Summary
The present study describes the intracellular distribution of Tamm-Horsfall protein (THP) in rat kidney. The localization was determined by immunoelectron microscopy using the protein A-gold technique. Various fixation and embedding protocols were evaluated for this purpose. Brief perfusion fixation (3 min) with 1% glutaraldehyde and embedding in a highly hydrophilic glycol methacrylate-polyester mixture were most appropriate for antigen-antibody recognition and structural preservation. The overall tissue distribution of THP was evaluated by indirect immunofluorescence microscopy; reaction was strong along the entire thick ascending limb of the loop of Henle (TAL) with enhanced fluorescence in the apical cytoplasm. On the electron microscopic level immunogold labelling was concentrated over numerous membrane-bound vesicles which form a compartment in the apical cytoplasm. The Golgi region was consistently labelled, whereas the plasma membranes revealed only sporadic labelling at the luminal side, and basolateral membranes were mostly unlabelled. Quantitative evaluation of the gold labelling, which was separately done for the inner stripe, outer stripe and cortical TAL, consistently showed the highest particle density in the apical cytoplasm. Middle and basal levels in the TAL cells were only moderately labelled. The results are discussed with respect to the current opinion which describes THP as a membrane glycoprotein. We speculate that the accumulation of THP in the apical vesicular compartment of TAL cells indicates a storage site of the protein, possibly prior to extrusion via exocytosis of the vesicle contents.
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References
Allen F, Tisher CC (1976) Morphology of the ascending thick limb of Henle. Kidney Int 9:8–22
Greven J, Kölling B, v Bronewski-Schwarzer B, Junker M, Neffgen B, Nilius RM (1984) Evidence for a role of the Tamm-Horsfall protein in the tubular action of furosemide-like loop diuretics. In: Puschett JB (ed) Diuretics. Elsevier Science, Amsterdam, pp 203–214
Hoyer JR (1976) Autoimmune tubulointerstitial nephritis induced in rats by immunization with rat Tamm-Horsfall (TH) urinary glycoprotein (abstr.) Kidney Int 10:544
Hoyer JR (1980) Tubulointerstitial immune complex nephritis in rats immunized with Tamm-Horsfall protein. Kidney Int 17:284–292
Hoyer JR, Seiler MW (1979) Pathophysiology of Tamm-Horsfall protein. Kidney Int 16:279–289
Hoyer JR, Sisson SP, Vernier RL (1978) Immuno-electron microscopic localization of Tamm-Horsfall protein (THP) in rat kidney (abstr). Fed Proc 37:837
Hoyer JR, Sisson SP, Vernier RL (1979) Tamm-Horsfall glycoprotein. Ultrastructural immunoperoxidase localization in rat kidney. Lab Invest 41:168–173
Kaissling B, Kriz W (1979) Structural analysis of the rabbit kidney. Adv Anat Embryol Cell Biol 56:1–123
Kaissling B, Peter S, Kriz W (1977) Transition of the thick ascending limb of Henle's loop into the distal convoluted tubule in the nephron of the rat kidney. Cell Tissue Res 182:111–118
Karnovsky MJ (1971) Use of ferrocyanide-reduced osmium tetroxide in electron microscopy (abstr). J Cell Biol 51:146a
Koeppen-Hagemann I, Bachmann S, Minuth WW, Kriz W (1984) Immunohistochemical localization of a protein fraction derived from rabbit renal papilla. Histochemistry 81:457–464
Kone BC, Madsen K, Tisher CC (1984) Ultrastructure of the thick ascending limb of Henle in the rat kidney. Am J Anat 171:217–226
McLean JW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083
Roth J, Bendayan M, Orci L (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex. J Histochem Cytochem 26:1074–1081
Roth J, Bendayan M, Carlemalm E, Villiger W, Garavito M (1981) Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J Histochem Cytochem 29:663–671
Seiler MW, Hoyer JR (1981) Ultrastructural studies of tubulointerstitial immune complex nephritis in rats immunized with Tamm Horsfall protein. Lab Invest 45:321–327
Sikri KL, Foster CL, Bloomfield FJ, Marshall RD (1979) Localization by immunofluorescence and by light- and electron microscopic immunoperoxidase techniques of Tamm-Horsfall glycoprotein in adult hamster kidney. Biochem J 181:525–532
Sikri KL, Foster CL, MacHugh N, Marshall RD (1981) Localization of Tamm-Horsfall glycoprotein in the human kidney using immunofluorescence and immuno-electron microscopical techniques. J Anat 132:597–605
Sikri KL, Alexander DP, Foster CL (1982) Localization of Tamm-Horsfall glycoprotein in the normal rat kidney and the effect of adrenalectomy on its localization in the hamster and rat kidney. J Anat 135:29–45
Sisson S, Vernier RL (1980) Methods for immunoelectron microscopy: localization of antigens in rat kidney. J Histochem Cytochem 28:441–452
Stevenson FK, Cleave AJ, Kent PW (1970) The effect of ions on the viscometric and ultracentrifugal behaviour of Tamm-Horsfall glycoprotein. Biochim Biophys Acta 236:59–66
Tamm I, Horsfall FL (1952) A mucoprotein derived from human urine which reacts with influenza, mumps, and Newcastle disease viruses. J Exp Med 95:71–97
Tisher CC (1981) Anatomy of the kidney. In: Brenner MB, Rector FC (eds) The Kidney, Vol 1. WB Saunders, Philadelphia, pp 3–75
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
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Bachmann, S., Koeppen-Hagemann, I. & Kriz, W. Ultrastructural localization of Tamm-Horsfall glycoprotein (THP) in rat kidney as revealed by protein A-gold immunocytochemistry. Histochemistry 83, 531–538 (1985). https://doi.org/10.1007/BF00492456
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DOI: https://doi.org/10.1007/BF00492456