Evidence for the onset of feedback regulation of cortisol in larval summer flounder.

We investigated the functional development of feedback regulation of cortisol levels during early development in a marine teleost, summer flounder, using a novel pharmaceutical approach. Larvae were immersed for 4h in the glucocorticoid agonist dexamethasone (20 microM) and/or the glucocorticoid-receptor antagonist RU486 (0.12 microM) at 1, 7, or 21 days after hatching. The hypothesis was that, if feedback regulation were operational, tissue cortisol concentrations would be suppressed by dexamethasone and stimulated by RU486. Whole-body cortisol content of 1-day-old larvae was significantly decreased from 0.32 ng/g body weight (mean) to 0.08-0.12 ng/g by immersion in dexamethasone, RU486, or both, perhaps due to displacement of cortisol from the yolk sac. There were no changes in cortisol content among treatment groups in 7-day-old larvae. The expectations of our hypothesis were met in 21-day-old larvae. Immersion in RU486 increased cortisol content from 0.29 ng/g (control) to 2.00 ng/g, whereas immersion in dexamethasone (with or without RU486) suppressed cortisol to 0.03-0.04 ng/g. The results indicate that a fully functional hypothalamic-pituitary-interrenal axis is established by 3 weeks after yolk-sac resorption, but before the onset of metamorphosis in summer flounder. This is the earliest detection of feedback regulation in a teleost fish.

Cortisol is necessary for seawater tolerance in larvae of a marine teleost the summer flounder.

Larval-stage summer flounder (Paralichthys dentatus) were immersed in the corticosteroid-receptor blocker RU486 to test the effects of cortisol deficiency on salinity tolerance. Premetamorphic larvae held at 10 (near isosmotic) or 30 (hyperosmotic) parts per thousand ( per thousand) seawater survived well over 5d in 0, 0.012, or 0.12 microM RU486. However, at concentrations of 1.2 or 3.6 microM RU486, mortality was significantly greater for larvae in 30 per thousand compared to larvae in 10 per thousand. In a separate experiment, the ability of RU486 to inhibit tolerance to hyperosmotic medium (30 per thousand) was confirmed; immersion at 1.2 microM RU486 induced mortality of larvae in the metamorphic climax stage held at 30 per thousand, but not 0 or 10 per thousand. Mortality due to RU486 in pre- or prometamorphic stage larvae was prevented by concurrent immersion in cortisol at concentrations approximately 10-200 times greater than RU486, indicating that the action of RU486 was specific to antagonism of cortisol. The efficacy of 1.2 microM RU486 in reducing survival in 30 per thousand was found to be stage-dependent and exhibited the following hierarchy for fastest time to 50% mortality: prometamorphosis>metamorphic climax>premetamorphosis. In a 5-d pretreatment of pre- or prometamorphic larvae by immersion in 20 microM cortisol and/or 0.12 microM RU486 at 30 per thousand, only RU486 had a limited effect on decreasing survival when larvae were challenged with abrupt exposure to 50 per thousand. In total, the results evidence for the first time a necessary role for cortisol in seawater tolerance of a larval marine teleost.