1. In rats in a variety of nutritional states, the adipose tissue clearing factor lipase activity is strongly, positively correlated with fat-cell triglyceride fatty acid uptake. 2. In the same animals, muscle clearing factor lipase activity is inversely correlated with the activity of the enzyme in adipose tissue and with the plasma insulin concentration. 3. In starved animals that are given glucose, adipose tissue clearing factor lipase activity is positively correlated with the plasma insulin concentration. 4. The effect of changes in nutritional status on the activity of clearing factor lipase in rat post-heparin plasma depends on the heparin dosage used. The administration of glucose, but not of fructose or sucrose, to starved rats alters the response to heparin injection towards that found in rats in the fed state.
1. When fat-cells are isolated from the epididymal adipose tissue of 24h-starved rats and incubated at 25 degrees C in the presence of dialysed serum, glucose, insulin, amino acids and heparin, the total clearing-factor lipase acitivity of the incubation system increases progressively over a period of several hours. 2. All of the increase in activity is accounted for by the appearance of enzyme in the appearance of enzyme in the incubation medium and the fat-cell activity does not change significantly. Cycloheximids, at a concentration that prevents protein synthesis, does not affect the appearance of enzyme in the incubation medium, but the fat-cell enzyme activity is decreased in its presence. 3. The magnitude of the increase in total clearing factor lipase activity is unaffected by the omission of heparin from the medium. However, less enzyme is extracted in tis absence and the fat-cell activity increases. Cycloheximide again only affects the rise in cell activity and does not alter the activity in the incubation medium. 4. When serum in the incubation medium is replaced by casein, the distribution of enzyme between the cells and the medium is changed, but the magnitudes of the increases in total enzyme activity are similar. 5. These characteristics of the clearing-factor lipase response of isolated fat-cells differ in several respects from those observed earlier with intact adipose tissue from 24h-starved rats (Robinson & Wing, 1971; Cryer et al., 1973). The differences could be due, in part, to changes in the relative amounts of two different molecular forms of the enzyme that occur during the isolation of the fat-cells.
When isolated fat-cells are incubated at 25 degrees C in serum-based media containing glucose, insulin and heparin, the rise that occurs in the clearing-factor lipase activity of the incubation medium is inhibited by colchicine. The rise in the fat-cell clearing-factor lipase activity that occurs during similar incubations in the absence of heparin is not affected by colchicine.
The rise in adipose-tissue clearing-factor lipase activity that results from feeding glucose to starved rats cannot be duplicated by giving equicaloric amounts of fructose or sucrose. An inability of the administered fructose and sucrose to raise the plasma insulin concentration probably accounts for this failure in enzyme response.