Measuring dendritic distribution of membrane proteins
Measuring dendritic distribution of membrane proteins
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DOI：
10.1016/j.jneumeth.2006.03.014
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发表时间：
2006-09-30
影响因子：
3
通讯作者：
Heckman, C. J.
中科院分区：
文献类型：
--
作者：
Ballou, Edmund W.;Smith, W. Bryan;Heckman, C. J.
神经元在树突树中完成大部分整合工作，而脊髓运动神经元的树突树是所有细胞中最大的。电兴奋性强烈影响枝晶膜的性质，这是很难直接测量。我们描述了一种测量离子通道膜密度沿沿着树枝状轨迹分布的方法。该方法结合了标准的免疫组织化学与重建程序的大规模和小规模的光学显微镜。然后，为Matlab编写的软件提取靶离子通道与靶染料注射细胞的共定位，并计算每平方微米细胞表面积的相对通道密度，作为与细胞体的距离的函数。该技术可用于量化任何免疫反应性部分的定位和分布，并且该软件提供了用于灵敏度分析的灵活工具，以验证用于选择阈值的免疫学。(c)2006 Elsevier B.V.保留所有权利。
Neurons perform much of their integrative work in the dendritic tree, and spinal motoneurons have the largest tree of any cell. Electrical excitability is strongly influenced by dendrite membrane properties, which are difficult to measure directly. We describe a method to measure the distribution of ion channel membrane densities along dendritic trajectories. The method combines standard immunohistochemistry with reconstruction procedures for both large-scale and small-scale optical microscopy. Software written for Matlab then extracts the colocalization of the target ion channel with the target dye injected cell, and calculates the relative channel density per square micron of cell surface area, as a function of distance from the cell body. The technique can be used to quantify the localization and distribution of any immunoreactive moiety, and the software provides a flexible vehicle for sensitivity analysis, to validate heuristics for selecting thresholds. (c) 2006 Elsevier B.V. All rights reserved.