Quantification of MSH Receptors on Mouse Melanoma Tissue by Receptor Autoradiography

Abstract

MSH receptors on mouse melanoma tissue sections were quantified by receptor autoradiography, yielding results which were very similar to those obtained by a conventional receptor binding assay with isolated cells. In order to minimize nonspecific binding, it proved to be crucial to use a radioactive monoiodinated MSH radioligand retaining full biological activity and to apply the binding conditions developed for isolated cells to the incubation of whole tissue sections. The displacement curves obtained after quantitative analysis of autoradiograms from tissue sections yielded a K_D-value of the same order of magnitude (0.58 nM; average of n=7 experiments) as those obtained in the normal binding assay with isolated cells (1.2 nM; average of n=10 experiments). Similarly, receptor numbers per cell on tissue sections (14,700; n=7) did not differ markedly from those determined with isolated cells (10,500; n=10). These results demonstrate that receptor autoradiography can be applied to the quantification of peptide hormone receptors on peripheral tissues.