A large number of experimental studies have investigated insulin signaling in rats. In these studies different anaesthetics have been used to anaesthetize rats. However, the direct effects of anaesthetics on the regulation of the early steps of insulin action are not known. In the present study, we investigated the effect of thiopental, pentobarbital and diethyl ether on the plasma glucose disappearance rate, IR, IRS-1 and IRS-2 tyrosine phosphorylation, IRSs association with PI 3-kinase, Akt and Erk phosphorylation, in liver and muscle of rats. Fasting plasma glucose levels were higher in animals anaesthetized with ether. No differences in plasma glucose disappearance rates were observed, however. Insulin-induced IR, IRS-1 and IRS-2 tyrosine phosphorylation, association of these substrates with PI 3-kinase and Akt and ERK phosphorylation were similar in the three groups of animals in both tissues. These data suggest that both thiopental and pentobarbital may be used in studies where changes in insulin signaling are being measured and where adequate general anaesthesia is required. (c) 2005 Elsevier Inc. All rights reserved.762022872297