Uptake, metabolism and subcellular distribution of [125I]T4 in calf thyroid slices

Previous work from our laboratory has shown that iodothyronines have a direct inhibitory action on the thyroid. In the present studies the uptake, metabolism and subcellular distribution of labelled thyroxine were analyzed. The entry of this hormone reaches a plateau after 15-20 min incubation and is temperature-dependent. The T/M values for T4 were much lower than those of labelled T3 and the curve was different from that obtained with [125I]. The influence of a series of compounds on the T/M values for thyroxine was studied. KI, T3, IOP, PTU and MMI were without effect. Absence of Ca++ in the buffer, or addition of KClO4 or ouabain caused a slight decrease, while TSH produced a stimulation in the T/M ratio. Calf thyroid slices dehalogenated thyroxine, producing both T3 and rT3. TSH increased the generation of these two compounds. Neither PTU nor IOP altered the production of T3 and rT3 significantly. The lack of effect of IOP on T3 and rT3 generation was confirmed in calf thyroid homogenates. However, and in agreement with previous reports, IOP significantly decreased production of both triiodothyronines in rat liver slices and in rat liver homogenates. When the subcellular distribution of labelled thyroxine was examined most of the radioactivity appeared in the soluble fraction and less than 1% was present in purified nuclei. The addition of unlabelled thyroxine to the slices only caused a significant displacement in the radioactivity of purified nuclei. The presence of labelled thyroxine in the nuclei was confirmed by paper chromatography.