Precursors of glutamic acid nitrogen in primary neuronal cultures: studies with 15N

Neurochemical Research
M YudkoffL Hertz

Abstract

We utilized gas chromatography-mass spectrometry to study the transfer of 15N from [2-15N]glutamine, [15N]leucine, [15N]alanine, or 15NH4Cl to [15N]glutamate and [15N]aspartate in cultured cerebrocortical GABA-ergic neurons from the mouse. Initial rates of 15N appearance (atom % excess) were somewhat higher with 2mM [2-15N]glutamine as a precursor than with 1mM [15N]leucine or 1mM [15N]alanine, but initial net formation (nmol [15N]glutamate/mg protein.min-1) was roughly comparable with all precursors. At steady-state 15N labeling was about two times greater with 2mM [2-15N]glutamine as precursor. The subsequent transfer of 15N from glutamate to aspartate was extremely rapid, the labelling pattern of these two amino acid pools being virtually indistinguishable. We observed little reductive amination of 2-oxo-glutarate to yield [15N]glutamate in the presence of 0.3mM 15NH4Cl. Reductive amination through glutamate dehydrogenase was much more prominent at a concentration of 3.0mM 15NH4Cl. Glutamate formation via reductive amination was unaffected by inclusion of 1mM 2-oxo-glutarate in the incubation medium. These results indicate that glutamate synthesis in cultured GABA-ergic neurons is derived not only from the glutaminase reacti...Continue Reading

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Citations

Oct 1, 1992·Journal of Neuroscience Research·L HertzA Schousboe
Aug 1, 1993·Journal of Neuroscience Research·N WestergaardA Schousboe
Apr 25, 2000·Archives of Biochemistry and Biophysics·M MacK A Nałecz
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May 16, 2013·Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism·Sherry DadsetanHelle S Waagepetersen
Jun 22, 2006·Journal of Neurochemistry·Lasse K BakHelle S Waagepetersen
Mar 1, 1994·Journal of Neurochemistry·M YudkoffI Nissim
Nov 1, 1995·The Annals of Pharmacotherapy·D F WeltyJ D Rothstein
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Mar 29, 2000·The Journal of Nutrition·Y Daikhin, M Yudkoff
Jun 22, 2000·The Journal of Comparative Neurology·J LevensonA Eskin
Jun 3, 2005·The Journal of Nutrition·Marc YudkoffItzhak Nissim

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