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. 1966 Aug;100(2):407–419. doi: 10.1042/bj1000407

Measurement of concentrations of metabolites in adipose tissue and effects of insulin, alloxan-diabetes and adrenaline

R M Denton 1, R E Yorke 1,*, P J Randle 1
PMCID: PMC1265150  PMID: 5968539

Abstract

1. Methods are described for the extraction and assay of ATP, ADP, AMP, glucose 6-phosphate, l-glycerol 3-phosphate and citrate in rat epididymal adipose tissue incubated in vitro for 1hr. At this time of incubation rates of glucose uptake and outputs of glycerol, free fatty acids, lactate and pyruvate were shown to be constant. 2. In fat pads incubated in medium containing glucose (3mg./ml.) and albumin (20mg./ml.) the concentrations (in mμmoles/g. wet wt.) were: ATP, 70; ADP, 36; AMP, 9·0; glucose 6-phosphate, 3·0; l-glycerol 3-phosphate, 3·3; citrate, 8·1. 3. The volume of intracellular water calculated from ([3H]water space–[14C]sorbitol space), ([14C]urea space–inulin space) and (weight loss on drying–[14C]sorbitol space) was 1·4ml./100g. wet wt. of tissue. The intracellular volume was not changed by insulin, alloxan-diabetes or adrenaline. 4. When compared in terms of μmoles/ml. of intracellular water the concentration of ATP in adipose tissue was less than in heart and diaphragm muscles. The concentrations of ADP and AMP were greater both in absolute terms and relative to ATP. Insulin, alloxan-diabetes and adrenaline had no significant effects on the concentrations of the adenine nucleotides in adipose tissue. 5. The concentration of glucose 6-phosphate was increased by insulin and lowered by alloxan-diabetes and adrenaline. The concentration of l-glycerol 3-phosphate was increased by insulin, unchanged by alloxan-diabetes and lowered by adrenaline. The concentration of citrate was increased by adrenaline and alloxan-diabetes and unchanged by insulin. 6. The effect of glucose concentration in the medium on rates of glucose uptake in adipose tissue from normal rats and alloxan-diabetic rats was investigated. The Ku of glucose uptake was 29–44mg./100ml. and the Vmax. was 0·77mg./g. wet wt. of tissue/hr. Insulin increased the Vmax. and alloxan-diabetes diminished it, but neither agent significantly altered the Ku. 7. The significance of these results in relation to control of metabolism of adipose tissue is discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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