Investigations on myelinogenesis in vitro: developmental expression of myelin basic protein mRNA and its regulation by thyroid hormone in primary cerebral cell cultures from embryonic mice.

The concentration of myelin basic protein (MBP) mRNA in primary cultures of cells dissociated from embryonic mouse cerebra and grown in the presence of varying amounts of thyroid hormone was measured using a 32P-labeled cDNA probe and a dot-blot procedure. The cDNA probe contained 1.85 kilobases of the gene for MBP. The concentration of mRNA specific for MBP in control cells grown on a medium containing normal (euthyroid) calf serum increased with increasing age of culture. The greatest increase occurred between 15 and 35 days in culture (5.25-fold increase); whereas between 35 and 50 days in culture, the rate of accumulation slowed to yield a net increase of MBP mRNA of only 10%. The quantity of MBP mRNA was drastically diminished at all ages studied when the cells were grown from the sixth day onward on a medium containing hypothyroid calf serum. Although the amount of MBP mRNA in hypothyroid-treated cells did increase, the change in concentration was less (3.43-fold), and it peaked earlier (at 30 days). Unlike the euthyroid cells, after 30 days the MBP mRNA actually fell in the hypothyroid-treated cells. If hypothyroid media were supplemented with triiodothyronine (T3) on the eighth day in culture, the quantity of MBP mRNA in the cells was restored almost completely to the levels found in the control euthyroid cells at all ages. Therefore, the regulation of the synthesis of MBP by thyroid hormone is at least in part a pretranslational event; that is, thyroid hormone adjusts the concentration of the mRNA specific for MBP.