Accumulation of mRNAs encoding H1 zero and H3.3, two histone replacement variants, was studied in differentiating cortical neurons, cultured in a serum-free medium, with or without triiodothyronine (T3) supplementation. We found that the levels of both H1 (zero) and H3.3B mRNAs decrease in isolated neurons between the 2nd and 5th day of culture to the same extent as in vivo. At the same time, an active synthesis of the corresponding proteins was evidenced. The effects of transcription inhibition by actinomycin D and the results of nuclear run-on experiments suggest that H1 zero and H3.3 expression is regulated mainly at the posttranscriptional level. Concerning T3, only marginal effects were noticed, apart from up-regulation of both histone mRNAs at 2 days in culture. We propose one model for posttranscriptional regulation of the analyzed genes and discuss potential relationships to remodelling of chromatin.