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首页> 外文期刊>Journal of Neuroscience Methods >Method for in situ detection of the mitochondrial function in neurons.
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Method for in situ detection of the mitochondrial function in neurons.

机译:原位检测神经元线粒体功能的方法。

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摘要

Conventional studies of neuronal mitochondria have been limited to the use of purified preparations of isolated mitochondria, neural cell homogenates, living neurons, or brain slices. However, each technique has several drawbacks. Here, we demonstrate that the neuronal cell's membrane can be effectively permeabilized by saponin-treatment and that these permeabilized neurons can be used for qualitative and quantitative assessments of oxygen consumption in combination with registration of mitochondrial membrane potential and free [Ca(2+)] in the matrix. Under these conditions, the mitochondrial function can be studied without removing the mitochondria from their natural milieu thus avoiding the damage of the associated cytoskeleton and outer membrane. At the same time, the method allows the estimation of the mitochondrial function independently of other processes in the cell, and the easy manipulation of the milieu surrounding the mitochondria. Thus, the presented method offers the opportunity to study the neuronal mitochondrial function in situ and can also be applied to examine the mitochondrial function by other commonly used methods.
机译:神经元线粒体的常规研究仅限于使用分离的线粒体,神经细胞匀浆,活神经元或脑切片的纯化制剂。但是,每种技术都有一些缺点。在这里,我们证明了神经元细胞膜可以通过皂素处理有效地被透化,并且这些被透化的神经元可以结合线粒体膜电位和游离[Ca(2+)]的配比用于定性和定量评估耗氧量。在矩阵中。在这些条件下,无需从自然环境中去除线粒体即可研究线粒体功能,从而避免了相关细胞骨架和外膜的损伤。同时,该方法允许独立于细胞中的其他过程来估计线粒体功能,并易于操纵线粒体周围的环境。因此,所提出的方法提供了就地研究神经元线粒体功能的机会,并且还可以通过其他常用方法应用于检查线粒体功能。

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