Two-dimensional electrophoresis of precision-cut testis slices: Toxicologic application

Frank Witzmann, Carla D. Fultz, John F. Wyman

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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 μM, 500 μM or 1 mM 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.

Original languageEnglish (US)
Pages (from-to)642-646
Number of pages5
JournalElectrophoresis
Volume18
Issue number3-4
DOIs
StatePublished - Mar 1997

Fingerprint

Electrophoresis
Testis
Proteins
Tissue
Toxicity
Technology
Reproductive Techniques
Toxicity Tests
Electrophoresis, Gel, Two-Dimensional
Laboratory Animals
Consolidation
Toxicology
Image analysis
Rodentia
Screening
Animals
Costs and Cost Analysis
Wounds and Injuries
Testing
sym-trinitrobenzene

Keywords

  • Dinitrobenzene
  • Protein mapping
  • Testis
  • Tissue slice
  • Toxicity test
  • Trinitrobenzene
  • Two-dimensional electrophoresis

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Two-dimensional electrophoresis of precision-cut testis slices : Toxicologic application. / Witzmann, Frank; Fultz, Carla D.; Wyman, John F.

In: Electrophoresis, Vol. 18, No. 3-4, 03.1997, p. 642-646.

Research output: Contribution to journalArticle

Witzmann, Frank ; Fultz, Carla D. ; Wyman, John F. / Two-dimensional electrophoresis of precision-cut testis slices : Toxicologic application. In: Electrophoresis. 1997 ; Vol. 18, No. 3-4. pp. 642-646.
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