Study of cultured fibroblasts in vivo using NMR
The goal was to study the compartmentation of phosphorylated glycolytic intermediates in intact Chicken Embryo Fibroblasts (CEFs) using /sup 31/P NMR at 109 MHz. A technique for maintaining functional cells at high densities in an NMR magnet is described. Signals were detected from cytoplasmic inorganic phosphate (P/sub i/), ATP, NAD, NADH, phosphorylcholine and phosphorylethanolamine. The effect of external glucose on cytoplasmic pools of phosphates was studied. When cells were perfused with glucose-free medium the rate of glycolysis decreased, the amplitudes of the ATP resonances decreased, and the P/sub i/ intensity increased. The quantity of NMR-detectable P/sub i/ produced was significantly greater than the quantity of NMR-detectable ATP which was lost. Experiments with /sup 32/P labeled P/sub i/ showed that as the concentration of glucose in the medium was increase, the amount of phosphate sequestered in the cells increased. We conclude that there is a pool of P/sub i/ which is not detected by high resolution NMR and that the size of this pool increases as the rate of glycolysis increase. Longtitudinal relaxation times of intracellular phosphates in normal, transformed, and primary CEFs were measured. The results demonstrate that relaxation times of phosphates are sensitive to structural and metabolic changes which occur when cells are grown in culture. 59 references. 31 figures.
- Research Organization:
- Lawrence Berkeley Lab., CA (USA)
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 6020082
- Report Number(s):
- LBL-18628; ON: DE85004604
- Resource Relation:
- Other Information: Thesis
- Country of Publication:
- United States
- Language:
- English
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FIBROBLASTS
GLYCOLYSIS
GLUCOSE
BIOLOGICAL EFFECTS
CATABOLISM
ATP
CELL CULTURES
CHICKENS
CYTOPLASM
EMBRYOS
EXPERIMENTAL DATA
NAD
NUCLEAR MAGNETIC RESONANCE
PHOSPHATES
PHOSPHORUS 32
QUANTITY RATIO
ALDEHYDES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIRDS
CARBOHYDRATES
CELL CONSTITUENTS
CHEMICAL REACTIONS
COENZYMES
CONNECTIVE TISSUE CELLS
DATA
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
FOWL
HEXOSES
INFORMATION
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
METABOLISM
MONOSACCHARIDES
NUCLEI
NUCLEOTIDES
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RESONANCE
SACCHARIDES
SOMATIC CELLS
VERTEBRATES
550501* - Metabolism- Tracer Techniques