Isolation of brain mitochondria from neonatal mice.
Isolation of brain mitochondria from neonatal mice.
复制标题
新生小鼠脑线粒体的分离
DOI：
10.1111/j.1471-4159.2011.07525.x
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发表时间：
2011-12
影响因子：
4.7
通讯作者：
Hagberg H
中科院分区：
文献类型：
--
作者：
Wang X;Leverin AL;Han W;Zhu C;Johansson BR;Jacotot E;Ten VS;Sims NR;Hagberg H
线粒体是许多形式的细胞死亡的关键因素，包括由新生儿缺氧缺血性脑损伤引起的死亡。小鼠在脑损伤的研究中越来越受欢迎，但很少有报告评估新生小鼠大脑的线粒体分离程序。利用呼吸活性、标记酶、免疫印迹和电子显微镜的评估，我们比较了以前发表的从新生小鼠脑中分离线粒体的方法(方法A)与适用于成年大鼠(方法B)和新生大鼠(方法C)的方法。这三个程序都使用Percoll密度梯度离心法作为分离的关键步骤，但在分馏程序和分馏过程中使用的溶液的许多方面有所不同。方法A和B都能产生高度浓缩的具有高呼吸活性的连接良好的线粒体组分。方法C的组份对呼吸特性的保存性较差，并被其他亚细胞成分污染较多。方法A的优点是更快速，产生更大的线粒体产量，使其适用于常规应用。然而，方法B产生的线粒体受到突触体和相关胞浆成分的污染较少，这适合于对线粒体提纯有更高要求的研究。
Mitochondria are key contributors to many forms of cell death including those resulting from neonatal hypoxic-ischemic brain injury. Mice have become increasingly popular in studies of brain injury, but there are few reports evaluating mitochondrial isolation procedures for the neonatal mouse brain. Using evaluation of respiratory activity, marker enzymes, western blotting and electron microscopy, we have compared a previously published procedure for isolating mitochondria from neonatal mouse brain (method A) with procedures adapted from those for adult rats (method B) and neonatal rats (method C). All three procedures use Percoll density gradient centrifugation as a key step in the isolation but differ in many aspects of the fractionation procedure and the solutions used during fractionation. Methods A and B both produced highly enriched fractions of well-coupled mitochondria with high rates of respiratory activity. The fraction from method C exhibited less preservation of respiratory properties and was more contaminated with other subcellular components. Method A offers the advantage of being more rapid and producing larger mitochondrial yields making it useful for routine applications. However, method B produced mitochondria that were less contaminated with synaptosomes and associated cytosolic components that suits studies that have a requirement for higher mitochondrial purification.
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影响因子：
1.9
作者：
Hagberg H;Mallard C;Rousset CI;Xiaoyang Wang
通讯作者：
Xiaoyang Wang
影响因子：
2.9
作者：
Hansson, MJ;Persson, T;Elmér, E
通讯作者：
Elmér, E
影响因子：
14.8
作者：
Frezza, Christian;Cipolat, Sara;Scorrano, Luca
通讯作者：
Scorrano, Luca
影响因子：
2.9
作者：
Caspersen, Casper S.;Sosunov, Alexander;Ten, Vadim S.
通讯作者：
Ten, Vadim S.
DOI：
10.1523/jneurosci.5832-08.2009
发表时间：
2009-02-25
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Wang X;Carlsson Y;Basso E;Zhu C;Rousset CI;Rasola A;Johansson BR;Blomgren K;Mallard C;Bernardi P;Forte MA;Hagberg H
通讯作者：
Hagberg H