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Flow cytometric assessment of trophosphamide toxicity on mouse spermatogenesis
M Spanò1, C Bartoleschi, E Cordelli
1Division of Environmental Toxicology, ENEA Casaccia, Rome, Italy.
Abstract:
The effects of trophosphamide on mouse reproductive cells have been investigated by flow cytometric analysis of testicular cell populations and alterations of sperm chromatin structure. Mice were treated with single intraperitoneal injections of TP, the doses ranging between 50 and 150 mg/kg, and were killed after 7, 14, 21, 28, 35, or 49 days. Dose-dependent reductions of tetraploid cells, round spermatids, and elongated spermatids were detected at 7, 21, and 28 days, respectively, reflecting cytotoxic damage to the differentiating spermatogonia compartment. The dose necessary to reduce the number of differentiating spermatogonia to half the control value was approximately 70 mg/kg. Stem cells were not affected by this treatment, and the normal spermatogenic process was restored after 7 weeks. In addition, cauda epididymal sperm were analyzed by the sperm chromatin structure assay, a flow cytometric measurement of the susceptibility of the sperm nuclear DNA to in situ acid denaturation; a statistically significant increase of sperm with altered chromatin structure was detected after a TP treatment of 150 mg/kg. Together with previous findings published in the literature, where the same doses induced heritable genetic damage, this study demonstrates a marked adverse cytotoxic effect of TP on the male reproductive integrity. All this information should be taken into consideration when TP is used in chemotherapeutic regimens.
Insights
Trophosphamide (TP) causes significant cytotoxic damage to mouse reproductive cells, reducing spermatogonia and altering sperm DNA. This highlights TP
Area of Science:
- Reproductive toxicology
- Chemotherapy drug effects
- Spermatogenesis research
Background:
- Trophosphamide (TP) is a chemotherapeutic agent.
- Understanding its impact on male reproductive health is crucial.
- Previous studies indicated potential for heritable genetic damage.
Purpose of the Study:
- To investigate the cytotoxic effects of TP on mouse testicular cell populations.
- To assess alterations in sperm chromatin structure following TP exposure.
- To evaluate the impact of TP on male reproductive integrity.
Main Methods:
- Flow cytometric analysis of testicular cell populations.
- Sperm chromatin structure assay (SCSA) on cauda epididymal sperm.
- Mice treated with single intraperitoneal injections of TP (50-150 mg/kg) and analyzed at various time points (7-49 days).
Main Results:
- Dose-dependent reductions in tetraploid cells, round spermatids, and elongated spermatids were observed.
- Cytotoxic damage was evident in the differentiating spermatogonia compartment.
- A significant increase in sperm with altered chromatin structure was detected at the highest TP dose (150 mg/kg).
- Stem cells remained unaffected, and spermatogenesis recovered within 7 weeks.
Conclusions:
- Trophosphamide exhibits a marked adverse cytotoxic effect on male reproductive integrity.
- TP treatment leads to significant damage to spermatogonia and sperm chromatin.
- Findings necessitate careful consideration of TP's reproductive toxicity in chemotherapeutic regimens.