2083 DIFFERENTIAL GENE EXPRESSION IN RAT KIDNEYS IN RESPONSE TO OXALATE AND CALCIUM OXALATE CRYSTALS: A TRANSCRIPTIONAL STUDY (2013)

You have accessJournal of UrologyStone Disease: Basic Research (I)1 Apr 20132083 DIFFERENTIAL GENE EXPRESSION IN RAT KIDNEYS IN RESPONSE TO OXALATE AND CALCIUM OXALATE CRYSTALS: A TRANSCRIPTIONAL STUDY Sunil Joshi, Wei Wang, Ammon Peck, and Saeed Khan Sunil JoshiSunil Joshi Gainesville, FL More articles by this author , Wei WangWei Wang Gainesville, FL More articles by this author , Ammon PeckAmmon Peck Gainesville, FL More articles by this author , and Saeed KhanSaeed Khan Gainesville, FL More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.2502AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Hyperoxaluria is responsible for a number of urological disorders. We have been investigating renal responses to oxalate ions (Ox) and calcium oxalate (CaOx) crystals in order to better understand the damaging consequences of hyperoxaluria. A variety of biological processes, molecular interactions, and signaling pathways are involved. In this study we used an unbiased approach of gene expression profiling to determine differential gene expression in the cortex and medulla of hyperoxaluric rat kidneys with and without crystals. METHODS Oxalate and calcium oxalate crystals were induced with hydroxy-l-proline in the kidneys of eight week old male Sprague Dawley rats. Urine was collected at day 7, 14, 21 and 28 and urinary oxalate, calcium, hydrogen peroxide, and citrate were measured. Rats were euthanized on day 28 and kidneys were extracted. Total RNA was isolated and micro array analysis was conducted for the crystals and no crystal groups using the Illumina bead array reader. Gene ontology (GO) analysis and the pathway analysis of the genes was done using DAVID (Database for Annotation, Visualization of Integrated Discovery) enrichment analysis tool. Western blot analysis and ELISA of urine samples were done to see the difference between crystals and no crystals groups. Statistical comparisons between the two groups were made by means of One way or Two way Analysis of Variance (ANOVA). RESULTS We examined kidneys of 6 rats, 5 controls, 3 with few or no crystals and 3 with heavy crystals deposition. From a total of 22,226 genes analyzed using the arrays, 2344 and 2948 genes were significantly differentially expressed in the cortex and medulla respectively in the no crystal group where as 3984 and 3949 genes were differentially expressed in the crystal group in cortex and medulla respectively. In the cortex and medulla there were 38 and 35 unique pathways between the no crystal and the crystal group respectively. Urinary oxalate and hydrogen peroxide were significantly different in the crystal and no crystal group at different time intervals. Enzyme Linked Immuno Sorbent Assay of important genes such as Kidney Injury Molecule, Monocyte chemoattractant protein and Osteopontin also showed significant difference between crystals and no crystals group at different time periods. CONCLUSIONS Our study provides a unique introspection into oxalate induced metabolic pathways and cellular processes going on in the cortex and medulla of rat's kidneys with and without crystals. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e854-e855 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Sunil Joshi Gainesville, FL More articles by this author Wei Wang Gainesville, FL More articles by this author Ammon Peck Gainesville, FL More articles by this author Saeed Khan Gainesville, FL More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...