A simple method for isolating rat brain mitochondria with high metabolic activity: Effects of EDTA and EGTA

被引：30

作者：

Iglesias-Gonzalez, Javier ^{[1
]}

Sanchez-Iglesias, Sofia ^{[1
]}

Beiras-Iglesias, Andres ^{[2
]}

Soto-Otero, Ramon ^{[1
]}

Mendez-Alvarez, Estefania ^{[1
]}

机构：

[1] Univ Santiago de Compostela, Fac Med, Dept Biochem & Mol Biol, Lab Neurochem, Santiago De Compostela 15782, Spain

[2] Univ Santiago de Compostela, Fac Med, Dept Morphol Sci, Histol Lab, Santiago De Compostela 15782, Spain

来源：

JOURNAL OF NEUROSCIENCE METHODS | 2013年 / 213卷 / 01期

关键词：

Mitochondria; Brain; EDTA; EGTA; Isolation procedure; Mitochondrial respiration; Oxidative stress; LIVER MITOCHONDRIA; RESPIRATION; SYNAPTOSOMES; DISEASE; OXIDASE;

D O I：

10.1016/j.jneumeth.2012.12.005

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Isolated mitochondria are widely used in metabolic and oxidative stress studies for neurodegenerative diseases. In the present work, the influence of EGTA and EDTA has been tested on a sucrose-based differential centrifugation protocol in order to establish the optimal concentrations to be used in this process. Our results showed alterations in both active and resting respiration, which were dependent on both the addition of EDTA or EGTA to the isolation buffer and the chelator concentration used. However, the addition of chelator to the isolation medium does not modify the mitochondria structure as assessed by both distribution of biological markers and electron micrography in the final pellet. Our results endorse this protocol as the method of choice for metabolic and oxidative stress experiments with fresh isolated rat brain mitochondria. (c) 2012 Elsevier B.V. All rights reserved.

引用

页码：39 / 42

页数：4

共 20 条

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