Advertisement

Histochemistry

, Volume 82, Issue 2, pp 141–148 | Cite as

Localization of cathepsin D in rat liver

Immunocytochemical study using post-embedding immunoenzyme and protein A-gold techniques
  • S. Yokota
  • H. Tsuji
  • K. Kato
Article

Summary

Light and electron microscopic localization of cathepsin D in rat liver was investigated by post-embedding immunoenzyme and protein A-gold techniques. By light microscopy, cytoplasmic granules of parenchymal cells and Kupffer cells were stained for cathepsin D. Weak staining was also noted in sinusoidal endothelial cells. In the parenchymal cells many of positive granules located around bile canaliculi. In the Kupffer cells and the endothelial cells, diffuse staining was noted in the cytoplasm in addition to granular staining. By electron microscopy, gold particles representing the antigenic sites for cathepsin D were seen in typical secondary lysosomes and some multivesicular bodies of the parenchymal cells and Kupffer cells. The lysosomes of the endothelial cells and fat-storing cells were weakly labeled. Quantitative analysis of the labeling density in the lysosomes of these three types of cells demonstrated that the lysosomes of parenchymal cells and Kupffer cells are main containers of cathepsin D in rat liver. The results suggest that cathepsin D functions in the intracellular digestive system of parenchymal cells and Kupffer cells but not so much in that of the endothelial cells.

Keywords

Endothelial Cell Gold Particle Parenchymal Cell Kupffer Cell Antigenic Site 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Avrameas S, Ternynck T (1971) Peroxidase labeled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry 8:1175–1179Google Scholar
  2. Barrett AJ (1970) Cathepsin D purification of isoenzymes from human and chicken liver. Biochem J 117:601–607Google Scholar
  3. Barrett AJ (1977) Cathepsin D and other carboxyl proteinase. In: Barrett AJ (ed) Proteinases and mammalian cells and tissues. North-Holland, Amsterdam, pp 209–248Google Scholar
  4. Barrett AJ (1978) Lysosomal proteinases and their specificity. In: Segal HL, Doyle DJ (eds) Protein turnover and lysosome function. Academic Press, New York, pp 295–304Google Scholar
  5. Benuck M, Grynbaum A, Marks N (1977) Breakdown of somatostatin and substance P by cathepsin D purified from calf brain by affinity chromatography. Brain Res 143:181–185Google Scholar
  6. Bird JWC, Spanier AM, Schwartz WN (1978) Cathepsin B and D: Proteolytic activity and ultrastructural localization in skeletal muscle. In: Segal HL, Doyle DJ (eds) Protein turnover and lysosome function. Academic Press, New York, pp 589–604Google Scholar
  7. Buys CHCM, Elferink MGL, Bouma JMW, Gruber M (1973) Proteolysis of formaldehyde-treated albumin in Kupffer cells and its inhibition by suramin. J Reticuloendothel Soc 14:209–223Google Scholar
  8. Buys CHCM, De Jong ASH, Bouma JMW, Gruber M (1975) Rapid uptake by liver sinusoidal cells of serum albumin modified with retention of its compact conformation. Biochim Biophys Acta 392:95–100Google Scholar
  9. Erickson AH, Blobel G (1979) Early events in the biosynthesis of the lysosomal enzyme cathepsin D. J Biol Chem 254:11771–11774Google Scholar
  10. Fahimi HD (1967) Perfusion and immersion fixation of rat liver with glutaraldehyde. Lab Invest 16:736–750Google Scholar
  11. Graham RC, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302Google Scholar
  12. Kishida Y, Olsen BR, Berg RA, Prockop DJ (1975) Two improved methods for preparing ferritin-protein conjugates for electron microscopy. J Cell Biol 64:331–339Google Scholar
  13. Knook DL (1977) The role of lysosomal enzymes in protein degradation in different types of rat liver cells. Acta Biol Med Germ 36:1747–1752Google Scholar
  14. Kooistra T, Duursma A, Bouma JMW, Gruber M (1977) Endocytosis and breakdown of proteins by sinusoidal liver cells. Acta Biol Med Germ 36:1763–1776Google Scholar
  15. Lazarus GS, Poole AR (1975) Immunocytochemical localization of Cathepsin D in rabbit skin. Arch Dermatol 111:1150–1153Google Scholar
  16. Litwin JA, Yokota S, Fahimi HD (1984) Light microscopic immunocytochemical demonstration of peroxisomal enzymes in Epon sections. Histochemistry 81:15–22Google Scholar
  17. Mort TS, Poole AR, Decker RS (1981) Immunofluorescent localization of cathepsin B and D in human fibroblasts. J Histochem Cytochem 29:649–657Google Scholar
  18. Nilsson M, Berg T (1977) Uptake and degradation of formaldehyde-treated125I-labelled human serum albumin in rat liver cells in vitro and in vivo. Biochim Biophys Acta 497:171–182Google Scholar
  19. Ogunro EA, Lanman RB, Spencer JR, Ferguson AG, Lesch M (1979) Degradation of canine myosin and actin by cathepsin D isolated from homologous tissue. Cardiovasc Res 13:621–629Google Scholar
  20. Poole AR (1977) Antibodies to enzymes and their uses with particular reference to lysosomal enzymes. In: Dingle JT (ed) Lysosomes: a laboratory handbook. Elsevier/North-Holland Biochemical Press, Amsterdam, pp 245–312Google Scholar
  21. Poole AR, Dingle JT, Barrett AJ (1972) The immunocytochemical demonstration of cathepsin D. J Histochem Cytochem 20:261–265Google Scholar
  22. Rojas-Espinosa O, Dannenberg AM, Sternberg LA, Tsuda T (1974) The role of cathepsin D in the pathogenesis of tuberculosis. A histochemical study employing unlabeled antibodies and the peroxidase-antiperoxidase complex. Am J Pathol 74:1–17Google Scholar
  23. Rosenfeld MG, Kreibich G, Popov K, Kato K, Sabatini DD (1982) Biosynthesis of lysosomal hydrolases: Their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution. J Cell Biol 93:135–143Google Scholar
  24. 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 29:663–671Google Scholar
  25. Schwartz WN, Bird JWC (1977) Degradation of myofibrilar proteins by cathepsin B and D. Biochem J 167:811–820Google Scholar
  26. Slot JW, Geuze HJ (1981) Sizing of protein A-gold probe for immunoelectron microscopy. J Cell Biol 90:533–536Google Scholar
  27. Sly WS, Fisher HD, Gonzalez-Noriega A, Grubb JH, Natowicz M (1981) Role of the 6-phosphomannosyl-enzyme receptor in intracellular transport and adsorptive pinocytosis of lysosomal enzymes. In: Hand AR, Oliver C (eds) Methods in cell biology, vol 23. Academic Press, New York, pp 191–214Google Scholar
  28. Stathis EC, Fabrikanos A (1958) Preparation of colloidal gold. Chem Ind (London) 27:860–861Google Scholar
  29. Van Berkel, TJC (1979) The role of non-parenchymal cells in liver metabolism. Trends Biochem Sci 4:202–205Google Scholar
  30. Van Berkel TJC, Kruijt JK, Koster JF (1975) Identity and activities of lysosomal enzymes in parenchymal and non-parenchymal cells from rat liver. Eur J Biochem 58:145–152Google Scholar
  31. Weibel ER, Kistler GS, Scherle WF (1966) Practical stereological methods for light and electron microscopy. J Cell Biol 30:23–38Google Scholar
  32. Whitaker JN, Seyer JM (1979) Sequential limited degradation of myelin basic protein by cathepsin D. J Biol Chem 254:6956–6963Google Scholar
  33. Whitaker PB, Rhodes RH (1983) The distribution of cathepsin D in rat tissues determined by immunocytochemistry. Am J Anat 166:417–428Google Scholar
  34. Whitaker JN, Terry LC, Whetsell Jr WO (1981) Immunocytochemical localization of cathepsin D in rat neural tissue. Brain Res 216:109–124Google Scholar
  35. Yamamoto K, Kamata O, Katsuda N, Kato K (1980) Immunochemical difference between cathepsin D and cathepsin E-like enzyme from rat spleen. J Biochem (Tokyo) 87:511–516Google Scholar
  36. Yokota S, Atsumi S (1983) Immunoelectron microscopic localization of cathepsin D in lysosomes of rat nerve cells. Histochemistry 79:345–352Google Scholar
  37. Yokota S, Fahimi HD (1984) Receptor-mediated endocytosis of denaturated homologous albumin by sinus lining cells of rat liver. In: Seno S, Okada Y (eds) International cell biology 1984, The Japan Society for Cell Biology, pp 332 (Abstract)Google Scholar
  38. Yokota S, Tsuji H, Kato K (1984) Localization of lysosomal and peroxisomal enzymes in the specific granules of rat intestinal eosinophil leukocytes revealed by immunoelectron microscopic techniques. J Histochem Cytochem 32:267–274Google Scholar
  39. Yokota S, Tsuiji H, Kato K (1984) Immunocytochemical localization of cathepsin D in lysosomes of cortical collecting tubule cells of the rat kidney. J Histochem Cytochem (in press)Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • S. Yokota
    • 1
  • H. Tsuji
    • 2
  • K. Kato
    • 2
  1. 1.Department of AnatomyYamanashi Medical SchoolYamanashiJapan
  2. 2.Department of Physiological Chemistry, Faculty of Pharmaceutical ScienceKyushu UniversityFukuokaJapan

Personalised recommendations