Histochemistry and Cell Biology

, Volume 123, Issue 4–5, pp 529–539 | Cite as

Urothelial injuries and the early wound healing response: tight junctions and urothelial cytodifferentiation

  • Mateja  Erdani Kreft
  • Maksimiljan Sterle
  • Peter Veranič
  • Kristijan Jezernik
Original Paper

Abstract

Using primary explant cultures of mouse bladder, the early response of the urothelium after superficial and full-thickness injuries was investigated. In such an in vitro wound healing model, explant surfaces with a mostly desquamated urothelial superficial layer represented superficial wounds, and the exposed lamina propria at the cut edges of the explants represented full-thickness wounds. The urothelial cell ultrastructure, the expression and subcellular distribution of the tight junctional protein occludin, and differentiation-related proteins CK 20, uroplakins, and actin were followed. Since singular terminally differentiated superficial cells remained on the urothelium after superficial injury (i.e., original superficial cells), we sought to determine their role during the urothelial wound-healing process. Ultrastructural and immunocytochemical studies have revealed that restored tight junctions are the earliest cellular event during the urothelial superficial and full-thickness wound-healing process. Occludin-containing tight junctions are developed before the new superficial cells are terminally differentiated. New insights into the urothelium wound-healing process were provided by demonstrating that the original superficial cells contribute to the urothelium wound healing by developing tight junctions with de novo differentiated superficial cells and by stretching, thus providing a large urothelial surface with asymmetric unit membrane plaques.

Keywords

Urothelial superficial cell Wound healing Tight junctions Differentiation Injury 

References

  1. Bement WM, Forscher P, Mooseker MS (1993) A novel cytoskeletal structure involved in purse string wound closure and cell polarity maintenance. J Cell Biol 121:565–578CrossRefPubMedGoogle Scholar
  2. Fukushima S, Cohen SM., Arai M, Jacobs JB, Friedell GH (1981) Scanning electron microscopic examination of reversible hyperplasia of the rat urinary bladder. Am J Pathol 102:373–380PubMedGoogle Scholar
  3. Hicks RM (1975) The mammalian urinary bladder: an accommodating organ. Biol Rev 50:215–246PubMedGoogle Scholar
  4. Hicks RM, Ketterer B, Warren RC (1974) The ultrastructure and chemistry of the luminal plasma membrane of the mammalian urinary bladder: a structure with low permeability to water and ions. Philos Trans R Soc Lond B 268:23–38Google Scholar
  5. Hu P, Meyers S, Liang F-X, Deng F-M, Kachar B, Zeidel ML, Sun T-T (2002) Role of membrane proteins in permeability barrier function: uroplakin ablation elevates urothelial permeability. Am J Physiol Renal Physiol 283:F1200-F1207PubMedGoogle Scholar
  6. Jezernik K, Romih R, Mannherz HG, Koprivec D (2003) Immunohistochemical detection of apoptosis, proliferation and inducible nitric oxide synthase in rat urothelium damaged by cyclophosphamide treatment. Cell Biol Int 27:863–869CrossRefPubMedGoogle Scholar
  7. Jost SP (1989) Cell cycle of normal bladder urothelium in developing and adult mice. Virchows Arch B Cell Pathol Incl Mol Pathol 57(1):27–36PubMedGoogle Scholar
  8. Kachar B, Liang F, Lins U, Ding M, Wu XR, Stoffler D, Aebi U, Sun T-T (1999) Three-dimensional analysis of the 16 nm urothelial plaque particle: luminal surface exposure, preferential head-to-head interaction, and hinge formation. J Mol Biol 285:595–608CrossRefPubMedGoogle Scholar
  9. Kreft ME, Romih R in Sterle M (2002) Antigenic and ultrastructural markers associated with urothelial cytodifferentiation in primary explant outgrowths of mouse bladder. Int Cell Biol 26:63–74CrossRefGoogle Scholar
  10. Lavelle J, Myers S, Doty D, Buffington A, Zeidel ML, Apodaca G (2000) Urothelial pathophysiological changes in feline interstitial cystitis: a human interstitial cystitis model. Am J Physiol Renal Physiol 278:F540-F553PubMedGoogle Scholar
  11. Lavelle J, Meyers S, Ramage R, Bastacky S, Doty D, Apodaca G, Zeidel ML (2002) Bladder permeability barrier: recovery from selective injury of surface epithelial cells. Am J Physiol Renal Physiol 283:F242-F253PubMedGoogle Scholar
  12. Lewis SA, Diamond JM (1976) Na+ transport by rabbit urinary bladder, a tight epithelium. J Memb Biol 28:1–40Google Scholar
  13. Lotz MM, Rabinovitz I, Mercurio AM (2000) Intestinal restitution: progression of actin cytoskeleton rearrangements and integrin function in a model of epithelial wound healing. Am J Pathol 156:985–995PubMedGoogle Scholar
  14. Matter K, Balda MS (1999) Occludin and the functions of tight junctions. Int Rev Cytol 186:117–146PubMedGoogle Scholar
  15. McCarthy KM, Skare IB, Stankewich MD, Furuse M, Tsukita S (1996) Occludin is a functional component of the tight junction. J Cell Sci 109:2287–2298PubMedGoogle Scholar
  16. Messier B, Leblond CP (1960) Cell proliferation and migration revealed by radioautography after injection of thymidine-H3 into male rats and mice. Am J Anat 106:247–265CrossRefPubMedGoogle Scholar
  17. Min G, Zhou G, Schapira M, Sun T-T, Kong X-P (2003) Structural basis of urothelial permeability barrier function as revealed by Cryo-EM studies of the 16 nm uroplakin particle. J Cell Sci 116:4087–4094CrossRefPubMedGoogle Scholar
  18. Negrete HO, Lavelle JP, Berg J, Lewis SA, and Zeidel ML (1996) Permeability properties of the intact mammalian bladder epithelium. Am J Physiol Renal Fluid Electrolyte Physiol 271:F886-F894Google Scholar
  19. Nusrat A, Delp C, Madara JL (1992) Intestinal epithelial restitution: characterization of cell culture model and mapping of cytoskeletal elements in migrating cells. J Clin Invest 89:1501–1511PubMedGoogle Scholar
  20. Rao JN, Platoshyn O, Li L, Guo X, Golovina VA, Yuan JX-J, Wang J-Y (2002) Activation of K+ channels and increased migration of differentiated intestinal epithelial cells after wounding. Am J Physiol Cell Physiol 282:C885-C898PubMedGoogle Scholar
  21. Romih R, Veranic P, Jezernik K (1999) Actin filaments during terminal differentiation of urothelial cells in the rat urinary bladder. Histochem Cell Biol 112:375–380CrossRefPubMedGoogle Scholar
  22. Romih R, Koprivec D, Martincic DS, Jezernik K (2001) Restoration of the rat urothelium after cyclophosphamide treatment. Cell Biol Int 25:531–537CrossRefPubMedGoogle Scholar
  23. Silen W, Ito S (1985) Mechanism for rapid-epithelialization of the gastric mucosal surface. Annu Rev Physiol 47:217–229CrossRefPubMedGoogle Scholar
  24. Truschel ST, Wang E, Ruiz WG, Leung S-M, Rojas R, Lavelle J, Zeidel M, Stoffer S, Apodaca G (2002) Stretch-regulated exocytosis/endocytosis in bladder umbrella cells. Mol Biol Cell 13(3):830–846CrossRefPubMedGoogle Scholar
  25. Veranič P, Jezernik K (2000) The response of junctional complexes to induced desquamation of mouse bladder urothelium. Biol Cell 92(2):105–113CrossRefPubMedGoogle Scholar
  26. Veranič P, Jezernik K (2002) Trajectorial organisation of cytokeratins within the subapical region of umbrella cells. Cell Motil Cytoskeleton 53(4):317–325CrossRefPubMedGoogle Scholar
  27. Veranič P, Romih R, Jezernik K (2004) What determinates differentiation of urothelial umbrella cells? Eur J Cell Biol 83:27–34PubMedGoogle Scholar
  28. Wu XR, Manabe M, Yu J, Sun T-T (1990) Large scale purification and immunolocalization of bovine uroplakins I, II, and III: molecular markers of urothelial differentiation. J Biol Chem 265:19170–19179PubMedGoogle Scholar
  29. Yu HJ, Chien CT, Lai YJ, Lai MK, Chen CF, Levin RM, Hsu SM (2004) Hypoxia preconditioning attenuates bladder overdistension induced oxidative injury by upregulation of Bcl-2 in the rat. J Physiol 554:815–828CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Mateja  Erdani Kreft
    • 1
  • Maksimiljan Sterle
    • 1
  • Peter Veranič
    • 1
  • Kristijan Jezernik
    • 1
  1. 1.Institute of Cell Biology, Medical FacultyUniversity of LjubljanaLjubljanaSlovenia

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