Organotypic cultures of locust embryo central nervous system (CNS) were used to study the influence of hormonal factors on neurite outgrowth. Explants from the third thoracic ganglion (embyronic day 9) were grown in a serum-free medium and exposed to insulin, neuroparsin, a recently characterized insect neurohormone, somatostatin and two insect hormones: 20-hydroxyecdysone and juvenile hormone. These hormonal factors were tested either alone or in several combinations. In our culture system both insulin and neuroparsin promoted extensive neurite outgrowth. Their effects were quantitatively similar. Somatostatin had no effect on neurite formation, but was a growth factor for glial cells. 20-hydroxyecdysone was a required factor for glial cell and neurite survival. Juvenile hormone did not have any observable effects in this culture system. When insulin was added in combination with 20-hydroxyecdysone the stimulation of neurite outgrowth exceeded that seen when insulin was added alone. A similar enhancement was observed with the combination of neuroparsin and 20-hydroxyecdysone. We conclude that in locusts: (i) insulin can be implicated as a neurotrophic factor as in vertebrates; and (ii) neuroparsin, a locust neurohormone, can also be characterized as a neurotrophic factor for the locust. Moreover, both neurohormones interact synergistically with the steroid hormone 20-hydroxyecdysone to enhance neurite growth.