Developmental changes in the activities of cystathionine beta-synthase and cystathionine gamma-lyase were measured in six regions of rat brain. On day-1, no differences were observed in the activities of cystathionine beta-synthase and cystathionine gamma-lyase among these regions, the values being about 40 nmol/h/mg protein, and 3 nmol/h/mg protein, respectively. Cystathionine beta-synthase activity increased gradually during development at almost the same rate in each region, reaching the adult level at week-4 (about 4-fold). Cystathionine gamma-lyase activity also increased during development, reaching adult level at week-2. But, the increase of enzyme activity in the cerebellum (about 1.8-fold) was clearly lower than that in the other regions (about 4-fold). Cystathionine gamma-lyase content in the various regions of week-3 rat brain estimated by immunoblotting was consistent with the enzyme activity, and the enzyme level in the cerebellum was lower than that in the other regions. Cystathionine content of cerebellum in week-3 increased rapidly during development, and was about five-fold more than that on day-1. However, cystathionine content in the other regions did not change during development. These findings indicated that at least one reason of the high content of cystathionine in the 3 weeks rat cerebellum was due to the low level of cystathionine gamma-lyase.