Proteomic analysis in body fluids and soft tissues in rats submitted to treatment with fluoride

Abstract

The recent development of proteomic techniques has allowed the analysis of the entire protein profile of the biological systems, contributing to the understanding of the normal physiology of the organism, as well as the mechanisms of diseases and the discovery of biomarkers for their early detection, identification of new therapies and drugs. This study will use proteomic techniques to analyze the differential protein expression in kidney and plasma of rats submitted to acute or chronic fluoride doses. In the first part of the study, three groups of five 75-day-old Wistar rats each will receive by gastric gavage the following single doses of fluoride: 0 (control), 50 and 100 mgF/Kg bw. In the second part of the study, 3 groups of 5 animals each will be received as weanlings and will drink, for 60 days, water containing 0 (control), 5 or 50 ppm F. After these treatments the animals will be euthanized and the kidneys (cortex and medulla separately) and serum will be collected. Fluoride in plasma and renal tissues will be analyzed with the electrode, after hexamethyldisiloxane-facilitated diffusion. Proteins from kidney, serum and urine will be profiled by two-dimensional gel electrophoresis. Gels from control and treated groups will be digitalized using the ImageScanner and analyzed with ImageMaster 2D Platinum software. Spots of interest will be excised for protein identification by mass spectrometry (MALTI-TOF). The effect of fluoride on the activity of protein phosphatases of renal cells and the phospho-tyrosin proteins will also be determined by Western blotting.