Immunoelectron microscopic studies on the cell surface location of the thyroid microsomal antigen.

ABSTRACT

The cell surface location of the thyroid microsomal antigen was studied by immunoelectron microscopy. Isolated, open human thyroid follicles were incubated with patient sera containing high titers of microsomal autoantibodies. Cell surface-bound antibodies were detected by the immunogold technique using IgG-coated colloidal gold particles (10 or 15 nm). Immunocytochemical incubations were performed at 4 degrees C. Gold particles were concentrated at the apical cell surface of the follicle cells, while the basolateral cell surface was almost completely unlabelled. Quantitative evaluation of four experiments with follicle cells prepared from different patients showed that about 90% of the gold particles at the apical cell surface was associated with microvilli and that the concentration of gold particles at the microvillus membrane was, although with great intercellular variation, several times higher than that at smooth portions of the apical plasma membrane. This suggests that the microsomal antigen is organized in microdomains in the apical plasma membrane. In follicles labelled immunocytochemically at 4 degrees C and then incubated at 37 degrees C, gold particles were slowly internalized. The particles appeared in smooth and coated pits of the apical plasma membrane as well as in vesicles, vacuoles and lysosomes in the apical part of the cytoplasm. Membranes of TSH-induced pseudopods were always unlabelled. Our observations indicate that thyroid microsomal antigen immunoreactivity is present in the apical but not in the basolateral plasma membrane. The antigen with bound antibodies is internalized by micropinocytosis but not by macropinocytosis. The selective location of bound microsomal antibodies at the apical plasma membrane and their absence from the membrane of TSH-induced pseudopods are compatible with the idea that the microsomal antigen and thyroperoxidase are identical.