Free Radicals in Precision Cut Bovine Testicle Slices: Effects of 1,3,5 Trinitrobenzene Measured by Electron Paramagnetic Resonance.

Abstract : Precision cut tissue slices were prepared from bovine testes and incubated at 37 degrees C in Wymouth's media supplemented with the spin trap N-tert-butyl-a-nitrone or dimethyl sulfoxide. Slices were exposed for 0 to ih with 0-1000 micronmeter 1,3,5 trinitrobenzene (TNB). Radicals were detected in testis after glutathione depletion with diethylmaleate in control, TNB-treated and superoxide-treated slices. The hyperfine coupling constant of the TNB-induced PBN-radical adducts was aN = 1.629 mT and =0.333 mT. Analysis of variance showed more radicals were trapped in the TNB-treated slices 100 micronmeter-1000 micronmeter when compared to controls (P <0.0001). First order regression analysis of the dose response curve was y=O.04x+23(r2=O.66, n=30). This data shows the in vitro slice technique can be used to measure radicals induced by TNB in bovine testicles.