Effect of nerve growth factor on the synthesis of amino acids in PC12 cells
- Univ. of Maryland, Baltimore (USA)
Radioactive short-chain fatty acids preferentially label glutamine relative to glutamate in brain due to compartmentation of glutamine and glutamate. To determine whether this phenomenon occurs in a single cell culture model, we examined the effect of fatty acid chain length on the synthesis as well as pool size of selected amino acids in rat pheochromocytoma PC12 cells, a cell culture model of the large glutamate compartment in neurons. Intracellular 14C-amino acids were quantitated by HPLC, and the incorporation of (U-14C)-glucose, (1-14C)-butyrate, (1-14C)-octanoate, and (1-14C)-palmitate into five amino acids was measured in native and NGF-treated PC12 cells. NGF pretreatment decreased the intracellular concentration of amino acids as did addition of fatty acids but these effects were not additive. Specific activities of amino acids in native cells labelled by 14C-octanoate were 1,300 DPM/nmol, 490 DPM/nmol, 200 DPM/nmol, and 110 DPM/nmol for glutamate, aspartate, glutamine, and serine, respectively. No radioactivity was detected in alanine. Similar specific activities were noted when 14C-butyrate was the precursor; however, there was at least 5-fold less if 14C-palmitate was the precursor. Pretreatment of cells with NGF decreased the specific activity of amino acids by 25-65%. Specific activities of amino acids synthesized from 14C-glucose decreased in the following order: glutamate, 1,640 DPM/nmol; aspartate, 1,210 DPM/nmol; alanine, 580 DPM/nmol; glutamine, 275 DPM/nmol; and serine, 80 DPM/nmol for native cells. NGF pretreatment decreased the specific activities of glutamate and glutamine, but not of the other 3 amino acids. The preferred precursor for glutamate synthesis in native PC12 cells was glucose followed by octanoate, butyrate and palmitate (16:6:3:1).
- OSTI ID:
- 5228833
- Journal Information:
- Journal of Neuroscience Research; (USA), Journal Name: Journal of Neuroscience Research; (USA) Vol. 22:4; ISSN JNRED; ISSN 0360-4012
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ADRENAL GLANDS
ALDEHYDES
AMINO ACIDS
ANIMAL CELLS
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
BODY
BUTYRIC ACID
CARBOHYDRATES
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CHROMATOGRAPHY
ENDOCRINE GLANDS
ENZYMES
GLANDS
GLUCOSE
GLUTAMIC ACID
GROWTH FACTORS
HEXADECANOIC ACID
HEXOSES
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LIGASES
LIQUID COLUMN CHROMATOGRAPHY
MITOGENS
MONOCARBOXYLIC ACIDS
MONOSACCHARIDES
NERVES
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PRECURSOR
PROTEINS
REACTION KINETICS
SACCHARIDES
SEPARATION PROCESSES
SYNTHESIS
TRACER TECHNIQUES
TUMOR CELLS