High-content Analysis Provides Mechanistic Insights into the Testicular Toxicity of Bisphenol A and Selected Analogues in Mouse Spermatogonial Cells

Bisphenol A (BPA), an endocrine-disrupting compound, was found to be a testicular toxicant in animal models. Bisphenol S (BPS), bisphenol AF (BPAF), and tetrabromobisphenol A (TBBPA) were recently introduced to the market as alternatives to BPA. However, toxicological data of these compounds in the male reproductive system are still limited so far. The present study developed and validated an automated multi-parametric high-content analysis (HCA) using the C18-4 spermatogonial cell line as a model. We applied these validated HCA, including nuclear morphology, DNA content, cell cycle progression, DNA synthesis, cytoskeleton integrity, and DNA damage responses, to characterize and compare the testicular toxicities of BPA and three selected commercial available BPA analogues, BPS, BPAF and TBBPA. HCA revealed BPAF and TBBPA exhibited higher spermatogonial toxicities as compare with BPA and BPS, including dose- and time-dependent alterations in nuclear morphology, cell cycle, DNA damage responses, and perturbation of the cytoskeleton. Our results demonstrated that this specific culture model together with HCA can be utilized for quantitative screening and discriminating of chemical-specific testicular toxicity in spermatogonial cells. It also provides a fast and cost-effective approach for the identification of environmental chemicals that could have detrimental effects on reproduction.

Toxicological sciences : an official journal of the Society of Toxicology. 2016 Sep 14 [Epub ahead of print]

Shenxuan Liang, Lei Yin, Kevin Shengyang Yu, Marie-Claude Hofmann, Xiaozhong Yu

1. Department of Environmental Health Science, College of Public Health, University of Georgia, Athens, Georgia 30602., 1. Department of Environmental Health Science, College of Public Health, University of Georgia, Athens, Georgia 30602 2. ReproTox Biotech LLC, 111 Riverbend Drive, Athens, Georgia 30602., 3. Department of Endocrine Neoplasia & Hormonal Disorders, MD Anderson Cancer Center, 1515 Holcombe Blvd, Unit # 1105 Houston, TX 77030., 1. Department of Environmental Health Science, College of Public Health, University of Georgia, Athens, Georgia 30602 .