Cold exposure provides a reproducible model of improved glucose turnover accompanied by reduced steady state and glucose-induced insulin levels. In the present report we performed immunoprecipitation and immunoblot studies to evaluate the initial and intermediate steps of the insulin-signaling pathway in white and brown adipose tissues, liver and skeletal muscle of rats exposed to cold. Basal and glucose-induced insulin secretion were significantly impaired, while glucose clearance rates during glucose tolerance test, and the constant for glucose decay during a 15-min insulin tolerance test were increased, indicating a significantly improved glucose turnover and insulin sensitivity in rats exposed to cold. Evaluation of protein levels and insulin- induced tyrosine (insulin receptor, insulin receptor substrates -1 and -2, ERK) or serine (Akt) phosphorylation of proteins of the insulin signaling cascade revealed a tissue-specific pattern of regulation of the molecular events triggered by insulin in such a way that in white adipose tissue and skeletal muscle an impaired molecular response to insulin was detected, while in brown adipose tissue an enhanced response to insulin was evident. In muscle, white and brown adipose tissues, increased 2D-glucose uptake was detected. Thus, during cold exposure there is a tissue-specific regulation of the insulin-signaling pathway, which seems to favor heat-producing brown adipose tissue. Nevertheless, muscle and white adipose tissue are able to uptake high amounts of glucose even in face of an apparent molecular resistance to insulin.