Loose-patch-juxtacellular recording in vivo - A method for functional characterization and labeling of neurons in macaque V1
Loose-patch-juxtacellular recording in vivo - A method for functional characterization and labeling of neurons in macaque V1
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DOI：
10.1016/j.jneumeth.2006.02.004
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发表时间：
2006-09-30
影响因子：
3
通讯作者：
Hawken, M. J.
中科院分区：
文献类型：
--
作者：
Joshi, S.;Hawken, M. J.
我们描述了一种方法，该方法使用一个修改版本的细胞标记[Pinault D.一种新的单细胞染色程序在体内进行电生理控制下：形态功能特征的aptacellarily标记的丘脑细胞和其他中枢神经元与biocytin或neurobiotin。J Neurosci Meth 1996; 65：113 - 36]，其允许我们在猕猴VI中在体内功能上表征并随后标记单个神经元。该方法通常适用于急性体内制备。当电极附着在细胞膜上时，用贴片电极进行细胞外记录。最初，一个“盲”搜索方法被用作一个指南，以获得一个细胞连接的配置，我们称之为一个松散的补丁（LP）。神经元的感受野特性的功能特征，神经元被标记，然后确认表征，所有在LP配置。我们描述的方法比其他方法有许多优点。首先，我们已经发现，我们可以获得稳定的细胞外记录数小时，使我们能够作出一个相对全面的视觉功能表征神经元的感受野特性。第二，由于电极与细胞紧密贴壁，我们获得了细胞外尖峰的良好隔离。第三，该方法提供了标记，该标记给出了存活数天的完整树突和轴突填充，这是急性灵长类动物实验中的重要特征。第四，标记和重建神经元的体内方法给出了神经元的完整三维结构，包括其皮质内轴突乔木。这些特征克服了研究神经元形态学的既定方法的已知限制，包括高尔基染色（使用成人组织时受到限制）和体外全细胞方法（由于切片厚度有限，轴突填充不完全）。它们还克服了已建立的功能-形态学联合研究方法的已知限制，即体内细胞内记录。改良的细胞外方法为通过细胞外记录和随后的细胞内标记进行表征的困难方法提供了可靠的替代方法[安德森JC，Martin KAC，Whitteridge D.猕猴纹状皮质神经元的形态、功能和皮质内投射。大脑皮层1993; 3：412 - 20]。我们表明，该方法可用于记录在一系列的深度，通过V1皮层允许采样的神经元在不同的层和功能亚群。链接，然后可以与现有的知识的解剖组织的VI，其中发现的各种形态类的神经元，它们的功能连接和视觉响应特性。(c)2006 Elsevier B.V.保留所有权利。
We describe a method that uses a modified version of juxtacellular labeling [Pinault D. A novel single-cell staining procedure performed in vivo under electrophysiological control: morpho-functional features of juxtacellularly labeled thalamic cells and other central neurons with biocytin or neurobiotin. J Neurosci Meth 1996;65:113-36], which allows us to functionally characterize and subsequently label single neurons in vivo in macaque VI. The method is generally applicable in acute in vivo preparations. Extracellular recording is made with a patch electrode when the electrode is attached to the cell membrane. Initially a 'blind' search method is used as a guide to obtaining a cell attached configuration that we refer to as a loose-patch (LP). The neuron's receptive field properties are functionally characterized, the neuron is labeled and then characterization is confirmed, all in the LP configuration. There are a number of advantages of the method that we describe over other methods. First, we have found that we can obtain stable extracellular recordings for periods of hours that enable us to make a relatively comprehensive visual functional characterization of a neuron's receptive field properties. Second, because the electrode is closely apposed to the cell we obtain excellent isolation of the extracellular spike. Third, the method provides labeling that gives complete dendritic and axonal filling that survives over a number of days, which is an important feature in acute primate experiments. Fourth, the in vivo method of labeling and reconstructing neurons gives complete three-dimensional structure of the neuron including its intra-cortical axonal arbor. These features overcome known limits of the established methods of studying neuronal morphology including the Golgi stain (limited when adult tissue is used) and in vitro whole cell methods (incomplete axonal filling due to limited slice thickness). They also overcome the known limits of the established method of combined function-morphology studies i.e. intracellular recording in vivo. The modified juxtacellular method provides a reliable alternative to the difficult method of characterization by extracellular recording and subsequent intracellular labeling [Anderson JC, Martin KAC, Whitteridge D. Form, function and intracortical projections of neurons in the striate cortex of the monkey Macacus nemestrinus. Cerebral Cortex 1993;3:412-20]. We show the method can be used to record at a range of depths through V 1 cortex allowing for sampling of neurons in the different layers and functional subpopulations. Links can then be made with existing knowledge about the anatomical organization of V I, the various morphological classes of neurons found therein, their functional connectivity and visual response properties. (c) 2006 Elsevier B.V. All rights reserved.