Treatment of reaggregate pituitary cell cultures of 14-day-old female rats with nerve growth factor (NGF) augmented the number of [3H]thymidine ([3H]T)-incorporating lactotrophs in a dose-dependent manner (0.03-3 nM). At least during short-term treatment NGF increased the total number of cells expressing prolactin (PRL) mRNA and enlarged the cytoplasmic area occupied by PRL mRNA but did not affect the number of cells and the cytoplasmic area containing PRL, suggesting that NGF recruits lactotrophs expressing PRL mRNA but not yet PRL. NGF also stimulated [3H]T incorporation in ACTH cells but not in somatotrophs, thyrotrophs and gonadotrophs. In addition, NGF augmented the total number of [3H]T-incorporating cells to a much higher extent than was expected from its effect on lactotrophs and ACTH cells, suggesting NGF also stimulates [3H]T-incorporation in non-hormone producing cells (progenitors or stem cells?). Around 40% of these [3H]T-incorporating cells in both control and NGF treated cultures showed immunoreactivity for the transcription factor Pit-1 in the nuclei, which is twice the percentage expected (18%) if these [3H]T-incorporating cells were the only known Pit-1 expressing cells in the pituitary i.e. lactotrophs, somatotrophs and thyrotrophs. The present data suggest that NGF has a mitogenic effect on several cell lineages in the pituitary: lactotrophs, corticotrophs and non-hormone-containing cells. The high proportion of mitotic non-hormone containing cells that express Pit-1 is consistent with the proposed role of Pit-1 in cell proliferation in the developing lactosomatotroph lineage.