EFFECT OF LEAD ON TRH AND GRF BINDING IN RAT ANTERIOR-PITUITARY MEMBRANES

The effect of lead on binding of the hypothalamic peptides thyroid releasing hormone (TRH) and growth hormone releasing factor (GRF) to rat anterior pituitary receptors was examined in this study. Concentrations of lead ranging from 0.01 to 1-mu-M did not alter [H-3]TRH binding; concentrations above 1-mu-M increased TRH association with pituitary receptors. A previously uncharacterized ligand, [I-125]GRF (human 1-44 amide), was used to examine the binding of GRF to anterior pituitary receptors. A high affinity site (GRF(H) = 18.1%, K(H) = 11.5 pM) was displaced by human growth hormone releasing factor (hGRF) (1-44)-NH2 or hGRF (1-29)-NH2 but not by rat growth hormone releasing factor (rGRF) (1-29)-NH2. Use of this ligand also revealed a class of low affinity binding sites (GRF(L) = 81.9%, K(L) = 0.39-mu-M) which has not been previously described. The low affinity site could be displaced by hGRF (1-44)-NH2, hGRF (1-29)-NH2 and rGRF (1-29)-NH2. A synthetic growth hormone releasing peptide (GHRP) also interacted with the low affinity GRF binding site. Lead dose-dependently displaced the binding of [I-125]GRF to its pituitary receptors. The IC50 of lead for inhibiting [I-125]GRF binding was 0.195 mM added lead or 52 pM free lead. These data suggest that one mechanism by which lead may affect pituitary function is through inhibition of receptor binding.