Related Experiment Video
Updated: Mar 16, 2026
Functional Assessment of Kinesin-7 CENP-E in Spermatocytes Using In Vivo Inhibition, Immunofluorescence and Flow Cytometry
Published on: December 28, 2021
2,2',4,4'-Tetrabromodiphenyl ether injures cell viability and mitochondrial function of mouse spermatocytes by
Shaoping Huang1, Jing Wang2, Yiqiang Cui3
1Department of Human Anatomy and Neuroscience, Medical School, Southeast University, Nanjing 210009, Jiangsu, China; State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, Jiangsu, China.
Abstract:
Our object was to explore direct effects and mechanism of BDE47 on GC2 (immortalized mouse spermatocyte). GC2 were exposed to DMSO, 0.1, 1, 10, 100μM BDE47 for 48h. Cell viability was detected by trypan-blue exclusion; ultrastructure by electron-microscopy; cell cycle, mitochondrial membrane motential (MMP), reactive oxygen species (ROS) by flow-cytometry; ATP production by luminometer; Atp5b, Uqcrc1, Bcl-2 level by WB. To explore whether the decreased mitochondrial proteins play an important role in apoptosis, MMP and apoptosis were detected after Atp5b or Uqcrc1 knockdown in GC2. Results showed BDE47 reduced cell viability, caused condensation of nuclear and vacuolated mitochondria, decreased MMP and ATP, induced ROS, cell cycle arrest at S and G2/M phase, reduced Atp5b, Uqcrc1, Bcl-2 in GC2. Knockdown of Atp5b or Uqcrc1 decreased MMP, induced apoptosis in GC2. Results suggested that BDE47 reduced cell viability, injured mitochondria in spermatocytes probably by decreasing mitochondrial protein Atp5b and Uqcrc1.
Insights
Polybrominated diphenyl ether BDE47 exposure harms mouse spermatocyte viability and mitochondrial function. This damage is linked to reduced mitochondrial proteins, suggesting BDE47
Area of Science:
- Environmental Toxicology
- Reproductive Biology
- Cell Biology
Background:
- Polybrominated diphenyl ethers (PBDEs) are environmental contaminants.
- PBDEs, like BDE47, raise concerns regarding reproductive health.
- Spermatocytes are crucial for male fertility and susceptible to toxic insults.
Purpose of the Study:
- To investigate the direct toxic effects of BDE47 on mouse spermatocyte (GC2) viability.
- To elucidate the underlying mechanisms of BDE47-induced testicular toxicity.
- To determine the role of mitochondrial dysfunction and specific proteins in BDE47 toxicity.
Main Methods:
- GC2 cells were exposed to varying concentrations of BDE47.
- Cell viability, ultrastructure, cell cycle, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) were assessed.
- ATP production, and protein levels of Atp5b, Uqcrc1, and Bcl-2 were measured.
- Knockdown experiments for Atp5b and Uqcrc1 were performed to assess their role in apoptosis.
Main Results:
- BDE47 exposure reduced GC2 cell viability and induced nuclear condensation and mitochondrial vacuolation.
- BDE47 decreased MMP, ATP production, and increased ROS levels.
- Cell cycle arrest at S and G2/M phases was observed.
- BDE47 reduced levels of mitochondrial proteins Atp5b and Uqcrc1, and anti-apoptotic protein Bcl-2.
- Knockdown of Atp5b or Uqcrc1 diminished MMP and promoted apoptosis.
Conclusions:
- BDE47 exerts direct toxic effects on mouse spermatocytes, leading to reduced viability.
- Mitochondrial injury, evidenced by decreased MMP and ATP, and increased ROS, is a key mechanism.
- Reduced expression of mitochondrial proteins Atp5b and Uqcrc1 contributes to BDE47-induced apoptosis.
- BDE47 poses a risk to male reproductive health through testicular toxicity.
