Use of Nucleic Acid-Protein Cytophotometry and Cytointerferometry in Characterizing Cellular Mode of Trichothecene Toxicant Action

摘要

Cytophotometric analyses were made of nucleic acid-protein responses in known and putative target tissues of adult male Sprague-Dawley rats injected intraperitoneally with T-2 toxin, a small molecular weight trichothecene mycotoxin at dosages of 0.75, 1.0, 1.5 and 6.0 mg/kg (1 LD 50 = 0.9 mg/kg). Animals were decapitated during acute (2-8 h) or delayed (7-28 d) intervals after exposure. Tissues removed and processed for subsequent microchemical and cytomorphological analyses included the heart, cecum, liver, kidney, lungs, spleen, thymus, adrenals and mesenteric spreads. Cellular RNA and protein levels were assayed using azure B-RNA and Coomassie-protein cytophotometry; DNA and chromatin alterations were quantified using Feulgen-DNA cytophotometry and ocular filar micrometry. Correlative biochemical data were obtained on changes in circulating histamine and corticosterone levels. In vitro studies were also conducted of nucleic acid-protection responses of cultured endothelial cells subjected to T-2 toxin using pyronin Y-fluorescence, flow cytofluorometry, azure B-RNA cytophotometry and cytointerferometric assay of protein.