The sequential changes in the histological pattern of anterior pituitary cytodifferentiation of the rat are described. The first labeled cells were ACTH positive and were detected in the pars tuberalis on postconceptual day 13. On day 14 ACTH cells also appeared in the ventral periphery of the pars distalis. On fetal day 15 the pars tuberalis anlage was characterized by numerous well-stained ACTH cells and by some weakly labeled FSH-β, LH-β, TSH-β, GH and PRL cells while the pars distalis showed only ACTH positivity. On day 16 of gestation the ACTH cells were equally distributed throughout the whole pars distalis, while LH-β, FSH-β, TSH-β, PRL and GH immunoreactive cells were localized either in the ventral region of the pars distalis only or were evenly distributed throughout the pars distalis. The present immunocytochemical data suggest that in the pars distalis the hypophyseal cell differentiation follows a clear rostrocaudal, ventrodorsal direction and that the time sequence of the functional differentiation of the adenohypophysis is pars tuberalis, pars distalis and pars inter