The unique organization of the teleost pituitary, in which cells are grouped according to their characteristic hormone, makes this a suitable model for studying pituitary paracrine interactions. In a number of fish, including tilapia, there are variations in the circulating levels of the gonadotropins and GH, which are elevated during the reproductive season, suggesting interactions between the reproductive and growth axes. The aim of this study was to investigate paracrine interactions between the gonadotrophs and somatotrophs in the tilapia pituitary. Initially, dispersed pituitary cells were separated on a density gradient in which the gonadotrophs were found in the least dense fractions, and the somatotrophs were concentrated in the densest fraction. After 4 days in culture, cells in the least dense fractions showed characteristic cytoplasmic extensions not seen in the somatotrophs, which appeared small and failed to form aggregates; somatotrophs were found, however, attached to other non-GH cells. Staining of the nuclei with 4,6-diaminidino-2-phenyl-dihydrochloride revealed that the isolated somatotrophs had undergone nuclear condensation and fragmentation typical of apoptosis. Addition of either estradiol or human recombinant insulin-like growth factor I (IGF-I; 10 nM) to the somatotroph cultures increased the number of cell aggregates and reduced the number of condensed or fragmented nuclei. Immunocytochemical studies on pituitary sections revealed IGF-I immunoreactivity in regions of the proximal pars distalis that stain with gonadotropin IIbeta antisera and also in regions of the rostral pars distalis characteristic of corticotrophs; immunoreactive IGF-I was never seen in the region of the somatotrophs. Incubation of cells from the different fractions with testosterone (10 nM; 24 h) revealed that cells of the least dense fractions, which were rich in gonadotrophs, possessed aromatizing ability, which was absent in the somatotroph-enriched fraction. These results suggest that estradiol and IGF-I, both generated from cells other than the somatotrophs, may exert antiapoptotic effects and thus possibly control the size of this population of cells.