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Physiological oxygen level is critical for modeling neuronal metabolism in vitro
Author(s) -
Zhu Jing,
Aja Susan,
Kim EunKyoung,
Park Min Jung,
Ramamurthy Santosh,
Jia Junling,
Hu Xueying,
Geng Ping,
Ronnett Gabriele V.
Publication year - 2012
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.22765
Subject(s) - ampk , glycolysis , metabolism , oxygen , in vitro , intracellular , biochemistry , mitochondrion , microbiology and biotechnology , protein kinase a , carbohydrate metabolism , reactive oxygen species , viability assay , biology , cellular respiration , chemistry , kinase , organic chemistry
In vitro models are important tools for studying the mechanisms that govern neuronal responses to injury. Most neuronal culture methods employ nonphysiological conditions with regard to metabolic parameters. Standard neuronal cell culture is performed at ambient (21%) oxygen levels, whereas actual tissue oxygen levels in the mammalian brain range from 1% to 5%. In this study, we examined the consequences of oxygen level on the viability and metabolism of primary cultures of cortical neurons. Our results indicate that physiological oxygen level (5% O 2 ) has a beneficial effect on cortical neuronal survival and mitochondrial function in vitro. Moreover, oxygen level affects metabolic fluxes: glucose uptake and glycolysis was enhanced at physiological oxygen level, whereas glucose oxidation and fatty acid oxidation were reduced. Adenosine monophosphate‐activated protein kinase (AMPK) was more activated in 5% O 2 and appears to play a role in these metabolic effects. Inhibiting AMPK activity with compound C decreased glucose uptake, intracellular ATP level, and viability in neurons cultured in 5% O 2 . These data indicate that oxygen level is an important parameter to consider when modeling neuronal responses to stress in vitro. © 2011 Wiley Periodicals, Inc.