2-DIMENSIONAL ELECTROPHORESIS OF PRECISION-CUT TESTIS SLICES - TOXICOLOGIC APPLICATION

Advances in tissue slice technology and a recent novel application of this technique to reproductive toxicology using bovine testis have dem onstrated the remarkable utility of this approach. The objective of th e present study was to combine this in vitro toxicity test system with large-scale two-dimensional polyacrylamide gel electrophoresis (2-DE) to detect and study alterations in testicular-slice protein patterns as molecular correlates of 1,3,5-trinitrobenzene (TNB) and 1,3-dinitro benzene (DNB) toxicity. Previous studies have shown that testicular sl ices remain viable for > 24 h and, as measured by protein synthesis in hibition, TNB causes dose-related injury. Tissue-slices were prepared from bovine testicles incubated for 2, 4 or 6 h and exposed to either 100 mu M, 500 mu M Or 1 mM DNB or TNB in the incubation medium. Slices were collected, solubilized, and separated by large scale 2-DE, Resul ting protein patterns were then examined by image analysis, which reve aled coefficients of variation in protein spot abundance comparable to patterns from fresh rodent tissue samples. Furthermore, specific prot ein alterations indicated dose-related inductions and declines in prot ein abundance, some progressive over time. The results of this investi gation demonstrate the potential toxicologic utility of combining in v itro tissue-slice technology with high-resolution: 2-DE protein mappin g. The consolidation of these methods offers a novel approach for toxi city screening and testing, reduces experimental cost, and reduces the use of laboratory animals.