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Localisation and phenotypical characterisation of collagen-producing cells in TGF-β1-induced renal interstitial fibrosis

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Abstract

Transforming growth factor β1 (TGF-β1) contributes to the accumulation of extracellular matrix (ECM) in the tubulointerstitial space in chronic renal diseases. Identification of target cells and the contribution of epithelial–mesenchymal transformation (EMT) in TGF-β1-induced fibrosis in vivo are currently under investigation. We have developed a transgenic model of slowly developing TGF-β1-driven tubulointerstitial fibrosis (TIF). By using this model our aim was to localise the ECM-producing cells, to investigate the temporal and spatial distribution of the cellular markers α-smooth muscle cell actin (αSM-actin), Fsp1 and Hsp47 and to explore the possible involvement of EMT in TGF-β1-induced TIF in vivo. We utilised a combination of in situ hybridisation, immunohistochemistry and western blotting techniques and found that αSM-actin-positive interstitial cells are the main source of collagen types I and III and fibronectin, whereas collagen type IV(α1/α2) originates mainly from the tubular epithelial cells. Furthermore, macrophages are not important combatants during the early course of TGF-β1-induced TIF. Finally, EMT is not necessary for the initiation of TGF-β1-induced TIF. We conclude, that intervention directed against the recruitment of activated interstitial cells may avoid the development of end-stage renal disease.

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Acknowledgements

We are indebted to Anita Morell, Karin Vestergaard and Mette Degn for their technical assistance. We would like to thank the helpful staff at the animal facility, The Bartholin Building, Aarhus University, Denmark. Finally, we would like to thank the following persons for their generous gifts of antibodies or probes: R. Hynes (MIT, Cambridge, Mass., USA), H. Eberspaecher (Anderson Cancer Center, The University of Texas, USA), L. Wood (Genomics Research, Pharmacia and Upjohn, Kalamazoo, Mich., USA) and E. Neilson (Vanderbilt University, Nashville, Tenn., USA). Søren Krag was the recipient of a scholarship from the Danish Medical Research Council (number 9701174). The work was supported by the Danish Medical Research Council (numbers 9601759, 9802621 and 9600822; Aarhus University Novo Nordisk Center for Research in Growth and Regeneration), the Novo Nordisk Foundation and The Institute of Experimental Clinical Research, Aarhus University, Aarhus.

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Correspondence to Lise Wogensen.

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Chai, Q., Krag, S., Chai, S. et al. Localisation and phenotypical characterisation of collagen-producing cells in TGF-β1-induced renal interstitial fibrosis. Histochem Cell Biol 119, 267–280 (2003). https://doi.org/10.1007/s00418-003-0513-8

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