Nitrogen disruption of synaptoneurosomes: an alternative method to isolate brain mitochondria.

TitleNitrogen disruption of synaptoneurosomes: an alternative method to isolate brain mitochondria.
Publication TypeJournal Article
Year of Publication2004
JournalJournal of neuroscience methods
Volume137
Issue2
Pagination299-303
Date Published2004
ISSN0165-0270
Abstract

Mitochondria are known to be localized in synaptic and non-synaptic compartments in the brain. Synaptoneurosomes, which contain high numbers of mitochondria, may act as a major contaminant of currently used isolation techniques. Currently, there is no method employed to successfully disrupt synaptoneurosomes and isolate both synaptic and non-synaptic mitochondria without structural or functional damage. A novel method is reported here for disruption of synaptoneurosomes and isolation of total brain mitochondria from synaptic and non-synaptic sources using a nitrogen decompression technique. Nitrogen gas was dissolved into crude mitochondrial preparations and maintained under constant, moderate pressure. After a short incubation, the pressure was released causing the nitrogen to come out of solution as growing bubbles, which ruptures cellular and synaptoneurosomal membranes. Mitochondria isolated using this rapid technique were bioenergetically competent and exhibited functional characteristics comparable to mitochondria isolated using traditional techniques. This nitrogen decompression technique will allow for further characterization of synaptic pools of mitochondria, which are almost exclusively neuronal in origin.

URLhttps://linkinghub.elsevier.com/retrieve/pii/S0165027004001001
DOI10.1016/j.jneumeth.2004.02.028
Short TitleJ Neurosci Methods
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