Effects of heat on mouse spermatogenesis monitored by flow cytometry

Vita, R. De; Calugi, A; Chiarantano, C.; Forte, D.; Mauro, F.; Uccelli, Raffaella

doi:10.3109/02656739009140950

Cited by 20 publications

(10 citation statements)

References 26 publications

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“…Spermatozoa ejaculated 21 days after heating would have been pachytene and diplotene spermatocytes at the time of heating [42,43] and these cells have been previously reported to be seriously damaged by heat exposure [45,56,57] and this study. Apparently this damage can extend to an effect on the development of preimplantation embryos.…”

Section: Discussionmentioning

confidence: 74%

Effects of paternal heat stress on the in vivo development of preimplantation embryos in the mouse

Zhu

Setchell

2004

Reprod. Nutr. Dev.

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-The objective of this study was to examine the effect of paternal heat stress on the in vivo development of preimplantation embryos in the mouse. Synchronised B6CBF1 female mice were mated either to a control male mouse or to one that had been exposed at 7, 21 or 35 days previously, for 24 h to an ambient temperature of 36 ± 0.3 °C and 66 ± 5.6% relative humidity. Embryos were collected from the oviducts of mice at 14-16 h, 34-39 h or 61-65 h after mating or from the uterus at 85-90 h after mating and their developmental status was evaluated morphologically. The number of cells within blastocysts was also determined using bisbenzimide-propidium iodide staining. Paternal heat stress 7 days before mating reduced the proportion of embryos developing from 4-cell (4-C) to morulae (M), hatched blastocysts, total blastocysts and the number of inner cell mass (ICM) and trophectoderm (TE) cells in the blastocyst. Paternal heat stress 21 days prior to mating reduced the proportion of 2-C and 4-C to M embryos with no embryos developing to blastocysts. There were also increases in the number of 1-C and abnormal embryos recorded at this time. Paternal heat stress 35 days before mating decreased the proportion of 2-C embryos, expanded blastocysts and ICM and TE cells in the blastocyst. These results support previous work demonstrating that both the sperm in the epididymis and germ cells in the testis are susceptible to damage by environmental heat stress, with spermatocytes being the most vulnerable. This study also demonstrates that subtle effects on the male such as a short exposure to elevated environmental temperatures can translate to quite profound paternal impacts on early embryo development.paternal heat stress / in vivo development / mouse preimplantation embryos

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Section: Discussionmentioning

confidence: 74%

Effects of paternal heat stress on the in vivo development of preimplantation embryos in the mouse

Zhu

Setchell

2004

Reprod. Nutr. Dev.

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“…Previous studies have identified pachytene spermatocytes and early spermatids as being susceptible to heat stress (Collins & Lacy 1969, De Vita et al 1990) and this is not surprising as during meiotic prophase substantial rearrangements of DNA occurs within the nuclei of the leptotene, zygotene and pachytene spermatocytes. Collins & Lacy (1969) identified two critical periods in spermatogenesis (leptotene-pachytene and maturation division) through which cells were unable to progress following heating.…”

Section: Discussionmentioning

confidence: 99%

“…In an extensive study by De Vita et al (1990), it was shown that exposure to mild heat stress (38 8C) for a short time (20 min) caused cytotoxic effects in some germ cell types and that these effects were more pronounced in spermatocytes than in differentiated spermatogonia. Following heat stress, the epididymis was reported to lose its ability to store and maintain viable spermatozoa, resulting in the gradual and progressive accumulation of dead, decapitated and immotile spermatozoa (Bedford 1991(Bedford , 1994; the region most affected is reported to be the cauda.…”

Section: Introductionmentioning

confidence: 99%

Impact of a mild scrotal heat stress on DNA integrity in murine spermatozoa

Banks¹,

King²,

Irvine³

et al. 2005

Reproduction

218

125

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An increase in scrotal temperature can lead to the production of poor quality spermatozoa and infertility. In the present study we have used mice to examine the impact of mild, scrotal heat stress (42 8C for 30 min) on numbers of spermatozoa as well as on the integrity of their DNA. Spermatozoa recovered from the epididymides hours (1 to 24) or days (7 to 32) after treatment were analysed using COMET and sperm chromatin structure (SCSA) assays. The treatment induced a stress response in both the testis and the epididymis that was associated with reduced expression of the cold inducible RNA binding protein (Cirp) and an increase in germ cell apoptosis (Apotag positive cells). Although spermatozoa present in the epididymis at the time of heating contained correctly packaged DNA, its integrity was compromised by heat stress. In addition, although some germ cells, which were present within the testis at the time of heat stress, were removed by apoptosis, many germ cells completed their development and were recovered as motile spermatozoa with damaged DNA. In conclusion, these data demonstrate that scrotal heat stress can compromise the DNA integrity of spermatozoa and this may have clinical implications for patients undergoing IVF and intra-cytoplasmic sperm injection (ICSI).

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“…Therefore, we deemed it interesting to use this experimental system and this analytical approach to investigate possible cytotoxic and/or cytostatic effects induced by ELF magnetic field at the frequency of 50 Hz. A wide range of noxious agents can damage any step in spermatogenesis, and flow cytometry has been used to evaluate variations in the relative frequency of the different cell populations [Meistrich et al, 1978;HackerKlom et al, 1981HackerKlom et al, , 1984HackerKlom et al, , 1985Otto et al, 1981Otto et al, , 1984Spanb et al, 1987;De Vita et al, 1990 The elongated spermatids were used in this study as a reference population to evaluate effects on specific cells in the spermatogenic pathway. The flow cytometric frequency of this population is highly reproducible and stable, without cyclic variations, as confirmed by the previously cited studies.…”

Section: Discussionmentioning

confidence: 99%

Effects of 50 Hz magnetic fields on mouse spermatogenesis monitored by flow cytometric analysis

Vita

Cavallo

Raganella

et al. 1995

Bioelectromagnetics

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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.

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Effects of heat on mouse spermatogenesis monitored by flow cytometry

Cited by 20 publications

References 26 publications

Effects of paternal heat stress on the in vivo development of preimplantation embryos in the mouse

Effects of paternal heat stress on the in vivo development of preimplantation embryos in the mouse

Impact of a mild scrotal heat stress on DNA integrity in murine spermatozoa

Effects of 50 Hz magnetic fields on mouse spermatogenesis monitored by flow cytometric analysis

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