Measurement of plasma thyroxine binding protein in relation to thyroidal condition in the turtle, Trachemys scripta, by radioimmunoassay

Abstract

Polyclonal (rabbit) antisera were generated against a high-affinity plasma thyroxine (T 4) binding protein (TBP) purified from the turtle, Trachemys scripta, and used to develop a specific radioimmunoassay (RIA). The RIA demonstrated the presence of an immunochemically related protein in the plasma of several other species of Trachemys and in members of several other genera from the same family, Emydidae. Plasma from all nonemydids and some emydid genera either showed no competition or nonparallelism in RIA. The presence and level of radioimmunoassayable TBP in diverse species correlated with results of previous comparative measurements of T 4 binding activity. However, an immunoreactive protein of the same molecular weight as TBP was identified in all turtles by Western blot analysis. More detailed studies in T. scripta demonstrated that variations in plasma T 4 binding activity induced by experimental or environmental manipulations were related to differences in TBP concentrations. The concentration of TBP varied by orders of magnitude (from <1 to ca. 150 mg/liter) in euthyroid animals; levels showed ontogenetic changes (virtually absent in hatchlings) and were directly related to thyroidal status. Experimentally induced hypothyroidism (goitrogen treatment or surgical thyroidectomy) resulted in a marked suppression of TBP, and T 4 treatment prevented its decline or reinstated it. Thus, in the turtle, this T 4 transport protein may exist in higher concentrations and its levels are more variable and show a different relationship to thyroid activity than the analogous T 4 binding globulin (TBG) in mammals.