Activity Based Tagging of Neurons

描述(申请人提供)：大脑中的信息编码被认为反映在兴奋性神经元对给定刺激的反应模式中。这表明，从本质上讲，神经细胞类型是由各种刺激和条件定义的 招募它的电活动。大脑特定区域活动的改变与各种神经和精神疾病有关，治疗这些疾病的药物干预会改变特定回路的活动。复杂认知功能的细胞和分子变化，如学习和记忆，很可能发生在相关刺激激活的回路中的关键特定点。神经科学方面的大量努力是 然而，集中于定义这些激活的电路，当前可用的技术仅限于离散的大脑区域，缺乏细胞特异性，或者仅在单个时间点提供活动记录，从而防止从噪声或噪声中识别一致的网络激活模式 识别网络随时间变化以响应干预。我们将在这项资助中开发的方法使用一个单一的荧光标记来识别两个独立时间点的神经活动模式。这提供了比现有技术更多的优势，包括使用高通量自动成像技术分析大脑的能力，基于电生理、形态或分子研究的WALE动物的激活模式识别脑片中的特定细胞群的能力，以及将FACS分选技术应用于分离单个核以进行表观遗传学研究的能力。报告活动的两个时间点可以至少相隔一周，从而可以分析对长期行为或药物干预做出反应的回路变化和靶细胞群。这在确定这些疗法对大脑的关键变化方面应该是有用的。

DESCRIPTION (provided by applicant): Information encoding in the brain is thought to be reflected in the pattern of activation of excitatory neurons in response to a given stimulus. This suggests that, in essence, a neural cell type is defined by the various stimuli and conditions that recruit its electrical activity. Alterations in activity in specific brain regions are associated wth a variety of neurological and psychiatric diseases and the pharmacological interventions to treat these diseases alter activity in specific circuits. The cellular and molecular changes that underli complex cognitive functions such as learning and memory are likely to occur at critical specific points in the circuits activated by the relevant stimuli. A great deal of effort in neuroscience is focused on defining these activated circuits however, currently available techniques are limited to discrete brain areas, lack cellular specificity, or provide a record of activity at only a singl time point preventing the identification of consistent patterns of network activation from noise or the identification of network changes over time in response to intervention. The approach that we will develop in this grant uses a single florescent marker to identify neural activity patterns t two independent time points. This provides a number of advantages over existing technology including, the ability to analyze the brain using high throughput automated imaging techniques, to identify specific cell populations in brain slices based on their activation patterns in the whoe animal for electrophysiological, morphological, or molecular studies, and the ability to apply FACS sorting techniques to the isolation of individual nuclei for epigenetic studies. The two time points at which activity is reported can be separated by at least one week allowing the analysis of circuit changes and target cell populations that are responsive to prolonged behavioral or pharmacological intervention. This should be useful in identifying the critical changes in the brain in response to these therapies.