Demonstration of NeuronGlia Transfer of Precursor

Authors: Renata Leke Lasse K Bak Arne Schousboe Helle S Waagepetersen

Publish Date: 2008/08/16

Volume: 33, Issue: 12, Pages: 2629-2635

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Abstract

Cocultures of neurons and astrocytes were prepared from dissociated embryonic mouse cerebral cortex and cultured for 7 days To investigate if these cultures may serve as a functional model system to study neuronglia interaction with regard to GABA biosynthesis the cells were incubated either in media containing U13Cglutamine 01 03 and 05 mM or 1 mM acetate plus 25 mM glucose plus 1 mM lactate In the latter case one of the 3 substrates was uniformly 13C labeled Cellular contents and 13C labeling of glutamate GABA aspartate and glutamine were determined in the cells after an incubation period of 25 h The GABA biosynthetic machinery exhibited the expected complexity with regard to metabolic compartmentation and involvement of TCA cycle activity as seen in other culture systems containing GABAergic neurons Metabolism of acetate clearly demonstrated glial synthesis of glutamine and its transfer to the neuronal compartment It is concluded that this coculture system serves as a reliable model in which functional and pharmacological aspects of GABA biosynthesis can be investigatedThe expert secretarial assistance of Ms Hanne Danø and the skilful technical assistance of Ms Ann Lene Vigh are highly appreciated The experimental work has been supported by grants from the Danish Medical Research Council 22040314 and 271070267 and the Lundbeck Hørslev and Novo Nordisk Foundations A travel grant to RL from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Capes is coordially acknowledged

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