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首页> 外文期刊>Journal of Neuroscience Methods >Method for functional study of mitochondria in rat hypothalamus.
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Method for functional study of mitochondria in rat hypothalamus.

机译:大鼠下丘脑线粒体功能研究的方法。

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摘要

Different roles of mitochondria in brain function according to brain area are now clearly emerging. Unfortunately, no technique is yet described to investigate mitochondria function in specific brain area. In this article, we provide a complete description of a procedure to analyze the mitochondrial function in rat brain biopsies. Our two-step method consists in a saponin permeabilization of fresh brain tissues in combination with high-resolution respirometry to acquire the integrated respiratory rate of the biopsy. In the first part, we carefully checked the mitochondria integrity after permeabilization, defined experimental conditions to determine the respiratory control ratio (RCR), and tested the reproducibility of this technique. In the second part, we applied our method to test its sensitivity. As a result, this method was sensitive enough to reveal region specificity of mitochondrial respiration within the brain. Moreover, we detected physiopathological modulation of the mitochondrial function in the hypothalamus. Thus this new technique that takes all cell types into account, and does not discard or select any mitochondria sub-population is very suitable to analyze the integrated mitochondrial respiration of brain biopsies.
机译:线粒体在脑功能中的不同作用(根据大脑区域)现已清楚地显现出来。不幸的是,还没有描述研究特定脑区域中线粒体功能的技术。在本文中,我们提供了在大鼠脑活检中分析线粒体功能的程序的完整说明。我们的两步方法包括将新鲜脑组织的皂素通透化与高分辨率呼​​吸测定法相结合,以获取活检的综合呼吸频率。在第一部分中,我们仔细检查了透化后的线粒体完整性,定义了确定呼吸控制率(RCR)的实验条件,并测试了该技术的可重复性。在第二部分中,我们应用了我们的方法来测试其灵敏度。结果,这种方法足够灵敏,可以揭示大脑中线粒体呼吸的区域特异性。此外,我们检测到下丘脑线粒体功能的生理病理调节。因此,这种将所有细胞类型都考虑在内且不会丢弃或选择任何线粒体亚群的新技术非常适合分析脑活检的综合线粒体呼吸。

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