Nephrol Dial Transplant (2001) 16: 923-931
© 2001 European Renal Association-European Dialysis and Transplant Association
Rapid and diverse changes of gene expression in the kidneys of protein-overload proteinuria mice detected by microarray analysis
Department of Internal Medicine and Therapeutics, Graduate School of Medicine (A8), Osaka University, Osaka, Japan
Background. Microarray is a method that allows the analysis of a large number of genes at the same time. We applied this method to show the difference of gene expression in the kidney caused by proteinuria.
Methods. An experimental mouse model of protein overload was prepared by bovine serum albumin injection. The mRNAs of kidneys isolated after 0, 1, 2, 3 and 4 weeks loading were analysed by Northern blotting. We analysed about 18000 genes by microarray. The expression patterns of the microarray were displayed on control, 1 and 3 weeks of protein overload using the clustering procedure. A clone showing the greatest changes of up-regulation in the kidney was cloned and analysed by in situ hybridization and immunohistochemistry.
Results. Over 1600 kinds of gene expression were confirmed in control kidneys. Proteinuria caused systematic changes of gene expression demonstrated by the cluster analysis. The up-regulation of osteopontin mRNA was shown and confirmed by Northern blot analysis. One of the clones showing the largest changes, AA275245, was isolated and characterized. It revealed that AA275245 was an unreported 3' non-coding region of vinculin mRNA which was associated with cytoskeleton proteins (e.g. 
-actinin, talin, F-actin). Immunohistochemistry and in situ hybridization showed that this clone was identified in glomeruli as a mesangial pattern. The detected signal intensity using both methods, however, was virtually identical in control and disease kidney models. All data including images and analysed signal intensities are accessible on the web site.
Conclusion. The microarray analysis revealed that the renal gene expression pattern was changed dynamically in mice with experimentally induced proteinuria within a few weeks.
Keywords: gene expression; kidney; microarray; protein-overload; proteinuria; nephron segment
Correspondence and offprint requests to: Masaru Takenaka MD, Department of Internal Medicine and Therapeutics, Graduate School of Medicine (A8), Osaka University, 2–2 Yamadaoka, Suita, Osaka 560-0871, Japan.
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