Immunocytochemical localization of aminopeptidase N on the cell surface of isolated porcine thyroid follicles.
The ultrastructural location of aminopeptidase N on the cell surface of isolated porcine thyroid follicle cells was studied with immunocytochemistry using antibodies against intestinal aminopeptidase N and protein A-colloidal gold. Gold particles, indicating immunoreactivity, were selectively attached to the apical cell surface. Occasionally, there was a sparse labelling of the basal cell surface. In follicles kept at 4 degrees C most gold particles at the apical cell surface appeared as clusters, with each gold particle situated at a constant distance of about 20 nm from the membrane surface. The gold particles were concentrated on the membranes of microvilli, in comparison to the smooth (intermicrovillar) portions of the apical plasma membrane. In follicles incubated at 37 degrees C for 5-180 min gold particles were slowly internalized by predominantly smooth-surfaced micropinocytic vesicles and subsequently appeared in colloid droplets and lysosomes. Gold particles were not observed in Golgi cisternae. TSH did not appear to influence the rate of internalization. TSH-induced pseudopods were unlabeled. Our electron-microscopic observations confirm previous immunofluorescence-microscopic evidence that aminopeptidase N is selectively expressed in the apical plasma membrane domain in the thyroid follicle cell. Furthermore, aminopeptidase N appears to be distributed in microdomains within the apical plasma membrane. Earlier indications of molecular differences between the pseudopod membrane and the apical plasma membrane proper are further emphasized.
D000626:Aminopeptidases / Q000378:metabolism*; D000818:Animals; D018826:CD13 Antigens; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D002478:Cells, Cultured; D008854:Microscopy, Electron; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D013552:Swine; D013961:Thyroid Gland / Q000166:cytology / Q000201:enzymology* / Q000648:ultrastructure
D000626:Aminopeptidases; D018826:CD13 Antigens