The mechanism responsible for insulin resistance during pregnancy remains unclear. Considerable evidence indicates that insulin receptor substrate-1 could play an important role in insulin sensitivity. It seems possible that the gestational hormonal milieu could affect insulin receptor substrate-1. In the present study, measurements of tyrosine phosphorylation and protein content of insulin receptor substrate-1 and gene expression in the liver, skeletal muscle and adipose tissue in the rat indicated that, during pregnancy, significant changes occurred in these parameters. We found in early gestation that muscle and adipose tissue were highly sensitive to insulin action, because the phosphorylation of insulin receptor substrate-1 is greater than in late gestation. However, in late gestation the tissue most sensitive to insulin action, reflecting insulin receptor substrate-1 phosphorylation, was the liver. Our hypothesis was that these results are connected with the changes in concentrations of estradiol and progesterone observed during pregnancy. It was concluded that the present findings demonstrate that different concentrations of gestational hormones play an important role in insulin sensitivity in this period, and that each tissue responds in the most appropriate manner to guarantee the gestation in its entirety, controlling the phosphorylation of insulin receptor substrate-1 in response to insulin receptor activation.