Evaluation of DNA Damage in Different Stages of Mouse Spermatogenesis after Testicular X Irradiation

Cordelli, Eugenia; Fresegna, Anna Maria; Leter, Giorgio; Eleuteri, Patrizia; Spanò, Marcello; Villani, Paola

doi:10.1667/rr3053

Cited by 63 publications

(39 citation statements)

References 62 publications

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“…We previously showed that stem cells were not involved in the origin of delayed radiation effects in male germ cells [Cordelli et al, 2003]. In this study, the window of sensitivity for the induction of delayed effects in testicular cells and cauda spermatozoa points to differentiating spermatogonia as the main progenitors of spermatids with delayed damage, although we cannot exclude that preleptotene, zygotene, and early pachytene spermatocytes may have contributed (Fig.…”

Section: Discussionmentioning

confidence: 57%

“…This phenomenon has also been observed in male germ cells [de Boer et al, 2010]. Spermatogonial irradiation causes delayed genomic instability in descendant sperm [Sailer et al, 1995;Haines et al, 2001Haines et al, , 2002Cordelli et al, 2003] as well as in the progeny of irradiated male parents in rodents [Baulch and Raabe, 2005;Barber and Dubrova, 2006;Barber et al, 2009] and humans [Aghajanyan et al, 2011]. Remodeling of chromatin domains and nuclear matrix during spermiogenesis and in the zygote has been suggested to play a role in these phenomena [de Boer et al, 2010], but how and when they operate is still unclear.…”

Section: Introductionmentioning

confidence: 94%

“…Testis monocellular suspensions were obtained as described by Cordelli et al [2003]. Fat and connective tissue were removed and testes were minced and treated for 10 min with 1 mL 0.1% pepsin-HCl solution (pepsin, 1.5 mAnson units/mg; Serva, Heidelberg, Germany) at room temperature under continuous magnetic stirring.…”

Section: Flow Cytometric Dna Content Analysis Of Testis Cellsmentioning

confidence: 99%

“…Mechanistic understanding of non-targeted effects on the germ line is necessary to determine whether it is relevant for genetic risk assessment of radiation exposure. Mouse spermatozoa derived from irradiated premeiotic germ cells carry DNA strand breaks [Sailer et al, 1995;Haines et al, 2001Haines et al, , 2002Cordelli et al, 2003]. The persistence of these strand breaks is incompatible with the multiple mitotic cycles and checkpoints, meiosis and postmeiotic differentiation processes that take place in cells from the time of exposure to the generation of spermatozoa.…”

Section: Introductionmentioning

confidence: 99%

“…This suggests that DNA breaks in terminally differentiated cells may be the result of non-targeted radiation effects whose mechanism remains to be elucidated. We have hypothesized that DNA strand breaks present in mature spermatozoa derive from an active process of DNA fragmentation occurring during meiosis or subsequent spermiogenesis [Cordelli et al, 2003]. The aim of this study was to determine the stage of spermatogenesis at which DNA fragmentation occurs and investigate the pathways involved in this process.…”

Section: Introductionmentioning

confidence: 99%

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Direct and delayed X‐ray‐induced DNA damage in male mouse germ cells

Cordelli¹,

Eleuteri²,

Grollino³

et al. 2012

Environ and Mol Mutagen

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Sperm DNA integrity is essential for the accurate transmission of paternal genetic information. Various stages of spermatogenesis are characterized by large differences in radiosensitivity. Differentiating spermatogonia are susceptible to radiationinduced cell killing, but some of them can repair DNA damage and progress through differentiation. In this study, we applied the neutral comet assay, immunodetection of phosphorylated H2AX (g-H2AX) and the Sperm Chromatin Structure Assay (SCSA) to detect DNA strand breaks in testicular cells and spermatozoa at different times following in vivo X-ray irradiation. Radiation produced DNA strand breaks in testicular cells that were repaired within the first few hours after exposure. Spermatozoa were resistant to the induction of DNA damage, but non-targeted DNA lesions were detected in spermatozoa derived from surviving irradiated spermatogonia. These lesions formed while round spermatids started to elongate within the testicular seminiferous tubules. The transcription of pro-apoptotic genes at this time was also enhanced, suggesting that an apoptotic-like process was involved in DNA break production. Our results suggest that proliferating spermatogonia retain a memory of the radiation insult that is recognized at a later developmental stage and activates a process leading to DNA fragmentation. Environ. Mol. Mutagen. 53:429-439, 2012. V V C 2012 Wiley Periodicals, Inc.

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Section: Discussionmentioning

confidence: 57%

Section: Introductionmentioning

confidence: 94%

Section: Flow Cytometric Dna Content Analysis Of Testis Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Direct and delayed X‐ray‐induced DNA damage in male mouse germ cells

Cordelli¹,

Eleuteri²,

Grollino³

et al. 2012

Environ and Mol Mutagen

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The impact of testicular carcinoma and its treatment on sperm DNA integrity

Ståhl

Eberhard

Jepson

et al. 2004

Cancer

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BACKGROUNDIn patients with testicular germ cell carcinoma (TGCC), spermatogenesis and fertility are impaired. Intracytoplasmic sperm injection has improved their possibility of fatherhood, but might also impose a risk of transmitting DNA defects to the offspring. The aim of the current study was to evaluate the impact of chemotherapy and irradiation on sperm DNA integrity.METHODSThe study included 74 patients with TGCC. Semen samples were collected before and at specific time points after patients received therapy. Sperm DNA integrity was assessed by the sperm chromatin structure assay. Controls comprised 278 military conscripts.RESULTSThere was no significant difference in the fraction of sperm with fragmented DNA (DNA fragmentation index [DFI]) between controls and patients with TGCC before postoperative cancer treatment (11% vs. 13%). Men treated with adjuvant radiotherapy had a transiently (up to 2 years) higher DFI than nontreated patients (18% vs. 13%; P = 0.03). Patients who received 1–2 cycles of adjuvant chemotherapy had a significantly lower DFI 6 months after treatment than after 1–2 years (9.1% vs. 13%; P = 0.004). Higher doses of chemotherapy among patients resulted in a significantly lower DFI compared with controls (7.3% vs. 11%; P = 0.028), which persisted throughout the 5 years of follow‐up.CONCLUSIONSPostorchiectomy, the DFI in sperm samples from patients with testicular carcinoma was at the level of controls. Radiotherapy caused a transient increase in the proportion of DFI, whereas this value decreased after chemotherapy. The biologic implications of such changes in sperm DNA after cancer therapy need to be elucidated. Cancer 2004. © 2004 American Cancer Society.

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Spermatocyte responses in vitro to induced DNA damage

Matulis

Handel

2006

Molecular Reproduction Devel

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Spermatocytes normally sustain many meiotically induced double-strand DNA breaks (DSBs) early in meiotic prophase; in autosomal chromatin, these are repaired by initiation of meiotic homologous-recombination processes. Little is known about how spermatocytes respond to environmentally induced DNA damage after recombination-related DSBs have been repaired. The experiments described here tested the hypothesis that, even though actively completing meiotic recombination, pachytene spermatocytes cultured in the absence of testicular somatic cells initiate appropriate chromatin remodeling and cell-cycle responses to environmentally induced DNA damage. Two DNA-damaging agents were employed for in vitro treatment of pachytene spermatocytes: gamma-irradiation and etoposide, a topoisomerase II (TOP2) inhibitor that results in persistent unligated DSBs. Chromatin modifications associated with DSBs were monitored after exposure by labeling surface-spread chromatin with antibodies against RAD51 (which recognizes DSBs) and the phosphorylated variant of histone H2AFX (herein designated by its commonly used symbol, H2AX), gammaH2AX (which modifies chromatin associated with DSBs). Both gammaH2AX and RAD51 were rapidly recruited to irradiation- or etoposide-damaged chromatin. These chromatin modifications imply that spermatocytes recruit active DNA damage responses, even after recombination is substantially completed. Furthermore, irradiation-induced DNA damage inhibited okadaic acid-induced progression of spermatocytes from meiotic prophase to metaphase I (MI), implying efficacy of DNA damage checkpoint mechanisms. Apoptotic responses of spermatocytes with DNA damage differed, with an increase in frequency of early apoptotic spermatocytes after etoposide treatment, but not following irradiation. Taken together, these results demonstrate modification of pachytene spermatocyte chromatin and inhibition of meiotic progress after DNA damage by mechanisms that may ensure gametic genetic integrity.

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Evaluation of DNA Damage in Different Stages of Mouse Spermatogenesis after Testicular X Irradiation

Cited by 63 publications

References 62 publications

Direct and delayed X‐ray‐induced DNA damage in male mouse germ cells

Direct and delayed X‐ray‐induced DNA damage in male mouse germ cells

The impact of testicular carcinoma and its treatment on sperm DNA integrity

Spermatocyte responses in vitro to induced DNA damage

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