Effect of melatonin on the response of the thyroid to thyrotropin stimulation in vitro.

Thyroidal-melatonin interactions are of particular importance to amphibian development since the thyroid controls the progress of metamorphosis while melatonin may coordinate its rate with prevailing environmental conditions. Melatonin antagonized thyroxine (T4) action at the tissue level and directly inhibited baseline T4 secretion in culture, so the present work sought to determine if it antagonized the response of the thyroid to thyroid stimulating hormone (TSH) as well. A preliminary experiment showed that, in Rana pipiens, the concentration of TSH (0.2 microg/ml) used in the culture of tadpole thyroids stimulated T4 secretion as much as frog pituitaries, but more than late premetamorphic tadpole pituitaries. There was no significant effect of 1 to 15 microg/ml melatonin in TSH-containing thyroid cultures of various Rana species of tadpoles and frogs in experiments with media collected once every 24 or 48 hr, although 15 microg/ml melatonin tended to depress T4 secretion. In a final experiment, a higher melatonin concentration was used as well as more frequent media collections. Thyroids from prometamorphic Rana catesbeiana tadpoles were cultured in L-15 media with periodic stimulation by 0.2 microg/ml TSH, or TSH and 10 or 100 microg/ml melatonin. Media were collected at the end of two 3-hr TSH pulses, and every 8 hr thereafter for the next 3 days. Melatonin was administered until the end of Day 2 while TSH was given only on Day 2 in addition to the original 3-hr pulses. The secretion of T4 was inhibited significantly by 10 microg/ml melatonin at only two of the early media collections. In contrast, 100 micro;g/ml melatonin significantly depressed T4 secretion in response to TSH at all but one interval and completely inhibited the thyroidal response to TSH reintroduced into the media on Day 2. The findings suggest that a high concentration of melatonin is inhibitory to the thyroidal response to TSH, but that lower concentrations do not significantly overcome the TSH stimulus.

Circadian rhythms of thyroid secretion, morphometry, and cell division in prometamorphic and climax Rana tadpoles.

Rhythmicity of thyroid follicle structure, cell division, and T4 secretion in vitro was studied in anuran tadpoles on a 12L:12D cycle with light onset at 0800 hr. Stage XIII Rana pipiens larvae had a significant circadian rhythm of follicle lumen diameter (acrophase 1052 hr). Follicle cell height was higher at 0900 hr than at other times but the rhythm was not significant. At Stage XX, both follicle cell height (acrophase 2318 hr) and cell division (acrophase 1929 hr) were rhythmic while lumen diameter, though highest at 0900 hr, showed no significant daily changes. Thyroids from Rana catesbeiana larvae at Stages XVII to XVIII had a significant circadian rhythm of T4 secretion (acrophase 2034 hr) in vitro as measured by radioimmunoassay of media collected every 6 hr for 24 hr. The findings indicate that the daily sequence of thyroid structure and function in preclimax Rana tadpoles includes larger follicle lumina early in the light and maximum cell division, T4 secretion, and follicle cell height within a 4-hr interval beginning around the onset of dark.