It has been reported that oligodendrocytes do not contain nuclear T3 receptors, which is in apparent contradiction with the well-known effects of thyroid hormones on myelination. In this study we have reexamined the presence of receptors in this cell population, using pure rat oligodendrocyte cultures. T3 binding was also studied with the use of pure rat astrocytes as well as in mixed neuronal-glial cultures. The latter are mainly neuronal during the first days in culture and essentially glial thereafter. Binding studies carried out in intact cells demonstrated the presence of high affinity-low capacity binding sites for thyroid hormones in pure cultures of oligodendrocytes. The maximal binding capacity was 50-60 fmol/100 micrograms DNA and the dissociation constant (Kd) 0.13 nM. Pure rat astrocyte cultures also contained high affinity sites for thyroid hormones, although receptor concentrations was 2-3 times lower than in oligodendrocytes or neurons. This was confirmed in pure cultures of chick astrocytes and in neuronal-glial cultures during the astroglial period. The relative affinity of the receptor for thyroid hormone analogs was triiodothyroacetic acid = T3 greater than T4 greater than tetraiodothyroacetic acid in oligodendrocyte and astrocyte nuclei, and the sedimentation coefficient of the receptor was approximately 3.8S in both cell types. These results demonstrate that nuclear T3 receptors similar to those found in neurons and astrocytes are also present in oligodendrocytes. This suggests that the effects of thyroid hormones on myelination could result from a direct action of the hormone in the oligodendrocytes.