We have shown in previous work that long-term high-protein diet treatment (420 days) induces important biochemical and stereological changes in rat liver mitochondria. In this paper we have studied the time course for these changes in rats fed a high-protein diet for 30, 90, 180, and 420 days. The liver carbamoyl-phosphate synthetase I (ammonia), which represents 15-20% of the mitochondrial protein, increased ca. 2.5-fold in 30 days, with no further significant changes during the treatment. This increase was accompanied by an increment in the serum urea levels and a diminution in the half-life of blood urea, which could be interpreted as compensatory mechanisms for detoxification of blood and for maintaining osmotic pressure. The stereological study indicates that there is an enlargement of individual mitochondria in rats fed the high-protein diet, and that the maximum enlargement occurred at 90 days of treatment. The analysis of data shows, however, that the increase in mitochondrial volume density was due mainly to proliferation of normal mitochondria. These mitochondria were functionally normal as demonstrated by the unaltered P:O ratio during treatment. The total content of liver amino acids was increased, and the taurine/glycine ratio (which has been related to metabolic stress) was greatly increased. The possible correlation between the increases of both liver taurine levels and cell volume is discussed.