Activity Based Tagging of Neurons

描述（由申请人提供）：大脑中的信息编码被认为反映在兴奋性神经元响应给定刺激的激活模式中。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， 恢复它的电活动。特定脑区活动的改变与各种神经和精神疾病相关，治疗这些疾病的药物干预会改变特定回路的活动。作为学习和记忆等复杂认知功能基础的细胞和分子变化可能发生在相关刺激激活的回路中的关键特定点。在神经科学中， 然而，目前可用的技术局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，从而阻止了从噪声中识别网络激活的一致模式， 网络的标识响应于干预而随时间改变。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。这提供了优于现有技术的许多优点，包括使用高通量自动成像技术分析脑的能力，基于脑切片中的特定细胞群在whoe动物中的激活模式识别脑切片中的特定细胞群以用于电生理学、形态学或分子研究的能力，以及将FACS分选技术应用于分离单个细胞核以用于表观遗传学研究的能力。报告活性的两个时间点可以间隔至少一周，从而可以分析对延长的行为或药理学干预有反应的回路变化和靶细胞群。这应该有助于识别大脑对这些疗法的反应的关键变化。

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.