Chronic cellular imaging of mouse visual cortex during operant behavior and passive viewing.
Chronic cellular imaging of mouse visual cortex during operant behavior and passive viewing.
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DOI：
10.3389/fncel.2010.00003
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发表时间：
2010
影响因子：
5.3
通讯作者：
Reid RC
中科院分区：
文献类型：
--
作者：
Andermann ML;Kerlin AM;Reid RC
哺乳动物新皮质中的邻近神经元呈现出细胞类型和连接模式的高度多样性。这种多样性对计算的重要性尚不清楚。虽然视觉皮层的细胞外记录研究对神经活动的行为调节进行了非常丰富的描述，但需要新的方法来剖析特定回路元件在引导视觉感知中的作用。在此，我们描述了一种在清醒小鼠视觉皮层中对主动辨别和被动观看视觉刺激期间的神经活动进行三维细胞成像的方法。头部固定的小鼠在初步掌握任务后，每天能进行数百次稳健的辨别试验。为了在操作行为期间以单次试验分辨率且最小化伪影记录多个神经元，我们构建了一台用于双光子钙成像的灵敏显微镜，它能够在三维空间中快速追踪神经元。我们使用合成的和基因编码的钙指示剂，证明了在数小时、数天和数周的辨别行为过程中细胞钙活动的稳定记录。当与小鼠的分子和基因技术（例如，细胞类型特异性转基因标记）相结合时，这种方法能够在体内识别神经元类别。对邻近的不同类别神经元进行生理测量将丰富我们对皮质微回路不同元件在引导感觉感知和知觉学习中的协同作用的理解。此外，我们的方法提供了一种高通量、慢性的体内检测方法，用于检测行为对细胞活动的影响，该方法适用于多种神经系统疾病的小鼠模型。
Nearby neurons in mammalian neocortex demonstrate a great diversity of cell types and connectivity patterns. The importance of this diversity for computation is not understood. While extracellular recording studies in visual cortex have provided a particularly rich description of behavioral modulation of neural activity, new methods are needed to dissect the contribution of specific circuit elements in guiding visual perception. Here, we describe a method for three-dimensional cellular imaging of neural activity in the awake mouse visual cortex during active discrimination and passive viewing of visual stimuli. Head-fixed mice demonstrated robust discrimination for many hundred trials per day after initial task acquisition. To record from multiple neurons during operant behavior with single-trial resolution and minimal artifacts, we built a sensitive microscope for two-photon calcium imaging, capable of rapid tracking of neurons in three dimensions. We demonstrate stable recordings of cellular calcium activity during discrimination behavior across hours, days, and weeks, using both synthetic and genetically encoded calcium indicators. When combined with molecular and genetic technologies in mice (e.g., cell-type specific transgenic labeling), this approach allows the identification of neuronal classes in vivo. Physiological measurements from distinct classes of neighboring neurons will enrich our understanding of the coordinated roles of diverse elements of cortical microcircuits in guiding sensory perception and perceptual learning. Further, our method provides a high-throughput, chronic in vivo assay of behavioral influences on cellular activity that is applicable to a wide range of mouse models of neurologic disease.
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影响因子：
2.5
作者：
Boudreau, C. Elizabeth;Williford, Tori H.;Maunsell, John H. R.
通讯作者：
Maunsell, John H. R.
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通讯作者：
Kerr, Jason N. D.
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作者：
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