血管功能进行成像酒精依赖小鼠。在目标 1 中，我们将利用最近开发的小鼠品系，当与特定的 Cre-driver 小鼠品系杂交时，表达钙传感器特定神经元亚群中的 GCaMP3 蛋白。神经元钙动力学将在谷氨酸能锥体神经元和快速尖峰中间神经元中进行监测将 GCaMP3 小鼠与 Wfs1-Tg2-CreERT2 系和 Pvalb-2A-Cre 系杂交；分别。休息期间和视觉呈现期间的活动都将受到监控可靠地诱导视觉皮层神经元活动的线索。在目标 2 中，神经血管耦合（例如，局部脑血流的活动依赖性变化）将通过以下方式进行监测 Co-I 最近证明使用一种染料可以有效地发出小动脉变化信号血流（量。这两个互补目标的结果将共同建立体内 2PLSM 技术在酒精作用研究中的应用并将支持未来的研究旨在分析酒精对皮质功能的影响。