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Title: 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 whichmore » occur when cells are grown in culture. 59 references. 31 figures.« less
Authors:
Publication Date:
OSTI Identifier:
6020082
Report Number(s):
LBL-18628
ON: DE85004604
DOE Contract Number:
AC03-76SF00098
Resource Type:
Technical Report
Resource Relation:
Other Information: Thesis
Research Org:
Lawrence Berkeley Lab., CA (USA)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 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