Isolation of brain mitochondria from neonatal mice

Xiaoyang Wang, Anna-Lena Leverin, Wei Han, Changlian Zhu, Bengt Johansson, Etienne Jacotot, Vadim Ten, Neil Sims, Henrik Hagberg

    Research output: Contribution to journalArticlepeer-review

    20 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)1253-1261
    Number of pages9
    JournalJournal of Neurochemistry
    Volume119
    Issue number6
    DOIs
    Publication statusPublished - Dec 2011

    Keywords

    • brain
    • mitochondrial isolation
    • mouse
    • neonatal

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