Kinetics for changes in enzyme synthesis and mRNA content and hormones required for induction of 6-phosphogluconate dehydrogenase in hepatocytes.
Rats fasted for 2 days were refed a 60% glucose diet for varying periods of time in order to follow the kinetics for changes in 6-phosphogluconate dehydrogenase synthesis and mRNA content. Hepatocytes isolated from control or induced rats were incubated with actinomycin D and the rate of decline in 6-phosphogluconate dehydrogenase mRNA was determined by translating RNA in a nuclease-treated reticulocyte lysate. The half-life for 6-phosphogluconate dehydrogenase mRNA under both of these conditions was about 2 h. Thus, increases in transcription or the processing of nuclear RNA may increase 6-phosphogluconate dehydrogenase mRNA during the dietary induction of this enzyme. Hepatocytes prepared from fasted rats were cultured with 5% serum and various hormones and energy sources. If hepatocytes were isolated from thyroidectomized rats and cultured in serum from a thyroidectomized calf, the 4-fold induction of 6-phosphogluconate dehydrogenase was primarily dependent upon added insulin. In the presence of optimal insulin concentrations (10(-7) M) triiodothyronine slightly stimulated 6-phosphogluconate dehydrogenase induction. The gut hormones somatostatin and secretin had no effect on 6-phosphogluconate dehydrogenase induction in cultured hepatocytes. Hepatocytes cultured in carbohydrate-free medium and 5% serum required added insulin for maximal induction. 8-Br-cGMP did not significantly affect 6-phosphogluconate dehydrogenase induction in hepatocytes either in the presence or absence of added insulin. Dibutyryl cAMP did not alter the time course or extent of 6-phosphogluconate dehydrogenase induction in cultured hepatocytes. We have concluded that under these conditions insulin is a potent signal regulating the levels of 6-phosphogluconate dehydrogenase mRNA and that this induction is not mediated by cyclic nucleotides.
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D050260:Carbohydrate Metabolism; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D007700:Kinetics; D008099:Liver / Q000502:physiology*; D008297:Male; D010734:Phosphogluconate Dehydrogenase / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D014158:Transcription, Genetic
D012333:RNA, Messenger; D003994:Bucladesine; D010734:Phosphogluconate Dehydrogenase; D006152:Cyclic GMP