Molecular cloning and characterization of thyroid hormone receptors in teleost fish

We describe an efficient reverse transcription/PCR procedure that allows the isolation of large fragments corresponding to TR and TR of a wide range of teleost fish

O Marchand; R Safi; H Escriva; E Van Rompaey; P Prunet; V Laudet

2004

Scholarcy highlights

  • Thyroid hormones play an important role in the growth, development and metabolism of vertebrates
  • reverse transcription/PCR reactions were performed with RNA from a wide variety of teleost fish that represent most of the diversity of this very large clade: an anguilliform, the eel; two salmonids, the rainbow trout and the Atlantic salmon; a pleuronectiform, the turbot; a tetraodontiform, the fugu; a cypriniform, the zebrafish; and a perciform, the tilapia
  • RT-PCR isolation of TRs from a wide variety of teleost fish In order to isolate specific probes for TR or TR from a wide variety of fish, we designed a series of degenerated PCR primers that can be used after reverse transcription
  • For each fish RT-PCR, fragments were obtained at the expected size for TR or TR
  • Cloning and phylogenetic analysis of teleost TRs In this study, we have set up an RT-PCR stategy that allows the isolation of large probes corresponding to the TH receptors and in a wide variety of teleost fish
  • The choice of species used, i.e. basal teleosts such as the eel as well as members of a derived order of euteleostei such as flatfish, makes us confident that, using the same primers, it will be possible to isolate the TRs of any teleost
  • Our transactivation experiments suggest that the Salmo TR binds T3 with an affinity comparable to that of amphibian or mammalian receptors, since we observed a convincing dose/response curve, with a minimal transactivation for 1 nM T3, which is in accordance with the Kd for other TRs

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