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. 1995 Dec;96(6):2646–2653. doi: 10.1172/JCI118330

Clusterin promotes the aggregation and adhesion of renal porcine epithelial cells.

J R Silkensen 1, K M Skubitz 1, A P Skubitz 1, D H Chmielewski 1, J C Manivel 1, J A Dvergsten 1, M E Rosenberg 1
PMCID: PMC185970  PMID: 8675630

Abstract

The function of clusterin, a heterodimeric glycoprotein markedly induced in renal and other organ injuries, is unclear. Since renal injury is accompanied by alterations in cell attachment, it is possible that clusterin functions to promote cell-cell and cell-substratum interactions. In this study, a single cell suspension of renal epithelial (LLC-PK1) cells was treated with purified human clusterin, resulting in time- and dose-dependent cell aggregation. Electron microscopy of the cell aggregates demonstrated cell junction and lumen formation. To determine the effect of clusterin on cell adhesion, tissue culture plates were coated with clusterin, fibronectin, PBS, or albumin. Clusterin and fibronectin promoted cell adhesion to the same extent. The adhesion to clusterin was dose dependent and specific, as a monoclonal antibody against clusterin inhibited cell adhesion to clusterin but not fibronectin. Perterbations of the cytoskeleton may underlie the alterations in cell attachment which occur in renal injury. Induction of clusterin mRNA was seen after disruption of both microtubules and microfilaments and after inhibition of cell-substratum interactions. In conclusion, clusterin is a potent renal epithelial cell aggregation and adhesion molecule. We speculate that clusterin functions to promote cell-cell and cell-substratum interactions which are perturbed in the setting of renal injury, thereby preserving the integrity of the renal epithelial barrier.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aronow B. J., Lund S. D., Brown T. L., Harmony J. A., Witte D. P. Apolipoprotein J expression at fluid-tissue interfaces: potential role in barrier cytoprotection. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):725–729. doi: 10.1073/pnas.90.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bacallao R., Fine L. G. Molecular events in the organization of renal tubular epithelium: from nephrogenesis to regeneration. Am J Physiol. 1989 Dec;257(6 Pt 2):F913–F924. doi: 10.1152/ajprenal.1989.257.6.F913. [DOI] [PubMed] [Google Scholar]
  3. Blaschuk O., Burdzy K., Fritz I. B. Purification and characterization of a cell-aggregating factor (clusterin), the major glycoprotein in ram rete testis fluid. J Biol Chem. 1983 Jun 25;258(12):7714–7720. [PubMed] [Google Scholar]
  4. Bonventre J. V. Mechanisms of ischemic acute renal failure. Kidney Int. 1993 May;43(5):1160–1178. doi: 10.1038/ki.1993.163. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Calvet J. P. Polycystic kidney disease: primary extracellular matrix abnormality or defective cellular differentiation? Kidney Int. 1993 Jan;43(1):101–108. doi: 10.1038/ki.1993.17. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Correa-Rotter R., Hostetter T. H., Manivel J. C., Eddy A. A., Rosenberg M. E. Intrarenal distribution of clusterin following reduction of renal mass. Kidney Int. 1992 Apr;41(4):938–950. doi: 10.1038/ki.1992.144. [DOI] [PubMed] [Google Scholar]
  9. Correa-Rotter R., Mariash C. N., Rosenberg M. E. Loading and transfer control for Northern hybridization. Biotechniques. 1992 Feb;12(2):154–158. [PubMed] [Google Scholar]
  10. Diemer V., Hoyle M., Baglioni C., Millis A. J. Expression of porcine complement cytolysis inhibitor mRNA in cultured aortic smooth muscle cells. Changes during differentiation in vitro. J Biol Chem. 1992 Mar 15;267(8):5257–5264. [PubMed] [Google Scholar]
  11. Dvergsten J., Manivel J. C., Correa-Rotter R., Rosenberg M. E. Expression of clusterin in human renal diseases. Kidney Int. 1994 Mar;45(3):828–835. doi: 10.1038/ki.1994.109. [DOI] [PubMed] [Google Scholar]
  12. French L. E., Chonn A., Ducrest D., Baumann B., Belin D., Wohlwend A., Kiss J. Z., Sappino A. P., Tschopp J., Schifferli J. A. Murine clusterin: molecular cloning and mRNA localization of a gene associated with epithelial differentiation processes during embryogenesis. J Cell Biol. 1993 Sep;122(5):1119–1130. doi: 10.1083/jcb.122.5.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. French L. E., Tschopp J., Schifferli J. A. Clusterin in renal tissue: preferential localization with the terminal complement complex and immunoglobulin deposits in glomeruli. Clin Exp Immunol. 1992 Jun;88(3):389–393. doi: 10.1111/j.1365-2249.1992.tb06459.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fritz I. B., Burdzy K. Novel action of carnitine: inhibition of aggregation of dispersed cells elicited by clusterin in vitro. J Cell Physiol. 1989 Jul;140(1):18–28. doi: 10.1002/jcp.1041400104. [DOI] [PubMed] [Google Scholar]
  15. Fritz I. B., Burdzy K., Sétchell B., Blaschuk O. Ram rete testis fluid contains a protein (clusterin) which influences cell-cell interactions in vitro. Biol Reprod. 1983 Jun;28(5):1173–1188. doi: 10.1095/biolreprod28.5.1173. [DOI] [PubMed] [Google Scholar]
  16. Han J. R., Suiko M., Liu C. C., Liu M. C. Post-translational modifications and binding properties of the apically secreted 80-kDa glycoprotein from Madin-Darby canine kidney cells: similarities to the C-terminal portion of the basolaterally secreted fibronectin. Arch Biochem Biophys. 1991 May 1;286(2):337–345. doi: 10.1016/0003-9861(91)90049-o. [DOI] [PubMed] [Google Scholar]
  17. Harding M. A., Chadwick L. J., Gattone V. H., 2nd, Calvet J. P. The SGP-2 gene is developmentally regulated in the mouse kidney and abnormally expressed in collecting duct cysts in polycystic kidney disease. Dev Biol. 1991 Aug;146(2):483–490. doi: 10.1016/0012-1606(91)90249-3. [DOI] [PubMed] [Google Scholar]
  18. Jenne D. E., Tschopp J. Clusterin: the intriguing guises of a widely expressed glycoprotein. Trends Biochem Sci. 1992 Apr;17(4):154–159. doi: 10.1016/0968-0004(92)90325-4. [DOI] [PubMed] [Google Scholar]
  19. Jenne D. E., Tschopp J. Molecular structure and functional characterization of a human complement cytolysis inhibitor found in blood and seminal plasma: identity to sulfated glycoprotein 2, a constituent of rat testis fluid. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7123–7127. doi: 10.1073/pnas.86.18.7123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jordan-Starck T. C., Lund S. D., Witte D. P., Aronow B. J., Ley C. A., Stuart W. D., Swertfeger D. K., Clayton L. R., Sells S. F., Paigen B. Mouse apolipoprotein J: characterization of a gene implicated in atherosclerosis. J Lipid Res. 1994 Feb;35(2):194–210. [PubMed] [Google Scholar]
  21. Kirszbaum L., Bozas S. E., Walker I. D. SP-40,40, a protein involved in the control of the complement pathway, possesses a unique array of disulphide bridges. FEBS Lett. 1992 Feb 3;297(1-2):70–76. doi: 10.1016/0014-5793(92)80330-j. [DOI] [PubMed] [Google Scholar]
  22. Kirszbaum L., Sharpe J. A., Murphy B., d'Apice A. J., Classon B., Hudson P., Walker I. D. Molecular cloning and characterization of the novel, human complement-associated protein, SP-40,40: a link between the complement and reproductive systems. EMBO J. 1989 Mar;8(3):711–718. doi: 10.1002/j.1460-2075.1989.tb03430.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kounnas M. Z., Loukinova E. B., Stefansson S., Harmony J. A., Brewer B. H., Strickland D. K., Argraves W. S. Identification of glycoprotein 330 as an endocytic receptor for apolipoprotein J/clusterin. J Biol Chem. 1995 Jun 2;270(22):13070–13075. doi: 10.1074/jbc.270.22.13070. [DOI] [PubMed] [Google Scholar]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. May P. C., Finch C. E. Sulfated glycoprotein 2: new relationships of this multifunctional protein to neurodegeneration. Trends Neurosci. 1992 Oct;15(10):391–396. doi: 10.1016/0166-2236(92)90190-j. [DOI] [PubMed] [Google Scholar]
  26. Michel D., Chabot J. G., Moyse E., Danik M., Quirion R. Possible functions of a new genetic marker in central nervous system: the sulfated glycoprotein-2 (SGP-2). Synapse. 1992 Jun;11(2):105–111. doi: 10.1002/syn.890110203. [DOI] [PubMed] [Google Scholar]
  27. Millis A. J., Hoyle M., Kent L. In vitro expression of a 38,000 dalton heparin-binding glycoprotein by morphologically differentiated smooth muscle cells. J Cell Physiol. 1986 Jun;127(3):366–372. doi: 10.1002/jcp.1041270304. [DOI] [PubMed] [Google Scholar]
  28. Molitoris B. A., Nelson W. J. Alterations in the establishment and maintenance of epithelial cell polarity as a basis for disease processes. J Clin Invest. 1990 Jan;85(1):3–9. doi: 10.1172/JCI114427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Molitoris B. A. New insights into the cell biology of ischemic acute renal failure. J Am Soc Nephrol. 1991 Jun;1(12):1263–1270. doi: 10.1681/ASN.V1121263. [DOI] [PubMed] [Google Scholar]
  30. Murphy B. F., Davies D. J., Morrow W., d'Apice A. J. Localization of terminal complement components S-protein and SP-40,40 in renal biopsies. Pathology. 1989 Oct;21(4):275–278. doi: 10.3109/00313028909061073. [DOI] [PubMed] [Google Scholar]
  31. Murphy B. F., Kirszbaum L., Walker I. D., d'Apice A. J. SP-40,40, a newly identified normal human serum protein found in the SC5b-9 complex of complement and in the immune deposits in glomerulonephritis. J Clin Invest. 1988 Jun;81(6):1858–1864. doi: 10.1172/JCI113531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Purrello M., Bettuzzi S., Di Pietro C., Mirabile E., Di Blasi M., Rimini R., Grzeschik K. H., Ingletti C., Corti A., Sichel G. The gene for SP-40,40, human homolog of rat sulfated glycoprotein 2, rat clusterin, and rat testosterone-repressed prostate message 2, maps to chromosome 8. Genomics. 1991 May;10(1):151–156. doi: 10.1016/0888-7543(91)90495-z. [DOI] [PubMed] [Google Scholar]
  33. Racusen L. C., Fivush B. A., Li Y. L., Slatnik I., Solez K. Dissociation of tubular cell detachment and tubular cell death in clinical and experimental "acute tubular necrosis". Lab Invest. 1991 Apr;64(4):546–556. [PubMed] [Google Scholar]
  34. Rocco M. V., Neilson E. G., Hoyer J. R., Ziyadeh F. N. Attenuated expression of epithelial cell adhesion molecules in murine polycystic kidney disease. Am J Physiol. 1992 Apr;262(4 Pt 2):F679–F686. doi: 10.1152/ajprenal.1992.262.4.F679. [DOI] [PubMed] [Google Scholar]
  35. Rosenberg M. E., Dvergsten J., Correa-Rotter R. Clusterin: an enigmatic protein recruited by diverse stimuli. J Lab Clin Med. 1993 Feb;121(2):205–214. [PubMed] [Google Scholar]
  36. Rosenberg M. E., Manivel J. C., Carone F. A., Kanwar Y. S. Genesis of renal cysts is associated with clusterin expression in experimental cystic disease. J Am Soc Nephrol. 1995 Mar;5(9):1669–1674. doi: 10.1681/ASN.V591669. [DOI] [PubMed] [Google Scholar]
  37. Rosenberg M. E., Silkensen J. Clusterin and the kidney. Exp Nephrol. 1995 Jan-Feb;3(1):9–14. [PubMed] [Google Scholar]
  38. Thomas-Salgar S., Millis A. J. Clusterin expression in differentiating smooth muscle cells. J Biol Chem. 1994 Jul 8;269(27):17879–17885. [PubMed] [Google Scholar]
  39. Tobe T., Minoshima S., Yamase S., Choi N. H., Tomita M., Shimizu N. Assignment of a human serum glycoprotein SP-40,40 gene (CLI) to chromosome 8. Cytogenet Cell Genet. 1991;57(4):193–195. doi: 10.1159/000133144. [DOI] [PubMed] [Google Scholar]
  40. Tsuruta J. K., Wong K., Fritz I. B., Griswold M. D. Structural analysis of sulphated glycoprotein 2 from amino acid sequence. Relationship to clusterin and serum protein 40,40. Biochem J. 1990 Jun 15;268(3):571–578. doi: 10.1042/bj2680571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tung P. S., Burdzy K., Wong K., Fritz I. B. Competition between cell-substratum interactions and cell-cell interactions. J Cell Physiol. 1992 Aug;152(2):410–421. doi: 10.1002/jcp.1041520224. [DOI] [PubMed] [Google Scholar]
  42. Väkevä A., Laurila P., Meri S. Co-deposition of clusterin with the complement membrane attack complex in myocardial infarction. Immunology. 1993 Oct;80(2):177–182. [PMC free article] [PubMed] [Google Scholar]
  43. Weinberg J. M. The cell biology of ischemic renal injury. Kidney Int. 1991 Mar;39(3):476–500. doi: 10.1038/ki.1991.58. [DOI] [PubMed] [Google Scholar]
  44. Witte D. P., Aronow B. J., Stauderman M. L., Stuart W. D., Clay M. A., Gruppo R. A., Jenkins S. H., Harmony J. A. Platelet activation releases megakaryocyte-synthesized apolipoprotein J, a highly abundant protein in atheromatous lesions. Am J Pathol. 1993 Sep;143(3):763–773. [PMC free article] [PubMed] [Google Scholar]
  45. Wong P., Pineault J., Lakins J., Taillefer D., Léger J., Wang C., Tenniswood M. Genomic organization and expression of the rat TRPM-2 (clusterin) gene, a gene implicated in apoptosis. J Biol Chem. 1993 Mar 5;268(7):5021–5031. [PubMed] [Google Scholar]
  46. You Y., Hirsch D. J., Morgunov N. S. Functional integrity of proximal tubule cells. Effects of hypoxia and ischemia. J Am Soc Nephrol. 1992 Oct;3(4):965–974. doi: 10.1681/ASN.V34965. [DOI] [PubMed] [Google Scholar]

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