ABSTRACT
T-2 toxin is the most toxic among mycotoxins and poses a potential health hazard for both humans and animals. At high doses, T-2 toxin can cause shock-like syndrome that can result in death. We evaluated the effect of time course and route of exposure on hepatic oxidative damage in mice and it is only such study so far to compare the effects of dermal and subcutaneous exposure of T-2 toxin. Mice were exposed to 1 LD50 of T-2 toxin either by percutaneous (5.94 mg/kg body weight) or subcutaneous (1.54 mg/kg body weight) route and sacrificed at 0, 1, 3, and 7 days postexposure. Analysis of a number of serum biochemical variables, antioxidant enzymes activity, gene and protein expression by immunoblot assay showed time and route dependent effects of T-2 induced hepatic oxidative damage. Time dependent increase in protein carbonyl content and protein oxidation was seen in serum and liver. Results of our study may provide possible mechanism for developing medical countermeasures against T-2 toxin.
Subject(s)
Antioxidants/metabolism , Liver/drug effects , Oxidative Stress/drug effects , T-2 Toxin/toxicity , Administration, Cutaneous , Animals , Biomarkers/blood , Catalase/genetics , Catalase/metabolism , Female , Gene Expression/drug effects , Glutathione Peroxidase/genetics , Glutathione Peroxidase/metabolism , Injections, Subcutaneous , Liver/enzymology , Liver/metabolism , Mice , Protein Carbonylation , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolismABSTRACT
T-2 toxin belongs to group of mycotoxins and is found as a natural contaminant in cereals, feed and vegetables. In the present study we evaluated acute toxicity of dermal and subcutaneous exposure of T-2 toxin on brain oxidative stress in mice. Mice were exposed to 1 LD50 of T-2 toxin either by dermal (5.94 mg/kg) or subcutaneous (1.54 mg/kg body weight) route and sacrificed at 1, 3 and 7 days post-exposure. T-2 toxin treated animals showed time dependent increase in reactive oxygen species generation, glutathione depletion, lipid peroxidation and protein carbonyl content in brain in both the routes of exposure. Gene expression profile of antioxidant enzymes showed significant increase in superoxide dismutase and catalase in percutaneous route and glutathione reductase and glutathione peroxidase in subcutaneous route. Immunoblot analysis of antioxidant enzymes correlated with gene expression profile. T-2 toxin exposure resulted in down regulation of transcription factor Nrf2 and its downstream target genes of phase II detoxifying enzymes NQO1, Gclc, Gclm and hemeoxygenase-1. Results of our study show that percutaneously and subcutaneously applied T-2 toxin can cause brain oxidative damage possibly after crossing blood-brain barrier by altering its permeability.
Subject(s)
Brain Chemistry/drug effects , Oxidative Stress/drug effects , T-2 Toxin/toxicity , Administration, Topical , Animals , Antioxidants/metabolism , Blotting, Western , Body Weight/drug effects , Female , Gene Expression/drug effects , Glutathione/metabolism , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Injections, Subcutaneous , Lipid Peroxidation/drug effects , Mice , Nerve Tissue Proteins/drug effects , Oxidation-Reduction , Protein Carbonylation/drug effects , RNA/biosynthesis , RNA/genetics , Reverse Transcriptase Polymerase Chain Reaction , T-2 Toxin/administration & dosageABSTRACT
The zona pellucida (ZP) glycoproteins play an important role in oocyte development and gamete biology. To analyze their expression in follicles during various developmental stages, murine monoclonal antibodies (MAbs) were generated against the baculovirus-expressed recombinant human ZP2, ZP3 and ZP4. A panel of MAbs specific for the respective zona protein in ELISA and Western blot, and devoid of cross-reaction with other zona proteins was selected. Immunohistochemistry has shown that ZP2 MAb, MA-1620, did not react with oocytes in resting primordial follicles but showed reactivity with degenerating oocytes in primordial follicles undergoing atresia, and with oocytes in growing and antral follicles. Three MAbs against ZP3 did not react with oocytes in primordial follicles, but reacted only with oocytes in growing and antral follicles. Out of four MAbs against ZP4, three MAbs reacted with oocytes in primordial, growing and antral follicles. No reactivity of these MAbs with other ovarian cell types and other tissues studied (endometrium, uterine cervix, fallopian tubes and kidney) was detected except for a strong reactivity of ZP2 MA-1620 with epithelial cells of the uterine ectocervix or endometrium in some samples investigated. Altogether, these studies document generation of MAbs exhibiting high specificity for human zona proteins, which will be useful reagents to study their immunobiology.
