The role of thyroid hormones during the development of eye pigmentation in the Pacific bluefin tuna (Thunnus orientalis).

Thyroid hormones (THs) are essential for the embryonic and post-embryonic development of fish. We studied the role of THs during the early, post-embryonic, development of Pacific bluefin tuna. Embryos were treated with L-thyroxine (T(4)) or the anti-thyroid drug methimazole (MMI), and reared in microtitre plates for 3 days. Immersion in MMI, but not T(4), led to retardation of retinal pigment epithelium (RPE) pigmentation 3 days post-hatching (dph). Concurrent immersion in T(4) and MMI had no effect of RPE pigmentation. We also measured the expression of TRalphaA, TRalphaB, and TRbeta mRNA using real-time RT-PCR. Treatment with MMI significantly reduced TRbeta mRNA expression. Taken together these results suggest that the development of RPE pigmentation is mediated by TH, most likely via TRbeta.

Characterization of thyroid hormones and thyroid hormone receptors during the early development of Pacific bluefin tuna (Thunnus orientalis).

We studied the profiles of 3,5,3'-l-triiodothyronine (T3), thyroxine (T4), and thyroid hormone receptors (TRs) in Pacific bluefin tuna (Thunnus orientalis) during embryonic and post-embryonic development. Both T3 and T4 were detected in embryos just before hatching, and it was found that the levels of both were increased in postflexion fish. The thyroid follicles were increased in both size and number in postflexion fish compared with preflexion fish. A TRbeta cDNA clone was generated by RACE. Two TRalpha cDNA clones were also partially identified and analyzed by real-time RT-PCR in this study. The TR mRNA levels in embryos were determined, and these were found to be lower than those in preflexion fish. Therefore, we considered that thyroid hormones function during early post-embryonic development as well as during embryonic development. Moreover, there was a peak in the TR mRNA level during postflexion stages, as seen during metamorphosis in Japanese flounder and Japanese conger eel. It is possible that thyroid hormones control the early development of scombrid fish through TRs, as they do for Pluronectiformes and Anguilliformes.

Characterization of transthyretin in the Pacific bluefin tuna, Thunnus orientalis.

A cDNA encoding transthyretin was cloned from the Pacific bluefin tuna (Thunnus orientalis). This cDNA contains a complete open reading frame encoding 151 amino acid residues. The deduced amino acid sequence is 81% and 55% identical to the gilthead seabream and common carp forms, respectively, and 33-39% to mammalian, reptilian, and amphibian forms. A 1.0-kb transcript was found in the the liver and ovary; the liver is the main source of this protein. Analysis of triiodo-L-thyronine (T(3)) and L-thyroxine (T(4)) binding demonstrated that both T(3) and T(4) bind to bluefin transthyretin. The binding activity of T(3) for bluefin transthyretin is higher than that of T(4). These results indicate that bluefin transthyretin acts as a transporter of thyroid hormones (THs) in the plasma, and plays an important role in the function of THs in target cells.

The histological analysis, colorimetric evaluation, and chemical quantification of melanin content in 'suntanned' fish.

Human melanocytes respond to UV irradiation by increasing the synthesis of melanin. While much is now understood of the pathways governing this process and the nature of the melanin synthesized, little is known of melanins produced by lower vertebrates and their capacity to respond to UV. Here we report that a fish, red seabream, can undergo 'suntanning'. Histological, colorimetric and chemical assays were performed for suntanned red seabream fish bred in net cages to analyse the melanins and compared with shaded or wild red seabream fish. For color evaluation, the L* values of suntanned fish were dramatically lower than those in the other two groups. Pyrrole-2,3,5-tricarboxylic acid (PTCA), an indicator of eumelanin, was detected in suntanned fish at five times higher levels than in shaded or wild fish while 4-amino-3-hydroxyphenyl-alanine (4-AHP), a marker for pheomelanin, could not be detected in any of the samples. Histological analysis showed that melanocytes in the suntanned skin enlarged and increased in number to form a monolayer at the surface of the skin. Analysis of L* values and PTCA levels showed quite a high correlation coefficient (r = -0.843). When comparing shaded and wild red seabream fish, the scores were closer but some significant differences were still found in some body areas. These results indicate that eumelanin accumulates in suntanned fish during the increase in skin color, which is induced by sunlight, presumably by ultraviolet radiation.