Thyroid hormone (T3) has previously been shown to regulate visual function in experimental animals and humans. To determine if T3 exerts direct effects on retinal function, cultured human fetal retinal pigment epithelial (RPE) cells were tested for the presence of thyroid hormone receptors (TRs) and T3 responses. Using TR-isoform-specific reverse-transcriptase polymerase chain reaction techniques, mRNA was detected for α1, α2 and β1 TR isoforms. Immunohistochemistry using a polyclonal antibody that simultaneously recognizes α1, α2 and β1 TRs showed nuclear staining of the fetal RPE. Specific binding of 125I-T3 to RPE cell nuclear extracts was detected, and Scatchard analysis revealed a Kd of 110 pM. To determine if RPE cells can respond to T3, hyaluronic acid (HA) levels in cell culture media were measured after 2, 4 or 6 days of growth in medium containing 10–7 M T3. T3 inhibited accumulation of HA in the cell culture medium of RPE cells. This effect was not evident at 2 days, but at 4 days there was 42.8% less HA in cell culture medium of RPE cells grown in 10–7 M T3 (p < 0.01, t test). The effect persisted through 6 days, when there was 46.3% less HA in cell culture medium of RPE cells grown in 10–7 M T3 (p < 0.001, t test). The data indicate that human fetal RPE cells are a direct target for thyroid hormones.