BACKGROUND Few attempts have been made to clarify the relational development of the hypothalamo-adenohypophysial and -neurohypophysial systems in species higher than amphibians. METHODS The appearance and topographical distribution of endocrine and neuroendocrine cells and fibers in these systems were immunohistochemically examined in the larvae of Xenopus laevis from immediately before hatching (stage 32, Nieuwkoop and Faber's classification) to the end of metamorphosis (stage 66). RESULTS (1) Each endocrine cell differentiated until the middle premetamorphic period. MSH cells initially appeared in the posterior half of the pituitary anlage at stage 35/36, followed by the differentiation of GH cells at stage 39 in the middle part, PRL cells at stage 46 in the anterior half of the pituitary anlage, and LH cells at stage 50 in the posterior two thirds of the pars distalis. With the progression of development, the cells which differentiated at early stages shifted from their initial positions; MSH cells, to the pars intermedia; and GH cells, to the posterior half of the pars distalis. 2) Oxytocin and vasopressin fibers were observed at stage 47/48 in the median eminence, and converged to the pars nervosa at later stages. 3) Neuroendocrine fibers innervated the median eminence during the middle premetamorphic to prometamorphic period: SOM fibers, at stage 45; CRH, 47/48; GRH, 48; dopamine, 58; and LHRH, 60. The cells containing these hormones were observed in the (presumptive) preoptic and/or infundibular nuclei. CONCLUSIONS These results suggest the following three chronological steps in the development of hypothalamo-hypophysial systems and their target organs: independent development of target organs at early developmental stages; appearance of hypophysial hormones to control the development of target organs at middle developmental stages; appearance of hypothalamic hormones to control the function or maturation of the hypophysis at late developmental stages.