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Metabolic alterations induced by ischemia in primary cultures of astrocytes: merging 13 C NMR spectroscopy and metabolic flux analysis
Author(s) -
Amaral Ana I.,
Teixeira Ana P.,
Martens Sanja,
Bernal Vicente,
Sousa Marcos F. Q.,
Alves Paula M.
Publication year - 2010
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2010.06636.x
Subject(s) - metabolic flux analysis , metabolism , ischemia , glycolysis , flux (metallurgy) , citric acid cycle , catabolism , pentose phosphate pathway , biology , biochemistry , metabolic pathway , extracellular , astrocyte , metabolic network , microbiology and biotechnology , chemistry , medicine , endocrinology , organic chemistry , central nervous system
J. Neurochem. (2010) 113 , 735–748. Abstract Disruption of brain energy metabolism is the hallmark of cerebral ischemia, a major cause of death worldwide. Astrocytes play a key role in the regulation of brain metabolism and their vulnerability to ischemia has been described. Aiming to quantify the effects of an ischemic insult in astrocytic metabolism, primary cultures of astrocytes were subjected to 5 h of oxygen and glucose deprivation in a bioreactor. Flux distributions, before and after ischemia, were estimated by metabolic flux analysis using isotopic information and the consumption/secretion rates of relevant extracellular metabolites as constraints. During ischemia and early recovery, 30% of cell death was observed; several metabolic alterations were also identified reflecting a metabolic response by the surviving cells. In the early recovery (∼10 h), astrocytes up‐regulated glucose utilization by 30% and increased the pentose phosphate pathway and tricarboxylic acid cycle fluxes by three and twofold, respectively. Additionally, a two to fivefold enhancement in branched‐chain amino acids catabolism suggested the importance of anaplerotic molecules to the fast recovery of the energetic state, which was corroborated by measured cellular ATP levels. Glycolytic metabolism was predominant in the late recovery. In summary, this work demonstrates that changes in fluxes of key metabolic pathways are implicated in the recovery from ischemia in astrocytes.