title: Isolation of brain mitochondria from neonatal mice.
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.
authors: Perinatal Center, Institute of Neuroscience and Physiology, University of Gothenburg, Gothenburg, Sweden. xiaoyang.wang@fysiologi.gu.se
authors: Xiaoyang
authors: X
authors: Wang
authors: Anna-Lena
authors: AL
authors: Leverin
authors: Wei
authors: W
authors: Han
authors: Changlian
authors: C
authors: Zhu
authors: Bengt R
authors: BR
authors: Johansson
authors: Etienne
authors: E
authors: Jacotot
authors: Vadim S
authors: VS
authors: Ten
authors: Neil R
authors: NR
authors: Sims
authors: Henrik
authors: H
authors: Hagberg
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000648:ultrastructure*; D003576:Electron Transport Complex IV / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D000244:Adenosine Diphosphate; D007770:L-Lactate Dehydrogenase; D003576:Electron Transport Complex IV