Embryonic lethal effect of expressing a dominant negative mutant human thyroid hormone receptor alpha1 in mice.

Abstract

Resistance to thyroid hormone (RTH) is caused mainly by mutations of the thyroid hormone receptor (TR) beta gene. Although, in vitro, TRalpha1 and TRbeta1 mutants exhibit similar dominant negative effects against wild-type TR, no TRalpha mutants have ever been identified in RTH patients. It has been postulated that mutations in TRalpha gene may be lethal, compensated completely by intact TRbeta or associated with phenotypic manifestations different from RTH. To investigate the consequences of mutant TRalpha1 expression in vivo, we tried to generate two different lines of transgenic mice which express a strong or a weak dominant negative mutant TR alpha1, respectively. First, we expressed betaF451X identified in a patient with severe RTH and alphaF397X, which has an identical C-terminal truncation and a similarly strong dominant negative potency to betaF451X, under the control of human polypeptide chain elongation factor 1alpha promoter. Six betaF451X-transgenic mice were born from 223 transferred embryos, giving a transgenic frequency of 2.7%. By contrast, expression of alphaF397X resulted in quite a low transgenic frequency with 0.39% of the transferred embryos bearing the transgene. Only three transgenic mice were born with no apparently overt abnormalities, of which one male produced F1 offspring. The transgenic progeny expressed alphaF397X in the testis but we did not succeed in generating transgenic mice expressing alphaF397X in multiple organs. To avoid toxic effects mediated by a strong dominant negative activity of mutant TRalpha1, we exchanged alphaF397X for alphaK389E, which has an identical missense mutation and a relatively weak transdominant potency as betaK443E identified in a patient with mild RTH. When expressed by cytomegalovirus immediate early enhancer-chicken beta-actin promoter, we did not succeed in creating alphaK389E-transgenic mice despite three independent transgene-injections. These findings define crucial in vivo functions of mutant TRalpha1s during mouse fetal development and suggest the possibility that the expression of a dominant negative mutant TRalpha1 in extensive tissues from the early embryonal stages might be lethal.