Intestinal sucrase activity, absent in the suckling rat, is evoked precociously by thyroid hormones. Since enterocytes migrate from crypt to villus tip, the question arises: at which level of the villus--crypt columns do the enterocytes respond with an increase of sucrase activity to thyroxin (T4)? Suckling rats (11 days old) were injected daily, subcutaneously, with T4 (2 micrograms/g BW/day). They were sacrificed 1, 2, and 3 days later. Sucrase activity was determined in homogenates of the entire jejunal wall and in serial homogenates of the villus-crypt columns using cryostat sectioning. Maximal increase of sucrase activity was seen after 3 days in the lower villus. Experiments were also done using rats adrenalectomized on day 10 to exclude the effect via precocious maturation of adrenal cortex. The same results were obtained. In conclusion, T4-evoked increase of sucrase activity occurs first in enterocytes that are nearest the villus-crypt junction.