NDT Advance Access originally published online on July 19, 2007
Nephrology Dialysis Transplantation 2007 22(10):3055-3060; doi:10.1093/ndt/gfm298
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A pitfall of glomerular sieving: profibrotic and matrix proteins derive from the Bowman's capsule and not the glomerular tuft in rats with renovascular hypertension
1Department of Medicine, Division of Nephrology and 2Center for Molecular Neurobiology, Department of Electron Microscopy, University Hospital Hamburg-Eppendorf, Germany
Correspondence and offprint requests to: Ulrich O. Wenzel, University Hospital of Hamburg-Eppendorf, Department of Medicine, Division of Nephrology, Martinistr. 52 20246 Hamburg, Germany. Email: wenzel{at}uke.uni-hamburg.de
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Abstract |
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Background. The glomeruli in the non-clipped kidney of rats with 2-kidney, 1-clip hypertension are a classical model for studying the mechanisms of glomerular injury.
Methods. In the present study, we compared the glomerular expression of PAI-1 and collagen I 
1 mRNA from glomeruli isolated by the classic technique of sieving with the recently developed technique of tissue laser microdissection. For quantification of mRNA from both methods, real-time PCR was used.
Results. Real-time PCR revealed a 9.0 ± 1.3- and a 7.1 ± 0.2-fold induction of PAI-1 and collagen I 1, respectively, in the glomeruli from hypertensive rats isolated by sieving. However, in situ hybridization and microdissection revealed that expression of both mRNAs was mainly from the Bowman's capsule and not from the glomerular tuft (10.7 ± 1.3- and 7.2 ± 0.6-fold higher induction in whole glomeruli compared with tuft alone).
Conclusion. This emphasizes that studies focusing on processes in the mesangium, endothelial cells or podocytes should not rely on glomeruli obtained by sieving. Rather, a technique like the laser microdissection or in situ hybridization should be applied which allows the clear separation of different glomerular and periglomerular compartments.
Keywords: Bowman's capsule; fibrosis; glomerular sieving; glomerular tuft; microdissection; PAI-1
See http://www.oxfordjournals.org/our_journals/ndtplus/
Received for publication: 16. 9.06
Accepted in revised form: 19. 4.07
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