Summary
Localization of cathepsin L in rat kidney was investigated by immunocytochemical techniques. Kidneys were fixed by perfusion and embedded in Epon or Lowicryl K4M without postomication. For light microscopy (LM), semi-thin sections of the Epon-embedded material were stained by the immunoenzyme technique after removal of epoxy resin. For electron microscopy (EM), ultra-thin sections of Lowicryl K4M-embedded material were stained by the protein A-gold technique. By LM, reaction deposits for cathepsin L were present in the cytoplasmic granules of proximal tubule cells, but little or no reaction product was noted in distal tubule, collecting tubule, and most of urinary tubules in the medulla. By EM, heavy gold label for cathepsin L was confined exclusively to lysosomes of the proximal tubule cells, but little or no label to those of the other segments. In immunocytochemical control sections, no reaction was observed. These results indicate that a main container of cathepsin L is lysosomes of the proximal tubule and suggest that the enzyme plays a role in the degradation of endocytosed proteins.
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References
Bando Y, Kominami E, Katunuma N (1986) Purification and tissue distribution of rat cathepsin L. J Biochem 100:35–42
Ericsson JLE (1965) Transport and digestion of hemoglobin in the proximal tubule. II. Electron microscopy. Lab Invest 14:16–39
Erikson AH, Blobel G (1979) Early events in the biosynthesis of the lysosomal enzyme cathepsin D. J Biol Chem 254:11771–11774
Ii K, Hizawa K, Kominami E, Bando Y, Katunuma N (1985) Different immunolocalizations of cathepsins B, H and L in the liver. J Histochem Cytochem 33:1173–1175
Kirschke H, Langner J, Wiederander B, Ansorge S, Bohley P (1977) Cathepsin L. A new proteinase from rat-liver lysosomes. Eur J Biochem 74:293–301
Kirschke H, Langner J, Riemann S, Wiederanders B, Ansorge S, Bohley P (1980) Lysosomal cystein proteinases. In: Evered D, Whelan J (eds) Protein degradation of in health and disease. Excerpta Medica, Amsterdam, pp 15–35
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–684
Litwin JA, Yokota S, Hashimoto T, Fahimi HD (1984) Light microscopic immunocytochemical demonstration of peroxisomal enzymes in Epon sections. Histochemistry 81:15–22
Mason RW, Taylor MAJ, Etherington DJ (1984) The purification and properties of cathepsin L from rabbit liver. Biochem J 217:209–217
Mason RW, Green GDJ, Barrett AJ (1985) Human liver cathepsin L. Biochem J 226:233–241
Marx R, Mölbert E, Zollinger HU (1966) Elektronenmikroskopische Untersuchungen am proximalen Tubules der Mäuseniere bei Eiweißspeicherung. Virchows Arch Pathol Anat Physiol 341:9–23
Maunsbach AB (1966) Albumin absorption by renal proximal tubule cells. Nature 212:546–547
Maunsbach AB (1973) Ultrastructure of the proximal tubule. In: Orloff J, Berliner RW (eds) Handbook of physiology, sect 8, renal physiology. American Physiological Society, Washington, pp 31–79
Miller F, Palade GE (1964) Lytic activities in renal protein absorption droplets. An electron microscopical cytochemical study. J Cell Biol 23:519–552
Rosenfeld MG, Kreibich G, Popov K, Kato K, Sabatini DD (1982) Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and role of co- and post-translational processing in determining their subcellular distribution. J Cell Biol 93:135–143
Roth J, Bendayan M, Carlemalm E, Villiger W Garavito M (1981) The enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J Histochem Cytochem 29:663–671
Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol 90:533–536
Sly WS, Fischer HD, Gonzalez-Noriega A, Grubb JH, Natowicz M (1981) Role of the 6-phosphomannosyl-enzyme receptor in intracellular transport and absorptive pinocytosis of lysosomal enzymes. In: Hand AR, Oliver C (eds) Methods in cell biology, vol 23. Academic Press, New York, pp 191–214
Stathis EC, Fabrikanos A (1958) Preparation of colloidal gold. Chem Ind (London) 27:860–861
Straus W (1962) Colorimetric investigation of uptake of an intravenously injected protein (horseradish peroxidase) by rat kidney and effects of competition by egg white. J Cell Biol 12:231–246
Straus W (1964) Occurrence of phagosomes and phago-lysosomes in different segments of the nephron in relation to reabsorption, transport, digestion and extrusion of intravenously injected horseradish peroxidase. J Cell Biol 21:295–308
Tanaka K, Ikegaki N, Ichihara A (1984) Purification and characterization of hemoglobin-hydrolyzing acidic thiol proteinase induced by leupeptin in rat liver. J Biol Chem 259:5937–5944
Thoenes W, Langer KH (1969) Die Endocytose-Phase der Eiweiß-resorption im proximalen Nierentubulus. Virchows Arch Pathol Anat Physiol 2:361–379
Tisher CC (1981) Anatomy of the kidney. In: Brenner BM, Rector FC (eds) The kidney, vol 1. WB Saunders, Philadelphia, pp 3–75
Towatari T, Tanaka K, Yoshikawa D, Katunuma N (1978) Purification and properties of a new cathepsin from rat liver. J Biochem 84:659–671
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
van Dongen JM, Barnevelt RA, Geuze HJ, (1984) Immunocytochemistry of lysosomal hydrolases and their precursor forms in normal and mutant human cells. Histochem J 16:941–954
Yokota S, Kato K (1988a) Involvement of cathepsins B and H in lysosomal degradation of horseradish peroxidase endocytosed by the proximal tubule cells of rat kidney. I. Histochemical and immunohistochemical studies. Anat Rec (in press)
Yokota S, Kato K (1988b) Involvement of cathepsins B and H in lysosomal degradation of horseradish peroxidase endocytosed by the proximal tubule cells of rat kidney. II. Immunocytochemical studies using protein A-gold technique applied to conventional and serial sections. Anat Rec (in press)
Yokota S, Tsuji H, Kato K (1985) Immunocytochemical localization of cathepsin D in lysosomes of cortical collecting tubule cells of the rat kidney. J Histochem Cytochem 33:191–200
Yokota S, Tsuji H, Kato K (1986a) Immunocytochemical localization of cathepsin H in rat kidney. Light and electron microscopic study. Histochemistry 85:223–230
Yokota S, Tsuji H, Kato K (1986b) Immunocytochemical localization of cathepsin B in rat kidney. I. Light microscopic study using the indirect immunoenzyme technique. J Histochem Cytochem 34:891–897
Yokota S, Tsuji H, Kato K (1986c) Immunocytochemical localization of cathepsin B in rat kidney. II. Electron microscopic study using the protein A-gold technique. J Histochem Cytochem 34:899–907
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Yokota, S., Nishimura, Y. & Kato, K. Localization of cathepsin L in rat kidney revealed by immunoenzyme and immunogold techniques. Histochemistry 90, 277–283 (1988). https://doi.org/10.1007/BF00495971
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DOI: https://doi.org/10.1007/BF00495971