Oral cholecystographic agents induce a decrease in serum T3 and an increase in serum T4 and rT3 concentrations in normal subjects. However, their effect on TSH secretion in man is unclear. In the present study, the serum TSH concentration was increased above baseline values 5 days after the administration of 3 g iopanoic acid (IA) in five of six euthyroid volunteers (an increase of borderline significance), and the TSH response to TRH was significantly augmented after IA administration. In four other euthyroid subjects who had received IA 3 days earlier, the administration of T3 (5 microgram, five times daily) for the next 2 days restored the serum T3 concentration toward baseline values and prevented the IA-induced increase in TRH-stimulated serum TSH concentrations. It is concluded from the present study that IA enhances pituitary thyrotroph sensitivity to TRH and that this effect may be related, at least in part, to the IA-induced decrease in circulating T3 as well as the previously demonstrated inhibitory effect of IA on the pituitary conversion of T4 to T3. This decrease in the serum T3 concentration after IA administration is due primarily to inhibition of peripheral 5-monodeiodination of T4 and, possibly, to a direct inhibitoy effect of excess iodide on the release of T3 from the thyroid.