The effects of aging on plasma concentration of thyroxine (T4) and cortisol and on responses of these hormones to low ambient temperatures were determined in the dog. Female beagle dogs were divided into three age groups: old, adult, and puppies. The mean (+/- SD) ages were 11.4 +/- 0.2 years, 3.0 +/- 0.4 years, and 7.6 +/- 0.2 weeks, respectively. All dogs came from a genetically homogeneous colony and were free from any disease. The adult and old dogs were used during anestrus. Based on four daily blood samples, the mean (+/- SE) T4 level in the old dogs (2.8 +/- 0.1 microgram/dl) was significantly (P less than 0.001) lower than that in the adults (4.2 +/- 0.2 micrograms/dl) and puppies (4.4 +/- 0.2 micrograms/dl). By contrast, mean plasma cortisol levels in the old dogs (21.1 +/- 3.1 ng/ml) and adults (15.4 +/- 2.4 ng/ml) were significantly higher than those in the puppies (7.2 +/- 1.1 ng/ml). No significant changes in plasma T4 and cortisol occurred in any of the three age groups at 22 degrees C or during exposure to 10 or 4 degrees C. Exposure to -5 degrees C, however, produced significant increases in T4 (greater than 130% by 5 hr) and cortisol (greater than 280% by 1 hr) in adult dogs. This temperature produced only a modest increase in T4 (70% by 3.5 hr) and no change in cortisol in the old dogs. The puppies showed no change in T4 and cortisol during exposure to -5 degrees C. The results demonstrate that with advancing age, plasma T4 and cortisol concentrations change in opposite directions, thus supporting the hypothesis of a negative relationship between these two hormones. These results also show that the responses of these hormones to the stress of cold decline during aging and are not yet developed in the very young.