PMID: 7540573Jul 1, 1995Paper

Cloning and expression of cytokine-inducible nitric oxide synthase cDNA from rat islets of Langerhans

Diabetes
A E KarlsenT Mandrup-Poulsen

Abstract

An inducible nitric oxide (NO) synthase isoform (iNOS) is specifically induced in the beta-cells of interleukin (IL)-1 beta-exposed rat islets, suggesting a role for NO in the pathogenesis of type I diabetes. The aim of this study was to clone and characterize iNOS cDNA from cytokine-exposed islets. Neither NO production nor iNOS transcription could be detected in rat islets or in rat insulinoma RIN-5AH beta-cells cultured in the absence of cytokines. Addition of IL-1 beta alone or in combination with tumor necrosis factor-alpha induced a concentration- and time-dependent expression of the iNOS gene and associated NO production (measured as nitrite) from both islets and RIN cells. iNOS transcripts were cloned by reverse transcriptase-polymerase chain reaction from the cytokine-exposed rat islets and RIN cells, and DNA sequence analysis revealed a near 100% identity to the recently published iNOS cDNA cloned from cytokine-exposed rat hepatocytes and smooth muscle cells. Recombinant rat islet iNOS was transiently and stably expressed in human kidney 293 fibroblasts, and the high enzymatic activity was inhibited by addition of the L-arginine analogs, N omega-nitro-L-arginine methyl ester and aminoguanidine. Two-dimensional gel ele...Continue Reading

Citations

Nov 27, 2004·Diabetes Research and Clinical Practice·Eiji KawasakiKatsumi Eguchi
May 12, 2007·The Review of Diabetic Studies : RDS·Wageh M AwaraKarima el-Desoky
Jun 23, 2005·Diabetologia·C MathieuR Bouillon
Jun 23, 2011·Cellular Signalling·Kewei WangAi-Xuan Holterman
Sep 1, 1996·Diabetologia·T Mandrup-Poulsen
Jan 5, 2000·Journal of the American Society of Nephrology : JASN·Roland VeelkenR Bernd Sterzel
May 18, 2005·Biochemical Pharmacology·Kyoichiro MaeshimaKiyoshi Morita

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