Nitrogen disruption of synaptoneurosomes: An alternative method to isolate brain mitochondria

Maile R. Brown; Patrick G. Sullivan; Kristina A. Dorenbos; Edward A. Modafferi; James W. Geddes; Oswald Steward

doi:10.1016/j.jneumeth.2004.02.028

Nitrogen disruption of synaptoneurosomes: An alternative method to isolate brain mitochondria

Maile R. Brown, Patrick G. Sullivan, Kristina A. Dorenbos, Edward A. Modafferi, James W. Geddes, Oswald Steward

Research output: Contribution to journal › Article › peer-review

94 Scopus citations

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.

Original language	English
Pages (from-to)	299-303
Number of pages	5
Journal	Journal of Neuroscience Methods
Volume	137
Issue number	2
DOIs	https://doi.org/10.1016/j.jneumeth.2004.02.028
State	Published - Aug 30 2004

Bibliographical note

Funding Information:
The authors would like to thank Dr. Janet Dubinsky, Dr. Tibor Kristian and Dr. Gary Fiskum for thoughtful insight and advice. M.R.B. is a predoctoral trainee on a National Institutes of Health Training Grant AG00264. This work was supported by a NeoTherapeutics Fellowship (to P.G.S.), the Alzheimer’s Association (to J.W.G.), and by the National Institutes of Health, U.S. Public Health Service grants NS048191 (to P.G.S.), AG10836 (to J.W.G.) and NS12333 (to O.S.).

Keywords

Isolation brain mitochondria
Mitochondria
Nitrogen decompression
Non-synaptic
Synaptic
Total

ASJC Scopus subject areas

General Neuroscience

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10.1016/j.jneumeth.2004.02.028

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title = "Nitrogen disruption of synaptoneurosomes: An alternative method to isolate brain mitochondria",

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.",

keywords = "Isolation brain mitochondria, Mitochondria, Nitrogen decompression, Non-synaptic, Synaptic, Total",

author = "Brown, {Maile R.} and Sullivan, {Patrick G.} and Dorenbos, {Kristina A.} and Modafferi, {Edward A.} and Geddes, {James W.} and Oswald Steward",

note = "Funding Information: The authors would like to thank Dr. Janet Dubinsky, Dr. Tibor Kristian and Dr. Gary Fiskum for thoughtful insight and advice. M.R.B. is a predoctoral trainee on a National Institutes of Health Training Grant AG00264. This work was supported by a NeoTherapeutics Fellowship (to P.G.S.), the Alzheimer{\textquoteright}s Association (to J.W.G.), and by the National Institutes of Health, U.S. Public Health Service grants NS048191 (to P.G.S.), AG10836 (to J.W.G.) and NS12333 (to O.S.). ",

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AU - Geddes, James W.

AU - Steward, Oswald

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N2 - 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.

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Nitrogen disruption of synaptoneurosomes: An alternative method to isolate brain mitochondria

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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