Histochemistry and Cell Biology

, Volume 123, Issue 4–5, pp 335–346 | Cite as

Characterization of renal interstitial fibroblast-specific protein 1/S100A4-positive cells in healthy and inflamed rodent kidneys

  • Michel Le Hir
  • Ivan Hegyi
  • Dominique Cueni-Loffing
  • Johannes Loffing
  • Brigitte Kaissling
Original Paper

Abstract

Fibrosis is considered as a central factor in the loss of renal function in chronic kidney diseases. The origin of fibroblasts and myofibroblasts that accumulate in the interstitium of the diseased kidney is still a matter of debate. It has been shown that accumulation of myofibroblasts in inflamed and fibrotic kidneys is associated with upregulation of fibroblast-specific protein 1 (FSP1, S100A4), not only in the renal interstitium but also in the injured renal epithelia. The tubular expression of FSP1 has been taken as evidence of myofibroblast formation by epithelial–mesenchymal transition (EMT). The identity of FSP1/S100A4 cells has not been defined in detail. We originally intended to use FSP1/S100A4 as a marker of putative EMT in a model of distal tubular injury. However, since the immunoreactivity of FSP1 did not seem to fit with the distribution and shape of fibroblasts or myofibroblasts, we undertook the characterization of FSP1/S100A4-expressing cells in the interstitium of rodent kidneys. We performed immunolabeling for FSP1/S100A4 on thin cryostat sections of perfusion-fixed rat and mouse kidneys with peritubular inflammation, induced by thiazides and glomerulonephritis, respectively, in combination with ecto-5′-nucleotidase (5′NT), recognizing local cortical peritubular fibroblasts, with CD45, MHC class II, CD3, CD4 and Thy 1, recognizing mononuclear cells, with alpha smooth muscle actin (αSMA), as marker for myofibroblasts, and vimentin for intracellular intermediate filaments in cells of mesenchymal origin. In the healthy interstitium of rodents the rare FSP1/S100A4+ cells consistently co-expressed CD45 or lymphocyte surface molecules. Around the injured distal tubules of rats treated for 3–4 days with thiazides, FSP1+/S100A4+, 5′NT+, αSMA+, CD45+ and MHC class II+ cells accumulated. FSP1+/S100A4+ cells consistently co-expressed CD45. In the inflamed regions, αSMA was co-expressed by 5′NT+ cells. In glomerulonephritic mice, FSP1+/S100A4+ cells co-expressed Thy 1, CD4 or CD3. Thus, in the inflamed interstitium around distal tubules of rats and of glomerulonephritic mice, the majority of FSP1+ cells express markers of mononuclear cells. Consequently, the usefulness of FSP1/S100A4 as a tool for detection of (myo)fibroblasts in inflamed kidneys and of EMT in vivo is put into question. In the given rat model the consistent co-expression of αSMA and 5′NT suggests that myofibroblasts originate from resident peritubular fibroblasts.

Keywords

Kidney Fibroblast Myofibroblast Epithelial mesenchymal transition Thiazide FSP1 S100A4 Renal interstitium 

References

  1. Abbate M, Zoja C, Rottoli D, Corna D, Tomasoni S, Remuzzi G (2002) Proximal tubular cells promote fibrogenesis by TGF-beta1-mediated induction of peritubular myofibroblasts. Kidney Int 61:2066–2077Google Scholar
  2. Alpers CE, Hudkins KL, Floege J, Johnson RJ (1994) Human renal cortical interstitial cells with some features of smooth muscle cells participate in tubulointerstitial and crescentic glomerular injury. J Am Soc Nephrol 5:201–209Google Scholar
  3. Bachmann S, Le Hir M, Eckardt KU (1993) Co-localization of erythropoietin mRNA and ecto-5′-nucleotidase immunoreactivity in peritubular cells of rat renal cortex indicates that fibroblasts produce erythropoietin. J Histochem Cytochem 41:335–341Google Scholar
  4. Barraclough R (1998) Calcium-binding protein S100A4 in health and disease. Biochim Biophys Acta 1448:190–199Google Scholar
  5. Basile DP, Fredrich K, Alausa M, Vio CP, Liang M, Rieder MR, Green AS, Cowley AW Jr (2005) Identification of persistently altered gene expression in kidney following functional recovery from ischemic renal failure. Am J Physiol Renal Physiol 288:F953–F963Google Scholar
  6. Bhowmick NA, Chytil A, Plieth D, Gorska AE, Dumont N, Shappell S, Washington MK, Neilson EG, Moses HL (2004) TGF-beta signaling in fibroblasts modulates the oncogenic potential of adjacent epithelia. Science 303:848–851Google Scholar
  7. Bulger RE, Nagle RB (1973) Ultrastructure of the interstitium in the rabbit kidney. Am J Anat 136:183–203Google Scholar
  8. Chai Q, Krag S, Chai S, Ledet T, Wogensen L (2003) Localisation and phenotypical characterisation of collagen-producing cells in TGF-beta 1-induced renal interstitial fibrosis. Histochem Cell Biol 119:267–280Google Scholar
  9. Dawson TP, Gandhi R, Le Hir M, Kaissling B (1989) Ecto 1-5′-nucleotidase: localization in rat kidney by light microscopic histochemical and immunohistochemical methods. J Histochem Cytochem 37:39–47Google Scholar
  10. Desmouliere A, Darby IA, Gabbiani G (2003) Normal and pathologic soft tissue remodeling: role of the myofibroblast, with special emphasis on liver and kidney fibrosis. Lab Invest 83:1689–1707Google Scholar
  11. Diamond JR, van Goor H, Ding G, Engelmyer E (1995) Myofibroblasts in experimental hydronephrosis. Am J Pathol 146:121–129Google Scholar
  12. Duarte WR, Shibata T, Takenaga K, Takahashi E, Kubota K, Ohya K, Ishikawa I, Yamauchi M, Kasugai S (2003) S100A4: a novel negative regulator of mineralization and osteoblast differentiation. J Bone Miner Res 18:493–501Google Scholar
  13. El-Nahas AM (2003) Plasticity of kidney cells: role in kidney remodeling and scarring. Kidney Int 64:1553–1563Google Scholar
  14. Fearns C, Kravchenko VV, Ulevitch RJ, Loskutoff DJ (1995) Murine CD14 gene expression in vivo: extramyeloid synthesis and regulation by lipopolysaccharide. J Exp Med 181:857–866Google Scholar
  15. Gandhi R, Le Hir M, Kaissling B (1990) Immunolocalization of ecto-5′-nucleotidase in the kidney by a monoclonal antibody. Histochemistry 95:165–174Google Scholar
  16. Haeryfar SMM, Hoskin DW (2004) Thy-1: More than a mouse pan-T cell marker. J Immunol 173:3581–3588Google Scholar
  17. Herzlinger D (2002) Renal interstitial fibrosis: remembrance of things past? J Clin Invest 110:305–306Google Scholar
  18. Hong Y, Zhou W, Li K, Sacks SH (2002) Triptolide is a potent suppressant of C3, CD40 and B7h expression in activated human proximal tubular epithelial cells. Kidney Int 62:1291–1300Google Scholar
  19. Ito K, Chen J, El Chaar M, Stern JM, Seshan SV, Khodadadian JJ, Richardson I, Hyman MJ, Vaughan ED Jr, Poppas DP, Felsen D (2004) Renal damage progresses despite improvement of renal function after relief of unilateral ureteral obstruction in adult rats. Am J Physiol Renal Physiol 287:F1283–F1293Google Scholar
  20. Iwano M, Neilson EG (2004) Mechanisms of tubulointerstitial fibrosis. Curr Opin Nephrol Hypertens 13:279–284Google Scholar
  21. Iwano M, Plieth D, Danoff TM, Xue C, Okada H, Neilson EG (2002) Evidence that fibroblasts derive from epithelium during tissue fibrosis. J Clin Invest 110:341–350Google Scholar
  22. Jinde K, Nikolic-Paterson DJ, Huang XR, Sakai H, Kurokawa K, Atkins RC, Lan HY (2001) Tubular phenotypic change in progressive tubulointerstitial fibrosis in human glomerulonephritis. Am J Kidney Dis 38:761–769Google Scholar
  23. Kaissling B, Le Hir M (1994) Characterization and distribution of interstitial cell types in the renal cortex of rats. Kidney Int 45:709–720Google Scholar
  24. Kaissling B, Bachmann S, Kriz W (1985) Structural adaptation of the distal convoluted tubule to prolonged furosemide treatment. Am J Physiol 248:F374–F381Google Scholar
  25. Kaissling B, Hegyi I, Loffing J, Le Hir M (1996) Morphology of interstitial cells in the healthy kidney. Anat Embryol (Berl) 193:303–318Google Scholar
  26. Kalluri R, Neilson EG (2003) Epithelial–mesenchymal transition and its implications for fibrosis. J Clin Invest 112:1776–1784Google Scholar
  27. Kang Y, Massague J (2004) Epithelial–mesenchymal transitions: twist in development and metastasis. Cell 118:277–279Google Scholar
  28. van Kooten C, Woltman AM, Daha MR (2000) Immunological function of tubular epithelial cells: the functional implications of CD40 expression. Exp Nephrol 8:203–207Google Scholar
  29. Lawson WE, Polosukhin VV, Zoia O, Stathopoulos GT, Han W, Plieth D, Loyd JE, Neilson EG, Blackwell TS (2004) Characterization of fibroblast specific protein 1 in pulmonary fibrosis. Am J Respir Crit Care Med 23:23Google Scholar
  30. Le Hir M, Kaissling B (1989) Distribution of 5′-nucleotidase in the renal interstitium of the rat. Cell Tissue Res 258:177–182Google Scholar
  31. Le Hir M, Besse-Eschmann V (2003) A novel mechanism of nephron loss in a murine model of crescentic glomerulonephritis. Kidney Int 63:591–599Google Scholar
  32. Le Hir M, Keller C, Eschmann V, Hahnel B, Hosser H, Kriz W (2001) Podocyte bridges between the tuft and Bowman’s capsule: an early event in experimental crescentic glomerulonephritis. J Am Soc Nephrol 12:2060–2071Google Scholar
  33. Lemley KV, Kriz W (1991) Anatomy of the renal interstitium. Kidney Int 39:370–381Google Scholar
  34. Li Y, Yang J, Dai C, Wu C, Liu Y (2003) Role for integrin-linked kinase in mediating tubular epithelial to mesenchymal transition and renal interstitial fibrogenesis. J Clin Invest 112:503–516Google Scholar
  35. Liu Y (2004) Epithelial to mesenchymal transition in renal fibrogenesis: pathologic significance, molecular mechanism, and therapeutic intervention. J Am Soc Nephrol 15:1–12Google Scholar
  36. Loffing J, Loffing-Cueni D, Hegyi I, Kaplan MR, Hebert SC, Le Hir M, Kaissling B (1996) Thiazide treatment of rats provokes apoptosis in distal tubule cells. Kidney Int 50:1180–1190Google Scholar
  37. Marxer-Meier A, Hegyi I, Loffing J, Kaissling B (1998) Postnatal maturation of renal cortical peritubular fibroblasts in the rat. Anat Embryol (Berl) 197:143–153Google Scholar
  38. Masuda K, Masuda R, Neidhart M, Simmen BR, Michel BA, Muller-Ladner U, Gay RE, Gay S (2002) Molecular profile of synovial fibroblasts in rheumatoid arthritis depends on the stage of proliferation. Arthritis Res 4:R8Google Scholar
  39. Mazzetti I, Magagnoli G, Paoletti S, Uguccioni M, Olivotto E, Vitellozzi R, Cattini L, Facchini A, Borzi RM (2004) A role for chemokines in the induction of chondrocyte phenotype modulation. Arthritis Rheum 50:112–122Google Scholar
  40. Mazzucchelli L (2002) Protein S100A4: too long overlooked by pathologists? Am J Pathol 160:7–13Google Scholar
  41. Morrissey J, Guo G, McCracken R, Tolley T, Klahr S (2000) Induction of CD14 in tubular epithelial cells during kidney disease. J Am Soc Nephrol 11:1681–1690Google Scholar
  42. Ng YY, Huang TP, Yang WC, Chen ZP, Yang AH, Mu W, Nikolic-Paterson DJ, Atkins RC, Lan HY (1998) Tubular epithelial–myofibroblast transdifferentiation in progressive tubulointerstitial fibrosis in 5/6 nephrectomized rats. Kidney Int 54:864–876Google Scholar
  43. Niemann-Masanek U, Mueller A, Yard BA, Waldherr R, van der Woude FJ (2002) B7–1 (CD80) and B7–2 (CD 86) expression in human tubular epithelial cells in vivo and in vitro. Nephron 92:542–556Google Scholar
  44. Okada H, Ban S, Nagao S, Takahashi H, Suzuki H, Neilson EG (2000) Progressive renal fibrosis in murine polycystic kidney disease: an immunohistochemical observation. Kidney Int 58:587–597Google Scholar
  45. Okada H, Inoue T, Kanno Y, Kobayashi T, Watanabe Y, Ban S, Neilson EG, Suzuki H (2003) Selective depletion of fibroblasts preserves morphology and the functional integrity of peritoneum in transgenic mice with peritoneal fibrosing syndrome. Kidney Int 64:1722–1732Google Scholar
  46. Pfaller W (1982) Structure function correlation on rat kidney. Quantitative correlation of structure and function in the normal and injured rat kidney. Adv Anat Embryol Cell Biol 70:1–106Google Scholar
  47. Phan SH (2002) The myofibroblast in pulmonary fibrosis. Chest 122:286S–289SGoogle Scholar
  48. Pichler RH, Hugo C, Shankland SJ, Reed MJ, Bassuk JA, Andoh TF, Lombardi DM, Schwartz SM, Bennett WM, Alpers CE, Sage EH, Johnson RJ, Couser WG (1996) SPARC is expressed in renal interstitial fibrosis and in renal vascular injury. Kidney Int 50:1978–1989Google Scholar
  49. Reilly RF and Ellison DH (2000) Mammalian distal tubule: physiology, pathophysiology, and molecular anatomy. Physiol Rev 80:277–313Google Scholar
  50. Ryan DG, Taliana L, Sun L, Wei ZG, Masur SK, Lavker RM (2003) Involvement of S100A4 in stromal fibroblasts of the regenerating cornea. Invest Ophthalmol Vis Sci 44:4255–4262Google Scholar
  51. Sandelin M, Zabihi S, Liu L, Wicher G, Kozlova EN (2004) Metastasis-associated S100A4 (Mts1) protein is expressed in subpopulations of sensory and autonomic neurons and in Schwann cells of the adult rat. J Comp Neurol 473:233–243Google Scholar
  52. Sartore S, Chiavegato A, Faggin E, Franch R, Puato M, Ausoni S, Pauletto P (2001) Contribution of adventitial fibroblasts to neointima formation and vascular remodeling: from innocent bystander to active participant. Circ Res 89:1111–1121Google Scholar
  53. Short M, Nemenoff RA, Zawada WM, Stenmark KR, Das M (2004) Hypoxia induces differentiation of pulmonary artery adventitial fibroblasts into myofibroblasts. Am J Physiol Cell Physiol 286:C416–C425Google Scholar
  54. Spurgeon KS, Donohoe DL, Basile DP (2004) Transforming growth factor-beta in acute renal failure: receptor expression, effects on proliferation, cellularity and vascularization after recovery from injury. Am J Physiol Renal Physiol 9:9Google Scholar
  55. Strutz F, Neilson EG (2003) New insights into mechanisms of fibrosis in immune renal injury. Springer Semin Immunopathol 24:459–476Google Scholar
  56. Strutz F, Okada H, Lo CW, Danoff T, Carone RL, Tomaszewski JE, Neilson EG (1995) Identification and characterization of a fibroblast marker: FSP1. J Cell Biol 130:393–405Google Scholar
  57. Takei Y, Sims TN, Urmson J, Halloran PF (2000) Central role for interferon-gamma receptor in the regulation of renal MHC expression. J Am Soc Nephrol 11:250–261Google Scholar
  58. Taylor S, Herrington S, Prime W, Rudland PS, Barraclough R (2002) S100A4 (p9Ka) protein in colon carcinoma and liver metastases: association with carcinoma cells and T-lymphocytes. Br J Cancer 86:409–416Google Scholar
  59. Thiery JP (2002) Epithelial–mesenchymal transitions in tumour progression. Nat Rev Cancer 2:442–454Google Scholar
  60. Vielhauer V, Berning E, Eis V, Kretzler M, Segerer S, Strutz F, Horuk R, Grone HJ, Schlondorff D, Anders HJ (2004) CCR1 blockade reduces interstitial inflammation and fibrosis in mice with glomerulosclerosis and nephrotic syndrome. Kidney Int 66:2264–2278Google Scholar
  61. Witzgall R, Brown D, Schwarz C, Bonventre JV (1994) Localization of proliferating cell nuclear antigen, vimentin, c-Fos, and clusterin in the postischemic kidney. Evidence for a heterogenous genetic response among nephron segments, and a large pool of mitotically active and dedifferentiated cells. J Clin Invest 93:2175–2188Google Scholar
  62. Wuthrich RP (1992) Intercellular adhesion molecules and vascular cell adhesion molecule-1 and the kidney. J Am Soc Nephrol 3:1201–1211Google Scholar
  63. Yang J, Liu Y (2001) Dissection of key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis. Am J Pathol 159:1465–1475Google Scholar
  64. Yang J, Liu Y (2002) Blockage of tubular epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis. J Am Soc Nephrol 13:96–107Google Scholar
  65. Zeisberg M, Strutz F, Muller GA (2001) Renal fibrosis: an update. Curr Opin Nephrol Hypertens 10:315–320Google Scholar
  66. Zhu MQ, De Broe ME, Nouwen EJ (1996) Vimentin expression and distal tubular damage in the rat kidney. Exp Nephrol 4:172–183Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Michel Le Hir
    • 1
  • Ivan Hegyi
    • 2
  • Dominique Cueni-Loffing
    • 1
  • Johannes Loffing
    • 3
  • Brigitte Kaissling
    • 1
  1. 1.Anatomical Institute Division of Vegetative Anatomy, University of ZurichZurichSwitzerland
  2. 2.Department of PathologyInstitut für Klinische Pathologie, University of ZürichZurichSwitzerland
  3. 3.Institut d‘Anatomie Université de FribourgZurichSwitzerland

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