The use of tritiated water to measure absolute rates of hepatic glycogen synthesis

Abstract

Glucogen synthesis in rat liver in vivo was measured by the incorporation of 3H from 3H2O into glycogen. In meal-fed rats incorporation and the incorporation of 3H into glycogen was linear up to 100 min. Before feeding glycogen concentration and the incorporation of 3H were both low; and both rose on feeding to give maximal values after 2-3h. The glycogen concentration was maintained for a further 5h but the incorporation of 3H rapidly declined to pre-feeding values. This shows that glycogen turnover was low in the post-prandial rat. Streptozotocin diabetes decreased the rise in glycogen concentration on feeding and had a similar effect on 3H2O incorporation. Both effects were reversed by insulin administration. The number of 3H atoms incorporated per glycogen glucose moiety formed in biosynthetic experiments (2.84 +/- 0.47) was relatively constant and allowed absolute biosynthetic rates to be calculated. Degradation of glucose from glycogen labelled by 3H2O showed that most of the 3H was located at C-2 and C-5. The incorporation would arise by rapid equilibration of hexose phosphates through phosphoglucose isomerase, transaldolase and triose phosphate isomerase.