5-Hydroxytryptamine (5-HT)-like immunoreactive neurones were studied during the development of myenteric plexus explant cultures from the guinea-pig taenia coli over a period of 3 weeks in vitro, using immunofluorescence histochemistry. Brightly fluorescent 5-HT-like immunoreactive neuronal cell bodies and fibres were found in all ages of cultures examined. In mature cultures, where the histotypic organization resembles that of the plexus in vivo, the pattern of immunoreactivity was strikingly similar to that previously described for in situ preparations. These culture preparations may therefore be useful models for the study of the development of putative serotonergic neurones. High-performance liquid chromatography was used to measure 5-HT levels in fetal calf serum, a supplement of the culture medium used in this study. 5-HT levels of 0.48 X 10(-5) M to 1.74 X 10(-4) M were found in 4 batches of this serum, indicating that some of the immunoreactive neurones observed in the cultures may have selectively taken up 5-HT during development in vitro.