The effects of diazepam on the development of explanted stage 4 chick embryos were investigated. Diazepam, at 10-120 micrograms/ml, preferentially inhibited closure of the neural tube. This effect was reversible. Concentrations of 150-200 micrograms/ml inhibited not only neural tube closure but also blastodermal expansion, somite formation, and heart development in 52% of the embryos. Concentrations above 200 micrograms/ml were highly embryotoxic. Electron microscopy of affected neuroepithelial cells revealed that 1) apical surfaces were much smoother than usual and 2) apical filament bundles, which are generally thought to provide motive forces for uplifting of neural folds, were not well organized and often lacked alternating dark and light areas along their length. These findings and the fact that changes in cell surface topography reflect the contractile activities of underlying filament bundles suggest that the observed "smoothing" effect on apical cell surfaces and neural tube closure defects are due, at least in part, to the impaired ability of these filament bundles to contract.