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. 1995 Jul 2;130(2):393–405. doi: 10.1083/jcb.130.2.393

Identification and characterization of a fibroblast marker: FSP1

PMCID: PMC2199940  PMID: 7615639

Abstract

We performed subtractive and differential hybridization for transcript comparison between murine fibroblasts and isogenic epithelium, and observed only a few novel intracellular genes which were relatively specific for fibroblasts. One such gene encodes a filament-associated, calcium-binding protein, fibroblast-specific protein 1 (FSP1). The promoter/enhancer region driving this gene is active in fibroblasts but not in epithelium, mesangial cells or embryonic endoderm. During development, FSP1 is first detected by in situ hybridization after day 8.5 as a postgastrulation event, and is associated with cells of mesenchymal origin or of fibroblastic phenotype. Polyclonal antiserum raised to recombinant FSP1 protein stained the cytoplasm of fibroblasts, but not epithelium. Only occasional cells stain with specific anti-FSP1 antibodies in normal parenchymal tissue. However, in kidneys fibrosing from persistent inflammation, many fibroblasts could be identified in interstitial sites of collagen deposition and also in tubular epithelium adjacent to the inflammatory process. This pattern of anti-FSP1 staining during tissue fibrosis suggests, as a hypothesis, that fibroblasts in some cases arise, as needed, from the local conversion of epithelium. Consistent with this notion that FSP1 may be involved in the transition from epithelium to fibroblasts are experiments in which the in vitro overexpression of FSP1 cDNA in tubular epithelium is accompanied by conversion to a mesenchymal phenotype, as characterized by a more stellate and elongated fibroblast- like appearance, a reduction in cytokeratin, and new expression of vimentin. Similarly, tubular epithelium submerged in type I collagen gels exhibited the conversion to a fibroblast phenotype which includes de novo expression of FSP1 and vimentin. Use of the FSP1 marker, therefore, should further facilitate both the in vivo studies of fibrogenesis and the mapping of cell fate among fibroblasts.

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Selected References

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  1. Abrahamsohn P. A., Zorn T. M. Implantation and decidualization in rodents. J Exp Zool. 1993 Sep 1;266(6):603–628. doi: 10.1002/jez.1402660610. [DOI] [PubMed] [Google Scholar]
  2. Agelli M., Wahl S. M. Cytokines and fibrosis. Clin Exp Rheumatol. 1986 Oct-Dec;4(4):379–388. [PubMed] [Google Scholar]
  3. Albert S. E., Strutz F., Shelton K., Haverty T., Sun M. J., Li S. R., Denham A., Maki R. A., Neilson E. G. Characterization of a cis-acting regulatory element which silences expression of the class II-A beta gene in epithelium. J Exp Med. 1994 Jul 1;180(1):233–240. doi: 10.1084/jem.180.1.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Alpers C. E., Seifert R. A., Hudkins K. L., Johnson R. J., Bowen-Pope D. F. PDGF-receptor localizes to mesangial, parietal epithelial, and interstitial cells in human and primate kidneys. Kidney Int. 1993 Feb;43(2):286–294. doi: 10.1038/ki.1993.45. [DOI] [PubMed] [Google Scholar]
  5. Alvarez R. J., Sun M. J., Haverty T. P., Iozzo R. V., Myers J. C., Neilson E. G. Biosynthetic and proliferative characteristics of tubulointerstitial fibroblasts probed with paracrine cytokines. Kidney Int. 1992 Jan;41(1):14–23. doi: 10.1038/ki.1992.3. [DOI] [PubMed] [Google Scholar]
  6. Anderson D. J., Axel R. Molecular probes for the development and plasticity of neural crest derivatives. Cell. 1985 Sep;42(2):649–662. doi: 10.1016/0092-8674(85)90122-9. [DOI] [PubMed] [Google Scholar]
  7. Bachem M. G., Meyer D., Melchior R., Sell K. M., Gressner A. M. Activation of rat liver perisinusoidal lipocytes by transforming growth factors derived from myofibroblastlike cells. A potential mechanism of self perpetuation in liver fibrogenesis. J Clin Invest. 1992 Jan;89(1):19–27. doi: 10.1172/JCI115561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Baudier J., Cole R. D. Interactions between the microtubule-associated tau proteins and S100b regulate tau phosphorylation by the Ca2+/calmodulin-dependent protein kinase II. J Biol Chem. 1988 Apr 25;263(12):5876–5883. [PubMed] [Google Scholar]
  9. Bornstein P., Sage H. Regulation of collagen gene expression. Prog Nucleic Acid Res Mol Biol. 1989;37:67–106. doi: 10.1016/s0079-6603(08)60695-9. [DOI] [PubMed] [Google Scholar]
  10. Bradley A., Evans M., Kaufman M. H., Robertson E. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature. 1984 May 17;309(5965):255–256. doi: 10.1038/309255a0. [DOI] [PubMed] [Google Scholar]
  11. Breen E., Falco V. M., Absher M., Cutroneo K. R. Subpopulations of rat lung fibroblasts with different amounts of type I and type III collagen mRNAs. J Biol Chem. 1990 Apr 15;265(11):6286–6290. [PubMed] [Google Scholar]
  12. Broekelmann T. J., Limper A. H., Colby T. V., McDonald J. A. Transforming growth factor beta 1 is present at sites of extracellular matrix gene expression in human pulmonary fibrosis. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6642–6646. doi: 10.1073/pnas.88.15.6642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Butler W. T. The nature and significance of osteopontin. Connect Tissue Res. 1989;23(2-3):123–136. doi: 10.3109/03008208909002412. [DOI] [PubMed] [Google Scholar]
  14. Danoff T. M., Lalley P. A., Chang Y. S., Heeger P. S., Neilson E. G. Cloning, genomic organization, and chromosomal localization of the Scya5 gene encoding the murine chemokine RANTES. J Immunol. 1994 Feb 1;152(3):1182–1189. [PubMed] [Google Scholar]
  15. Davies B. R., Davies M. P., Gibbs F. E., Barraclough R., Rudland P. S. Induction of the metastatic phenotype by transfection of a benign rat mammary epithelial cell line with the gene for p9Ka, a rat calcium-binding protein, but not with the oncogene EJ-ras-1. Oncogene. 1993 Apr;8(4):999–1008. [PubMed] [Google Scholar]
  16. Dawson T. P., Gandhi R., Le Hir M., Kaissling B. Ecto-5'-nucleotidase: localization in rat kidney by light microscopic histochemical and immunohistochemical methods. J Histochem Cytochem. 1989 Jan;37(1):39–47. doi: 10.1177/37.1.2535703. [DOI] [PubMed] [Google Scholar]
  17. Dony C., Gruss P. Specific expression of the Hox 1.3 homeo box gene in murine embryonic structures originating from or induced by the mesoderm. EMBO J. 1987 Oct;6(10):2965–2975. doi: 10.1002/j.1460-2075.1987.tb02602.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ebralidze A., Tulchinsky E., Grigorian M., Afanasyeva A., Senin V., Revazova E., Lukanidin E. Isolation and characterization of a gene specifically expressed in different metastatic cells and whose deduced gene product has a high degree of homology to a Ca2+-binding protein family. Genes Dev. 1989 Jul;3(7):1086–1093. doi: 10.1101/gad.3.7.1086. [DOI] [PubMed] [Google Scholar]
  19. Ekblom P., Lehtonen E., Saxén L., Timpl R. Shift in collagen type as an early response to induction of the metanephric mesenchyme. J Cell Biol. 1981 May;89(2):276–283. doi: 10.1083/jcb.89.2.276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Enders A. C. Current topic: structural responses of the primate endometrium to implantation. Placenta. 1991 Jul-Aug;12(4):309–325. doi: 10.1016/0143-4004(91)90340-l. [DOI] [PubMed] [Google Scholar]
  21. French B. T., Lee W. H., Maul G. G. Nucleotide sequence of a cDNA clone for mouse pro alpha 1(I) collagen protein. Gene. 1985;39(2-3):311–312. doi: 10.1016/0378-1119(85)90329-4. [DOI] [PubMed] [Google Scholar]
  22. Friedman S. L., Arthur M. J. Activation of cultured rat hepatic lipocytes by Kupffer cell conditioned medium. Direct enhancement of matrix synthesis and stimulation of cell proliferation via induction of platelet-derived growth factor receptors. J Clin Invest. 1989 Dec;84(6):1780–1785. doi: 10.1172/JCI114362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Friedman S. L. Seminars in medicine of the Beth Israel Hospital, Boston. The cellular basis of hepatic fibrosis. Mechanisms and treatment strategies. N Engl J Med. 1993 Jun 24;328(25):1828–1835. doi: 10.1056/NEJM199306243282508. [DOI] [PubMed] [Google Scholar]
  24. Frixen U. H., Behrens J., Sachs M., Eberle G., Voss B., Warda A., Löchner D., Birchmeier W. E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells. J Cell Biol. 1991 Apr;113(1):173–185. doi: 10.1083/jcb.113.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Gilligan A., Jewett L., Simon D., Damjanov I., Matschinsky F. M., Weik H., Pinkert C., Knowles B. B. Functional pancreatic beta-cell line from SV40 T-antigen transgenic mouse. Diabetes. 1989 Aug;38(8):1056–1062. doi: 10.2337/diab.38.8.1056. [DOI] [PubMed] [Google Scholar]
  26. Goldberg H., Helaakoski T., Garrett L. A., Karsenty G., Pellegrino A., Lozano G., Maity S., de Crombrugghe B. Tissue-specific expression of the mouse alpha 2(I) collagen promoter. Studies in transgenic mice and in tissue culture cells. J Biol Chem. 1992 Sep 25;267(27):19622–19630. [PubMed] [Google Scholar]
  27. Goto K., Endo H., Fujiyoshi T. Cloning of the sequences expressed abundantly in established cell lines: identification of a cDNA clone highly homologous to S-100, a calcium binding protein. J Biochem. 1988 Jan;103(1):48–53. doi: 10.1093/oxfordjournals.jbchem.a122237. [DOI] [PubMed] [Google Scholar]
  28. Greenburg G., Hay E. D. Cytodifferentiation and tissue phenotype change during transformation of embryonic lens epithelium to mesenchyme-like cells in vitro. Dev Biol. 1986 Jun;115(2):363–379. doi: 10.1016/0012-1606(86)90256-3. [DOI] [PubMed] [Google Scholar]
  29. Greenburg G., Hay E. D. Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells. J Cell Biol. 1982 Oct;95(1):333–339. doi: 10.1083/jcb.95.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Hagiwara M., Ochiai M., Owada K., Tanaka T., Hidaka H. Modulation of tyrosine phosphorylation of p36 and other substrates by the S-100 protein. J Biol Chem. 1988 May 5;263(13):6438–6441. [PubMed] [Google Scholar]
  31. Haverty T. P., Kelly C. J., Hoyer J. R., Alvarez R., Neilson E. G. Tubular antigen-binding proteins repress transcription of type IV collagen in the autoimmune target epithelium of experimental interstitial nephritis. J Clin Invest. 1992 Feb;89(2):517–523. doi: 10.1172/JCI115615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  33. Iozzo R. V. Proteoglycans: structure, function, and role in neoplasia. Lab Invest. 1985 Oct;53(4):373–396. [PubMed] [Google Scholar]
  34. Iwai N., Inagami T. Isolation of preferentially expressed genes in the kidneys of hypertensive rats. Hypertension. 1991 Feb;17(2):161–169. doi: 10.1161/01.hyp.17.2.161. [DOI] [PubMed] [Google Scholar]
  35. Jackson-Grusby L. L., Swiergiel J., Linzer D. I. A growth-related mRNA in cultured mouse cells encodes a placental calcium binding protein. Nucleic Acids Res. 1987 Aug 25;15(16):6677–6690. doi: 10.1093/nar/15.16.6677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Komuro T. Re-evaluation of fibroblasts and fibroblast-like cells. Anat Embryol (Berl) 1990;182(2):103–112. doi: 10.1007/BF00174011. [DOI] [PubMed] [Google Scholar]
  37. Kuncio G. S., Neilson E. G., Haverty T. Mechanisms of tubulointerstitial fibrosis. Kidney Int. 1991 Mar;39(3):550–556. doi: 10.1038/ki.1991.63. [DOI] [PubMed] [Google Scholar]
  38. Kähäri V. M., Sandberg M., Kalimo H., Vuorio T., Vuorio E. Identification of fibroblasts responsible for increased collagen production in localized scleroderma by in situ hybridization. J Invest Dermatol. 1988 May;90(5):664–670. doi: 10.1111/1523-1747.ep12560826. [DOI] [PubMed] [Google Scholar]
  39. Lakshmi M. S., Parker C., Sherbet G. V. Metastasis associated MTS1 and NM23 genes affect tubulin polymerisation in B16 melanomas: a possible mechanism of their regulation of metastatic behaviour of tumours. Anticancer Res. 1993 Mar-Apr;13(2):299–303. [PubMed] [Google Scholar]
  40. Lamb B. T., Satyamoorthy K., Solter D., Basu A., Xu M. Q., Weinmann R., Howe C. C. A DNA element that regulates expression of an endogenous retrovirus during F9 cell differentiation is E1A dependent. Mol Cell Biol. 1992 Nov;12(11):4824–4833. doi: 10.1128/mcb.12.11.4824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Le Hir M., Kaissling B. Distribution of 5'-nucleotidase in the renal interstitium of the rat. Cell Tissue Res. 1989 Oct;258(1):177–182. doi: 10.1007/BF00223156. [DOI] [PubMed] [Google Scholar]
  42. Liau G., Yamada Y., de Crombrugghe B. Coordinate regulation of the levels of type III and type I collagen mRNA in most but not all mouse fibroblasts. J Biol Chem. 1985 Jan 10;260(1):531–536. [PubMed] [Google Scholar]
  43. Linzer D. I., Nathans D. Growth-related changes in specific mRNAs of cultured mouse cells. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4271–4275. doi: 10.1073/pnas.80.14.4271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Lopez C. A., Hoyer J. R., Wilson P. D., Waterhouse P., Denhardt D. T. Heterogeneity of osteopontin expression among nephrons in mouse kidneys and enhanced expression in sclerotic glomeruli. Lab Invest. 1993 Sep;69(3):355–363. [PubMed] [Google Scholar]
  45. Maher J. J., McGuire R. F. Extracellular matrix gene expression increases preferentially in rat lipocytes and sinusoidal endothelial cells during hepatic fibrosis in vivo. J Clin Invest. 1990 Nov;86(5):1641–1648. doi: 10.1172/JCI114886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Mann R., Kelly C. J., Hines W. H., Clayman M. D., Blanchard N., Sun M. J., Neilson E. G. Effector T cell differentiation in experimental interstitial nephritis. I. The development and modulation of effector lymphocyte maturation by I-J+ regulatory T cells. J Immunol. 1987 Jun 15;138(12):4200–4208. [PubMed] [Google Scholar]
  47. Merritt S. E., Killen P. D., Phan S. H., Wiggins R. C. Analysis of alpha 1 (I) procollagen alpha 1 (IV) collagen, and beta-actin mRNA in glomerulus and cortex of rabbits with experimental anti-glomerular basement membrane disease. Evidence for early extraglomerular collagen biosynthesis. Lab Invest. 1990 Dec;63(6):762–769. [PubMed] [Google Scholar]
  48. Miyazaki Y., Setoguchi M., Yoshida S., Higuchi Y., Akizuki S., Yamamoto S. The mouse osteopontin gene. Expression in monocytic lineages and complete nucleotide sequence. J Biol Chem. 1990 Aug 25;265(24):14432–14438. [PubMed] [Google Scholar]
  49. Nakatsukasa H., Nagy P., Evarts R. P., Hsia C. C., Marsden E., Thorgeirsson S. S. Cellular distribution of transforming growth factor-beta 1 and procollagen types I, III, and IV transcripts in carbon tetrachloride-induced rat liver fibrosis. J Clin Invest. 1990 Jun;85(6):1833–1843. doi: 10.1172/JCI114643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Neilson E. G., Kalluri R., Sun M. J., Gunwar S., Danoff T., Mariyama M., Myers J. C., Reeders S. T., Hudson B. G. Specificity of Goodpasture autoantibodies for the recombinant noncollagenous domains of human type IV collagen. J Biol Chem. 1993 Apr 25;268(12):8402–8405. [PubMed] [Google Scholar]
  51. Nordeen S. K. Luciferase reporter gene vectors for analysis of promoters and enhancers. Biotechniques. 1988 May;6(5):454–458. [PubMed] [Google Scholar]
  52. Obeso J., Weber J., Auerbach R. A hemangioendothelioma-derived cell line: its use as a model for the study of endothelial cell biology. Lab Invest. 1990 Aug;63(2):259–269. [PubMed] [Google Scholar]
  53. Parry D. A. The molecular and fibrillar structure of collagen and its relationship to the mechanical properties of connective tissue. Biophys Chem. 1988 Feb;29(1-2):195–209. doi: 10.1016/0301-4622(88)87039-x. [DOI] [PubMed] [Google Scholar]
  54. Phillips C. L., Lever L. W., Pinnell S. R., Quarles L. D., Wenstrup R. J. Construction of a full-length murine pro alpha 2(I) collagen cDNA by the polymerase chain reaction. J Invest Dermatol. 1991 Dec;97(6):980–984. doi: 10.1111/1523-1747.ep12491894. [DOI] [PubMed] [Google Scholar]
  55. Rosenthal N. Identification of regulatory elements of cloned genes with functional assays. Methods Enzymol. 1987;152:704–720. doi: 10.1016/0076-6879(87)52075-4. [DOI] [PubMed] [Google Scholar]
  56. Sappino A. P., Schürch W., Gabbiani G. Differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab Invest. 1990 Aug;63(2):144–161. [PubMed] [Google Scholar]
  57. Schneppenheim R., Budde U., Dahlmann N., Rautenberg P. Luminography--a new, highly sensitive visualization method for electrophoresis. Electrophoresis. 1991 May;12(5):367–372. doi: 10.1002/elps.1150120508. [DOI] [PubMed] [Google Scholar]
  58. Schwartz A. L., Fridovich S. E., Knowles B. B., Lodish H. F. Characterization of the asialoglycoprotein receptor in a continuous hepatoma line. J Biol Chem. 1981 Sep 10;256(17):8878–8881. [PubMed] [Google Scholar]
  59. Shanahan C. M., Weissberg P. L., Metcalfe J. C. Isolation of gene markers of differentiated and proliferating vascular smooth muscle cells. Circ Res. 1993 Jul;73(1):193–204. doi: 10.1161/01.res.73.1.193. [DOI] [PubMed] [Google Scholar]
  60. Sive H. L., St John T. A simple subtractive hybridization technique employing photoactivatable biotin and phenol extraction. Nucleic Acids Res. 1988 Nov 25;16(22):10937–10937. doi: 10.1093/nar/16.22.10937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Somerman M. J., Prince C. W., Sauk J. J., Foster R. A., Butler W. T. Mechanism of fibroblast attachment to bone extracellular matrix: role of a 44 kilodalton bone phosphoprotein. J Bone Miner Res. 1987 Jun;2(3):259–265. doi: 10.1002/jbmr.5650020313. [DOI] [PubMed] [Google Scholar]
  62. Sommers C. L., Heckford S. E., Skerker J. M., Worland P., Torri J. A., Thompson E. W., Byers S. W., Gelmann E. P. Loss of epithelial markers and acquisition of vimentin expression in adriamycin- and vinblastine-resistant human breast cancer cell lines. Cancer Res. 1992 Oct 1;52(19):5190–5197. [PubMed] [Google Scholar]
  63. Sommers C. L., Walker-Jones D., Heckford S. E., Worland P., Valverius E., Clark R., McCormick F., Stampfer M., Abularach S., Gelmann E. P. Vimentin rather than keratin expression in some hormone-independent breast cancer cell lines and in oncogene-transformed mammary epithelial cells. Cancer Res. 1989 Aug 1;49(15):4258–4263. [PubMed] [Google Scholar]
  64. Strickland S., Smith K. K., Marotti K. R. Hormonal induction of differentiation in teratocarcinoma stem cells: generation of parietal endoderm by retinoic acid and dibutyryl cAMP. Cell. 1980 Sep;21(2):347–355. doi: 10.1016/0092-8674(80)90471-7. [DOI] [PubMed] [Google Scholar]
  65. Takenaga K., Nakamura Y., Endo H., Sakiyama S. Involvement of S100-related calcium-binding protein pEL98 (or mts1) in cell motility and tumor cell invasion. Jpn J Cancer Res. 1994 Aug;85(8):831–839. doi: 10.1111/j.1349-7006.1994.tb02955.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Watanabe Y., Usada N., Minami H., Morita T., Tsugane S., Ishikawa R., Kohama K., Tomida Y., Hidaka H. Calvasculin, as a factor affecting the microfilament assemblies in rat fibroblasts transfected by src gene. FEBS Lett. 1993 Jun 7;324(1):51–55. doi: 10.1016/0014-5793(93)81530-d. [DOI] [PubMed] [Google Scholar]
  67. Weber K. T. Cardiac interstitium in health and disease: the fibrillar collagen network. J Am Coll Cardiol. 1989 Jun;13(7):1637–1652. doi: 10.1016/0735-1097(89)90360-4. [DOI] [PubMed] [Google Scholar]
  68. Wiggins R., Goyal M., Merritt S., Killen P. D. Vascular adventitial cell expression of collagen I messenger ribonucleic acid in anti-glomerular basement membrane antibody-induced crescentic nephritis in the rabbit. A cellular source for interstitial collagen synthesis in inflammatory renal disease. Lab Invest. 1993 May;68(5):557–565. [PubMed] [Google Scholar]
  69. Wolf G., Haberstroh U., Neilson E. G. Angiotensin II stimulates the proliferation and biosynthesis of type I collagen in cultured murine mesangial cells. Am J Pathol. 1992 Jan;140(1):95–107. [PMC free article] [PubMed] [Google Scholar]

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