Glucokinase (EC 220.127.116.11) first appears in rat liver two weeks after birth and increases rapidly after weaning on to a high-carbohydrate diet. We investigated the role of triiodothyronine and retinoic acid in the absence of insulin on the first expression of the glucokinase gene in primary cultures of hepatocytes from 10 day-old rats. These two hormones were able to induce a rapid accumulation of liver glucokinase mRNA, secondarily to a stimulation of gene transcription during the first 24 h of culture. Moreover, the effects of individual hormones were not additive. Finally, glucokinase mRNA stability was not modified by these hormones. This suggests that triiodothyronine and retinoic acid act on glucokinase gene at the transcriptional.