Laser capture microdissection/GeneChip analysis of gene expression in glomerular cells in diabetic db/db mice

Abstract

Although the gene expression patterns during the development of diabetic nephropathy have been studied in both rodent models and humans, only a small portion of the mRNAs expressed in the mesangium or in glomerular cells has been characterized. In the present study we report larger groups of transcripts displaying significant expression modulation in glomerular cells obtained from the early phase of diabetic nephropathy.

Methods: We used 12-week-old female db/db mice, a rodent model of type 2 diabetes, and their non-diabetic db/m litter-mates. Glomerular cells were obtained from the kidneys of mice by laser capture microdissection. Preparation of cRNA and target hybridization were performed according to the Affymetrix GeneChip Eukaryotic Small Sample Target Labeling Assay Protocol (Version II). The gene expression profile was determined by the mouse Expression Set 430A GeneChip.

Results: By comparison between db/m and db/db mice, 649 probes that increased in expression with the induction of diabetes and 340 probes that decreased in diabetic kidneys were identified. Although some of these genes have previously been shown to play an important role in diabetic nephropathy, the large majority of them have never been demonstrated to be regulated during the development of nephropathy.

Conclusions: Although the precise involvement of these genes in diabetic nephropathy remains to be clarified, the data presented here will aid in the identification of genes that play a significant role in this pathological condition.

Keywords:

• DB/DB MOUSE
• DNA MICROARRAY
• DIABETIC NEPHROPATHY
• GENE EXPRESSION
• LASER CAPTURE
• MICRODISSECTION