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Sub-lethal concentrations of CdCl2 disrupt cell migration and cytoskeletal proteins in cultured mouse TM4 Sertoli
Biola F Egbowon1, Wayne Harris1, Gordon Arnott1
1School of Science and Technology, Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS, United Kingdom.
Abstract:
The aims of this study were to examine the effects of CdCl2 on the viability, migration and cytoskeleton of cultured mouse TM4 Sertoli cells. Time- and concentration-dependent changes were exhibited by the cells but 1 μM CdCl2 was sub-cytotoxic at all time-points. Exposure to 1 and 12 μM CdCl2 for 4 h resulted in disruption of the leading edge, as determined by chemical staining. Cell migration was inhibited by both 1 and 12 μM CdCl2 in a scratch assay monitored by live cell imaging, although exposure to the higher concentration was associated with cell death. Western blotting and immunofluorescence staining indicated that CdCl2 caused a concentration dependent reduction in actin and tubulin levels. Exposure to Cd(2+) also resulted in significant changes in the levels and/or phosphorylation status of the microtubule and microfilament destabilising proteins cofilin and stathmin, suggesting disruption of cytoskeletal dynamics. Given that 1-12 μM Cd(2+) is attainable in vivo, our findings are consistent with the possibility that Cd(2+) induced impairment of testicular development and reproductive health may involve a combination of reduced Sertoli cell migration and impaired Sertoli cell viability depending on the timing, level and duration of exposure.
Insights
Cadmium chloride (CdCl2) exposure impacts mouse Sertoli cell viability and migration, disrupting the cytoskeleton. These findings suggest a potential mechanism for cadmium-induced reproductive health issues.
Area of Science:
- Reproductive Toxicology
- Cell Biology
- Environmental Health
Background:
- Sertoli cells are crucial for testicular development and spermatogenesis.
- Cadmium (Cd2+) is a toxic environmental pollutant linked to male reproductive dysfunction.
- Understanding the cellular mechanisms of cadmium toxicity is essential for reproductive health.
Purpose of the Study:
- To investigate the effects of cadmium chloride (CdCl2) on cultured mouse TM4 Sertoli cell viability, migration, and cytoskeleton.
- To determine the concentration- and time-dependent responses of Sertoli cells to CdCl2 exposure.
- To elucidate the molecular mechanisms underlying CdCl2-induced Sertoli cell impairment.
Main Methods:
- Cell viability assays
- Scratch migration assays with live cell imaging
- Western blotting for cytoskeletal proteins (actin, tubulin)
- Immunofluorescence staining for cytoskeletal organization
- Analysis of cofilin and stathmin protein levels and phosphorylation
Main Results:
- CdCl2 exposure caused time- and concentration-dependent effects on Sertoli cells.
- 1 μM CdCl2 was sub-cytotoxic, while higher concentrations (1-12 μM) disrupted cell leading edges and inhibited migration.
- Higher CdCl2 concentrations led to cell death and reduced actin and tubulin levels.
- CdCl2 altered cofilin and stathmin levels/phosphorylation, indicating cytoskeletal dynamics disruption.
- Observed effects were concentration-dependent, with 12 μM CdCl2 causing significant cell death.
Conclusions:
- CdCl2 impairs Sertoli cell viability and migration, potentially through cytoskeletal disruption.
- Cadmium-induced reproductive health issues may stem from reduced Sertoli cell migration and viability.
- Findings are relevant as Cd2+ levels observed in vitro are attainable in vivo.
