Subtractive hybridization cloning: An efficient technique to detect overexpressed mRNAs in diabetic nephropathy Technical Note

Abstract

Hyperfiltration, associated with kidney hypertrophy and hyperplasia, are early markers of diabetic nephropathy [1, 2]. Procedures for the identification of renal molecules potentially involved in this early process are of particular interest. In order to detect hyperexpression of major renal proteins in early diabetic nephropathy, we used the well-known model of streptozotocin (STZ)induced diabetes in rats [3] and a robust technique of subtracted hybridization between normal and diabetic kidneys [4 ‐ 6]. Two subtracted probes were elaborated after hydridization of diabetic rat kidney cDNA with control rat kidney mRNA. These probes were used to screen a diabetic rat kidney cDNA library. Two clones were overexpressed in diabetic rat kidneys and corresponded to the multi-drug resistance 1 (mdr 1) cDNA and to the b-amyloid protein precursor (bPP), respectively. This study describes the results of subtractive hybridization cloning in rats, as well as some data obtained in human kidney sections originating from diabetic patients.