Cellular location of cytosolic triiodothyronine binding protein in primary cultures of fetal rat brain.

The evolution of a cytosolic triiodothyronine (T3) binding protein was studied in primary cultures of fetal rat brain. These cultures exhibited neuronal characteristics during the first week. T3 binding activity in cell supernatants increased during this period from 39 +/- 7 (mean +/- SD) to 159 +/- 24 fmoles T3/culture flask. A similar increase was observed in the soluble proteins. After day 8, neuronal death occurred and glial cells multiplied and differentiated. On day 11 an 86% drop in the binding activity was observed (24 +/- 7 fmoles T3/culture flask); the pool of soluble proteins remained stable. Scatchard analysis revealed two types of binding site in both 7- and 14-day cultured cell cytosols. Binding affinities were similar in both cytosols (KA1 approximately 1.5 X 10(9) M-1, KA2 approximately 1 X 10(8) M-1); in contrast, the number of sites was 4-fold smaller in 14-day cytosols. In subcultures mostly composed of glial cells, almost the same affinities were measured, but the numbers of both types of sites were 20 times smaller than in 7-day cells. These results show that in cell cultures from embryonic rat telencephalon, cytosolic T3 binding protein is mainly located in the neurons.