The relationship between ingested thyroid hormones, thyroid homeostasis and iodine metabolism in humans and teleost fish.

Oral l-thyroxine (T4) therapy is used to treat human hypothyroidism but T4 fed to teleost fish does not raise plasma thyroid hormone (TH) levels nor induce growth, even though oral 3,5,3'-triiodo-l-thyronine (T3) is effective. This suggests a major difference in TH metabolism between teleosts and humans, often used as a starting thyroid model for lower vertebrates. To gain further insight on the proximate (mechanistic) and ultimate (survival value) factors underlying this difference, the several steps in TH homeostasis from intestinal TH uptake to hypothalamic-hypophyseal regulation were compared between humans and teleosts, and following dietary TH challenges. A major proximate factor limiting trout T4 uptake is a potent constitutive thiol-inhibited intestinal complete T4 deiodination that is ineffective for T3. At the hepatic level, T4 deiodination, conjugation and extensive biliary excretion with negligible T4 enterohepatic recycling can further block teleost T4 uptake to plasma. Such protection of plasma T4 from dietary T4 may be particularly critical for piscivorous fish consuming thyroid tissue, rich in T4 but not T3. It would prevent disruption by unregulated ingested T4 of the characteristic acute and transient changes in teleost plasma T4 due to diel rhythms, food intake and stress-related factors. These marked natural short-term fluctuations in teleost plasma T4 levels are enabled by the relatively small and rapidly-cleared plasma T4 pool, stemming largely from properties of the plasma T4-binding proteins. Humans, however, due mainly to plasma T4-binding globulin, have a relatively massive circulating pool of T4 and an extremely well-buffered free T4 level, consistent with the major TH role in regulating basal metabolic rate. Furthermore, this large well-buffered and slowly-cleared plasma T4 pool, in conjuction with enterohepatic recycling and relaxation of hypothalamic-hypophyseal negative feedback, allows humans to temporarily 'store' ingested T4 in plasma, thereby sparing endogenous TH secretion and conserving thyroidal iodine reserves. Indeed, iodine conservation is likely the key ultimate factor determining the divergent evolution of the human and teleost systems. For humans, ingested iodine in the form of I-, or TH and their derivatives, is the sole iodine source and may be limiting in many environments. However, most freshwater teleosts, in addition to their ability to assimilate dietary I-, can derive sufficient I- from their copious gill irrigation, with no selective advantage in absorbing dietary T4 which would disrupt their natural acute and transient changes in plasma T4. Thus T4 may act also as a vitamin (vitamone) in humans but not in teleosts; in contrast, T3, naturally ingested at much lower levels, may act as a vitamone in both humans and teleosts.

A cDNA for a putative type III deiodinase in the trout (Oncorhynchus mykiss): influence of holding conditions and thyroid hormone treatment on its hepatic expression.

A putative rainbow trout type III deiodinase (D3) cDNA was amplified by PCR, using primers to evolutionarily conserved sequences. The RACE-derived complete cDNA was then identified by sequence comparison to that in tilapia and other vertebrates. The cDNA coded for a predicted 31,500 kDa protein of 278 amino acids, with a hydrophobic trans-membrane segment and with 80% similarity to tilapia D3 and 39% similarity to rainbow trout type II deiodinase (D2). It also showed a selenocysteine codon at position 141 and a putative SECIS element in the 3' untranslated end. In the liver, a second form of D3 was found that differed only at this 3' untranslated region; the coding region was identical in both forms. The D3 mRNA, measured by RT-PCR using primers located within the common, translated portion of the cDNA, was expressed in the brain and, depending on thyroidal status, in liver and kidney. Holding trout for 7 days in static water as opposed to flowing water caused increased plasma T4 levels, decreased hepatic D2 mRNA levels and T4 outer-ring deiodination (ORD) activity and increased D3 mRNA levels and T3 inner-ring (IRD) activity. Trout held in flowing water and fed T3 for 7 days showed increased plasma T3 levels and hepatic D3 mRNA levels and T3 IRD activity but decreased D2 mRNA levels and T4ORD activity. Trout held in static water and exposed to ambient T4 for 7 days showed increased plasma T4 levels and hepatic T3IRD activity but with no significant change in D2 or D3 mRNA levels. We conclude that hepatic D3 mRNA levels and T3IRD activity are enhanced and D2 mRNA levels and T4ORD activity are suppressed by adverse holding conditions or T3 treatment suggesting that the putative D3 cDNA and D2 cDNA represent respectively the genes determining T3IRD and T4ORD activities. However, there were changes in the ratios of mRNA levels to enzyme activity, raising the potential for post-transcriptional regulation and showing that mRNA levels alone may be unreliable indices of deiodinase activity. Post-transcriptional regulation of D3 enzyme activity may be influenced by the observed alternative 3' and 5' untranslated regions of the D3 mRNA.

Contaminant effects on the teleost fish thyroid.

Numerous environmentally relevant chemicals, including polychlorinated hydrocarbons, polycyclic aromatic hydrocarbons, organochlorine pesticides, chlorinated paraffins, organophosphorous pesticides, carbamate pesticides, cyanide compounds, methyl bromide, phenols, ammonia, metals, acid loads, sex steroids, and pharmaceuticals, exert acute or chronic effects on the thyroid cascade in the approximately 40 teleost fish species tested to date. Thyroid endpoints, therefore, serve as biomarkers of exposure to environmental pollutants. However, the mechanisms underlying thyroid changes and their physiological consequences are poorly understood because the thyroid cascade may respond indirectly and it has considerable capacity to compensate for abuses that otherwise would disrupt thyroid hormone homeostasis. Indeed, a xenobiotic-induced change in fish thyroid function has yet to be conclusively causally linked to decreased fitness or survival. Other complications in interpretation arise from the diversity of test conditions employed and the often indiscriminate use of numerous thyroid endpoints. Future work should be directed toward standardizing test conditions and thyroid endpoints and investigating causal links between thyroid changes and fish growth, reproduction, and development. Development may be particularly susceptible to thyroid disruption, and thyroid endpoints appropriate for early life stages need to be applied.

Thyroid of lake sturgeon, Acipenser fulvescens. I. Hormone levels in blood and tissues.

The authors measured thyroid hormone (TH) levels in plasma, whole carcass, and tissues of cultured 2-year-old immature lake sturgeon held in fresh water and in serum of adults at spawning time from the Winnipeg River. Circulating thyroxine (T4) and 3,5,3'-triiodothyronine (T3) levels were low (T4 approximately 0.3 ng/ml, T3 approximately 0.2 ng/ml) in all cultured fish and most adults, but a few of the latter had exceptionally high T4 and T3 levels. The percentages of blood TH found in erythrocytes were 19.5% (T4), 6.1% (T3) and 6.9% (reverse T3 = rT3). Equilibrium dialysis showed much higher percentages of plasma free (F) FT4 (1.1%), FT3 (0.4%), and FrT3 (3,3',5'-triiodothyronine = rT3, 3.0%) for sturgeon than for rainbow trout, indicating more limited TH binding to sturgeon plasma sites. However, concentrations of FT4 and FT3 were close to those reported for salmonids. T3 levels exceeded T4 levels in most extrathyroidal tissues of cultured sturgeon but in most cases were less than 0.1 ng/g and 10 to 100 times lower than reported for salmonids; only the whole brain T3 concentration (5.6 ng/g) approached that of salmonids. The digested thyroid contained 21.3 ng T3/g and 2.4 ng T4/g. The authors conclude that lake sturgeon have a low circulating reserve of bound TH but have FT4 and FT3 concentrations close to those of salmonids. The high thyroidal T3:T4 ratio and low tissue T4 levels suggest that, in contrast to teleosts studied to date, the thyroid may be a significant direct source of T3, the primary TH in sturgeon tissues. High serum T4 and T3 levels in some sturgeon at spawning time may suggest a thyroid role in reproduction.

Thyroid of lake sturgeon, Acipenser fulvescens. II. Deiodination properties, distribution, and effects of diet, growth, and a T3 challenge.

The authors studied the properties and tissue distribution of thyroid hormone (TH) deiodination activities measured in vitro at subnanomolar substrate levels for cultured 2-year-old lake sturgeon held at 12 to 15 degrees. We also studied the deiodination responses to an exogenous 3,5,3'-triiodothyronine (T3) challenge and to a diet-induced growth suppression. Thyroxine (T4) outer-ring deiodination (T4ORD), T4 inner-ring deiodination (T4IRD), T3IRD, and 3,3',5'-triiodothyronine (rT3)ORD activities were evident in liver and intestine. Their properties resembled those of teleosts. T3IRD and T4IRD activities predominated in brain. Low or negligible deiodination in any form occurred in gill, skeletal muscle, kidney, notochord, or immature gonad. Only T4ORD activity was evident in the thyroid, suggesting that it secretes some T3. T3ORD and rT3IRD activities were undetectable in any tissues. Hepatic T4ORD activity varied during the photophase and was highest during late morning. A dietary T3 challenge that doubled plasma T3 levels decreased hepatic T4ORD activity without altering any other deiodination pathways in liver, intestine, or brain. A diet change from trout pellets to ocean zooplankton reduced somatic growth and plasma T3 levels and increased hepatic and intestinal T3IRD activities and hepatic rT3ORD activity but did not alter hepatic or intestinal T4ORD activity. The authors conclude that plasma T3 in lake sturgeon can be derived both from the thyroid and from hepatic (and intestinal) T4ORD activity, which varies with sampling time and downregulates in response to a T3 challenge. However, a reduction in plasma T3 accompanying a change in diet and reduced growth was not due to a decrease in T4ORD activity; rather, it was due to an increase in hepatic and intestinal T3IRD activities. These results suggest a difference in emphasis in thyroidal regulation between sturgeon and certain teleosts.