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Two‐dimensional electrophoresis of precision‐cut testis slices: Toxicologic application
Author(s) -
Witzmann Frank A.,
Fultz Carla D.,
Wyman John F.
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150180350
Subject(s) - toxicity , in vitro , gel electrophoresis , incubation , polyacrylamide gel electrophoresis , biology , chromatography , chemistry , microbiology and biotechnology , toxicology , biochemistry , enzyme , organic chemistry
Advances in tissue slice technology and a recent novel application of this technique to reproductive toxicology using bovine testis have demonstrated the remarkable utility of this approach. The objective of the 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‐dinitrobenzene (DNB) toxicity. Previous studies have shown that testicular slices remain viable for > 24 h and, as measured by protein synthesis inhibition, TNB causes dose‐related injury. Tissue‐slices were prepared from bovine testicles incubated for 2, 4 or 6 h and exposed to either 100 μ M , 500 μ M or 1 m M DNB or TNB in the incubation medium. Slices were collected, solubilized, and separated by large scale 2‐DE. Resulting protein patterns were then examined by image analysis, which revealed coefficients of variation in protein spot abundance comparable to patterns from fresh rodent tissue samples. Furthermore, specific protein alterations indicated dose‐related inductions and declines in protein abundance, some progressive over time. The results of this investigation demonstrate the potential toxicologic utility of combining in vitro tissue‐slice technology with high‐resolution 2‐DE protein mapping. The consolidation of these methods offers a novel approach for toxicity screening and testing, reduces experimental cost, and reduces the use of laboratory animals.