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      Dissociated Expression of Collagen Type IV Subchains in Diabetic Kidneys of KKAy Mice

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          Diabetic nephropathy is characterized by thickening of the glomerular basement membrane and expansion of the mesangial matrix. The glomerular basement membrane is assembled from at least five genetically distinct collagen IV chains. In patients with diabetic nephropathy, differential distribution of these components has been demonstrated. In order to clarify the relationship between progression of diabetic nephropathy and altered type IV collagen assembly in the renal cortex, we examined steady state mRNA levels encoding collagen IV subchains in the kidney cortices of spontaneously diabetic KKAy mice and nondiabetic C57black mice as controls. They were sacrificed at 4, 8, 16, and 24 weeks of age. Northern and dot blot analyses were performed using <sup>32</sup>P-labeled mouse probes for classical α1(IV) and α2(IV) and for α3(IV), α4(IV), and α5(IV) minor chains. The mRNA levels for all collagen IV chains peaked at 4 weeks of age and declined rapidly thereafter in the nondiabetic mice. At all times, α1(IV) and α2(IV) mRNA expressions were abundant and almost unchanged in KKAy mice. In contrast, mRNA levels for α3(IV), α4(IV), and α5(IV) progressively changed with age. It appears that the expression of minor collagen IV chains is dissociated from the α1(IV) and α2(IV) chains in diabetic nephropathy. Moreover, an unbalanced increase in the production may affect collagen IV assembly and contribute to basement membrane thickening in diabetic nephropathy of KKAy mice.

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.

            Author and article information

            S. Karger AG
            October 1998
            23 September 1998
            : 80
            : 2
            : 208-213
            Division of Nephrology, Department of Medicine, Juntendo University School of Medicine, Tokyo, Japan
            45169 Nephron 1998;80:208–213
            © 1998 S. Karger AG, Basel

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            Page count
            Figures: 5, Tables: 1, References: 28, Pages: 6
            Self URI (application/pdf): https://www.karger.com/Article/Pdf/45169
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