Summary
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° 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° 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 unlabelled.
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.
Similar content being viewed by others
References
Antonov VK, Vorotyntseva TI, Bessmertnaya LY, Mikhailova AG, Zilberman MI (1984) Role of intestinal brush border membrane aminopeptidase N in dipeptide transport. FEBS Lett 171:227–232
Barriere H, Chambard M, Muranyi-Kovacs I, Mauchamp J, Louvard D, Gabrion J (1986) Pseudopod membrane in TSH-stimulated thyroid cells: a specialized domain in the neighboring apical plasma membrane. Cell Tissue Res 245:159–170
Björkman U, Ekholm R, Denef J-F (1981) Cytochemical localization of hydrogen peroxide in isolated thyroid follicles. J Ultrastruct Res 74:105–115
Colas B, Maroux S (1980) Simultaneous isolation of brush border and basolateral membrane from rabbit enterocytes. Presence of brush border hydrolases in the basolateral membrane. Biochim Biophys Acta 600:406–420
Consiglio E, Salvatore G, Rall JE, Kohn LD (1979) Thyroglobulin interactions with thyroid plasma membranes. The existence of specific receptors and their potential role. J Biol Chem 254:5065–5076
Denef J-F, Ekholm R (1980) Membrane labeling with cationized ferritin in isolated thyroid follicles. J Ultrastruc Res 71:203–221
Denef J-F, Björkman U, Ekholm R (1980) Structural and functional characteristics of isolated thyroid follicles. J Ultrastruct Res 71:185–202
Eagle H (1959) Amino acid metabolism in mammalian cell cultures. Science 130:432–437
Ekholm R (1981) Iodination of thyroglobulin — an intracellular or extracellular process? Mol Cell Endocrinol 24:141–163
Ericson LE (1981) Exocytosis and endocytosis in the thyroid follicle cell. Mol Cell Endocrinol 22:1–24
Ferraci H, Bernadac A, Hovsepian S, Fayet G, Maroux S (1981) Aminopeptidase N is a marker for the apical pole of porcine thyroid epithelial cells in vivo and in culture. Cell Tissue Res 221:137–146
Ferraci H, Bernadac A, Gorvel JP, Maroux S (1982) Localization by immunofluorescence and histochemical labeling of aminopeptidase N in relation to its biosynthesis in rabbit and pig enterocytes. Gastroenterology 82:317–324
Gerard C, Gabrion J, Verrier B, Reggio H, Mauchamp J (1985) Localization of the Na+/K+ — ATPase and of an amiloride sensitive Na+ uptake on thyroid epithelial cells. Eur J Cell Biol 38:134–141
Gorvel J-P, Rigal A, Sarles J, Maroux S (1985) Aminopeptidase N- and human blood group A-antigenicity along the digestive tract and associated glands in the rabbit. Cell Tissue Res 239:241–248
Herzog V (1983) Transcytosis in the thyroid follicle cell. J Cell Biol 97:607–617
Herzog V, Miller F (1979) Membrane retrieval in epithelial cells of isolated thyroid follicles. Eur J Cell Biol 19:203–215
Hovsepian S, Feracci H, Maroux S, Fayet G (1982) Kinetic studies of the localization of aminopeptidase N in monolayer and in follicle-associated cultures of porcine thyroid cells. Cell Tissue Res 224:601–611
Kenny JA, Maroux S (1982) Topology of microvillar membrane hydrolases of kidney and intestine. Physiol Rev 62:91–127
Kerjaschki D, Noronha-Blob L, Sacktor B, Farquhar MG (1984) Microdomains of distinctive glycoprotein composition in the kidney proximal tubule brush border. J Cell Biol 98:1505–1513
Klausner RD, Kleinfeld AM, Hoover RL, Karnovsky MJ (1980) Lipid domains in membranes: Evidence derived from structural perturbations induced by free fatty acids and lifetime heterogeneity analysis. J Biol Chem 255:1286–1295
Kolb-Bachofen V (1981) Hepatic receptor for asialo-glycoproteins. Ultrastructural demonstration of ligand-induced microaggregation of receptors. Biochim Biophys Acta 645:293–299
Louvard D (1980) Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc Natl Acad Sci USA 77:4132–4136
Louvard D, Maroux S, Vannier Ch, Desnuelle P (1975a) Topological studies on the hydrolases bound to the intestinal brush border membrane. I. Solubilization by papain and Triton X-100. Biochim Biophys Acta 375:236–248
Louvard D, Maroux S, Desnuelle P (1975b) Topological studies on the hydrolases bound to the intestinal brush border membrane. II. Interactions of free and bound aminopeptidase with a specific antibody. Biochim Biophys Acta 389:389–400
Maroux S, Louvard D, Baratti J (1973) The aminopeptidase from hog intestinal brush border. Biochim Biophys Acta 321:282–295
Matsuura S, Eto S, Kato K, Tashiro Y (1984) Ferritin immunoelectron microscopic localization of 5′ nucleotidase on rat liver cell surface. J Cell Biol 99:166–173
Merz WA (1967) Die Streckenmessung an gerichteten Strukturen im Mikroskop und ihre Anwendung zur Bestimmung von Oberflächen-Volumen-Relationen im Knochengewebe. Mikroskopie 22:132–143
Nilsson M, Mölne J, Karlsson AF, Ericson LE (1986) Ultrastructural localization of microsomal antibodies on the surface of human thyroid follicle cells. International Symposium Thyroid and Autoimmunity. Amsterdam, 20–21 March, 1986 [Abstr], no 28
Roman LM, Hubbard AL (1983) A domain-specific marker for the hepatocyte plasma membrane: localization of leucine aminopeptidase to the bile canalicular domain. J Cell Biol 96:1548–1558
Roman LM, Hubbard AL (1984) A domain-specific marker for the hepatocyte plasma membrane. III. Isolation of bile canalicular membrane by immunoadsorption. J Cell Biol 98:1497–1504
Roth J (1982) The protein A-gold (pAu) technique — a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, vol 1. Academic Press, London, pp 107–133
Severs N, Robenek H (1983) Detection of microdomains in biomembranes. An appraisal of recent developments in freeze-fracture cytochemistry. Biochim Biophys Acta 737:373–408
Simionescu N, Simionescu M, Palade GE (1981) Differentiated microdomains on the luminal surface of capillary endothelium. I. Preferential distribution of anionic sites. J Cell Biol 90:605–613
Tice LW, Wollman SH (1974) Ultrastructural localization of peroxidase on pseudopods and other structures of the typical thyroid epithelial cell. Endocrinology 94:1555–1567
Ugolev AM, De Laey P (1973) Membrane digestion. A concept of enzymic hydrolysis on cell membranes. Biochim Biophys Acta 300:105–128
Vannier Ch, Louvard D, Maroux S, Desnuelle P (1976) Structural and topological homology between porcine intestinal and renal brush border aminopeptidase. Biochim Biophys Acta 455:185–199
Westermark K, Westermark B, Karlsson FA, Ericson LE (1986) Location of epidermal growth factor receptors on porcine thyroid follicle cells and receptor regulation by thyrotropin. Endocrinology 118:1040–1046
Wollman SH (1980) Structure of the thyroid gland. In: De Visscher M (ed) The thyroid gland. Raven Press, New York, pp 1–19
Author information
Authors and Affiliations
Additional information
This study was supported by Grant No 12X-537 from the Swedish Medical Research Council
Rights and permissions
About this article
Cite this article
Nilsson, M., Ekholm, R., Fayet, G. et al. Immunocytochemical localization of aminopeptidase N on the cell surface of isolated porcine thyroid follicles. Cell Tissue Res. 250, 295–303 (1987). https://doi.org/10.1007/BF00219074
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00219074