Flow cytometry (FCM) was performed to monitor the cellular effects of extremely-low-frequency magnetic field on mouse spermatogenesis. Groups of five male hybrid F1 mice aged 8-10 weeks were exposed to 50 Hz magnetic field. The strength of the magnetic field was 1.7 mT. Exposure times of 2 and 4 h were chosen. FCM measurements were performed 7, 14, 21, 28, 35, and 42 days after treatment. For each experimental point, a sham-treated group was used as a control. The possible effects were studied by analyzing the DNA content distribution of the different cell types involved in spermatogenesis and using the elongated spermatids as the reference population. The relative frequencies of the various testicular cell types were calculated using specific software. In groups exposed for 2 h, no effects were observed. In groups exposed for 4 h, a statistically significant (P < 0.001) decrease in elongated spermatids was observed at 28 days after treatment. This change suggests a possible cytotoxic and/or cytostatic effect on differentiating spermatogonia. However, further studies are being carried out to investigate the effects of longer exposure times.
The effects of heat on mouse spermatogenesis have been determined using both testis weight and flow cytometrically determined DNA content distribution as experimental end-points. Temperatures of 38-42 degrees C and exposure times of 20-60 min have been tested. The results concerning the testis weight substantially confirm those reported by other authors (Hand et al. 1979). The measurement of DNA content distributions shows a relatively higher depletion, 14 days after treatment, of the cytometric compartment containing elongated spermatids in respect to that containing round spermatids. The analysis of the cytotoxic effects, monitored 14 vs. 28 days after treatment, as a function of the exposure time at a given temperature, or of the temperature for a fixed exposure time, indicates that, in the course of spermatogenesis, late spermatocytes are more sensitive to heat than differentiated spermatogonia. Following the approach based on flow cytometry, the effect of exposures as low as 20 min at 38 degrees C can be appreciated.
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