A cloned thymosin beta-10 cDNA was used to study modulation of thymosin beta-10 mRNA levels in the rat B104 neuroblastoma cell line in response to retinoic acid. Northern blot analysis revealed the presence of a single greater than 600-nucleotide thymosin beta-10 mRNA species that was constitutively expressed in proliferating neuroblastoma cells. Addition of retinoic acid to the culture medium induced a dose- and time-dependent increase in thymosin beta-10 mRNA abundance. Additional studies showed that although thymosin beta-4 and beta-10 are coexpressed in this cell line, the stimulatory action of retinoic acid is specific for the thymosin beta-10 gene. Serum was found to augment the stimulatory action of retinoic acid. Blockade of protein synthesis with cycloheximide abrogated the stimulatory action of retinoic acid upon thymosin beta-10 mRNA accumulation; this observation suggests that activation of the thymosin beta-10 gene in this cell line by retinoic acid is dependent upon the de novo synthesis of a labile protein. Collectively, these findings demonstrate that the developmentally regulated thymosin beta-10 gene is a target for morphogenic retinoids in cells derived from the neural crest.