Nuclear receptors for L-triiodothyronine in trout erythrocytes.
We have developed an in vitro assay to evaluate saturable specific binding of triiodothyronine (T3) by erythrocyte (RBC) nuclei isolated from rainbow trout. Our results indicate that the nuclei contain a T3-saturable protein which binds T3 with temperature, and pH dependency, high T3 affinity (Ka = 1.6 X 10(9) M-1), and relative thyroid hormone (TH) analog affinities (TRIAC greater than T3 greater than T4 greater than rT3 greater than T2) which are characteristic of TH receptors in other vertebrates. Our estimate of the maximal T3 binding capacity (MBC) of nuclei isolated from heterogeneous populations of RBCs at different maturational stages (MBC = 3.6 fM/mg DNA; 13 sites/nucleus) was 10-100 times lower than would be expected of a TH-responsive tissue. Differential cell counts revealed that 1% of the RBCs in our trout were immature (pro-RBCs). Pro-RBCs, in contrast to mature RBCs, contain abundant heterochromatin, mitochondria, and polyribosomes, and synthesize hemoglobin. Evaluation of binding data for RBC nuclei taken from trout in which erythropoiesis was stimulated by prior bleeding indicated that MBC was directly proportional to the absolute number of pro-RBC nuclei in the incubation. Our maximum estimate of MBC for pro-RBC nuclei (458 fmol/mg DNA; 1781 sites/nucleus) falls within the range of MBC values reported for other vertebrate TH-responsive tissues. These data indicate that RBCs of rainbow trout contain a nuclear protein (putative receptor) which binds T3 with characteristics similar to the TH receptor of higher vertebrates, and that nuclear T3 binding may be diminished during RBC maturation.
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D004920:Erythropoiesis; D005260:Female; D007700:Kinetics; D008297:Male; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D012487:Salmonidae / Q000097:blood*; D014284:Triiodothyronine / Q000097:blood*; D014337:Trout / Q000097:blood*
D011988:Receptors, Thyroid Hormone; D014284:Triiodothyronine