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From the Departamento de Reproducción Animal y Conservación de Recursos Zoogenéticos, Instituto Nacional de Investigación y Tecnologia Agraria y Alimentaria, Carretera de la Coruña, Madrid, Spain.
| Correspondence to: Dr Alfonso Gutiérrez-Adán, Instituto Nacional de Investigación y Tecnologia Agraria y Alimentaria, Departamento de Reproducción Animal, Carretera de la Coruña km 5, 9, 28040 Madrid, Spain (e-mail: agutierr{at}inia.es). |
| Received for publication April 27, 2007; accepted for publication July 30, 2007. |
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Abstract |
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Key words: Sperm endonucleases, DNA integrity, ICSI, embryo quality
Recently the presence of endogenous nucleases that cleave spermatozoa DNA in mouse sperm has been reported; the nucleases can be activated by freeze-thawing of the spermatozoa in media without cryoprotectant (Sotolongo et al, 2005). The presence of these nucleases may produce unexpected sperm DNA fragmentation during the in vitro incubation period prior to in vitro fertilization (IVF) or ICSI if some of the spermatozoa have damaged membranes and, as a consequence, reduce the efficiency of the technology or induce long-term undesirable effects. In a recent report, it was demonstrated that in vitro incubation of swim-up-selected human spermatozoa in human tubal fluid medium without the addition of external factors leads to a progressive increase in the percentage of male gametes with fragmented DNA (Muratori et al, 2003). To explain spontaneous DNA fragmentation during in vitro sperm incubation, Maione et al (1997) postulated the involvement of sperm endogenous endonuclease activity. These authors reported that a particular nuclease activity is present in mature sperm. Subsequently, Ward and Ward (2004) hypothesized that the spontaneous DNA degradation within the sperm nucleus could be due to the enzymatic activity of endonucleases released from sperm with plasma membrane damage. Concomitantly, supporting evidence was reported of the existence of an endogenous nuclease in hamster, mouse, and human spermatozoa that cleaves DNA into loop-sized fragments (Sotolongo et al, 2005).
Our aims in this study were to evaluate if factors released from membrane-fragmented spermatozoa can act in the process of sperm DNA degradation and to examine the consequences on embryo development after ICSI with spermatozoa exposed to these factors. To achieve these goals, viability and DNA integrity of the sperm samples (levels of DNA fragmentation) were evaluated; ICSI was performed with sperm samples showing different levels of DNA fragmentation, and the subsequent development, implantation, and offspring obtained were analyzed.
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Materials and Methods |
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Animals
Hybrid B6D2F1 mice (Harlan Iberica SL, Barcelona, Spain) were used as
gamete donors. Females were 6 to 8 weeks old at the time of the experiments,
and males were at least 2 months old. CD1 females were used as surrogate
mothers for embryo transfer experiments after they were mated with
vasectomized CD1 males. Mice were fed ad libitum with a standard diet and
maintained in a temperature- and light-controlled room (23°C; 14 hours
light:10 hours dark). All animal experiments were approved by the
institutional review board of the Instituto Nacional de Investigación y
Tecnologia Agraria y Alimentaria according to the Guide for Care and Use of
Laboratory Animals as adopted by the Society for the Study of
Reproduction.
Preparation of Spermatozoa
To obtain the sperm cells, B6D2 10- to 12-week-old male mice were killed by
cervical dislocation. Cauda epididymides and vasa deferentia were placed in
500 µL of M2 medium, and adipose tissue and blood vessels were removed.
Then clean structures were placed in a new 1-mL drop of M2 medium covered with
mineral oil (Sigma) in which spermatozoa were collected. Concentrations were
determined with a Bürker hemocytometer. Four incubation conditions were
assayed: 1) spermatozoa were analyzed immediately after collection (no
incubation), 2) spermatozoa were incubated for 90 minutes in M2 medium, 3)
spermatozoa were incubated for 90 minutes in conditioned medium (CM; medium
that contained the different factors that spermatozoa release when their
membranes are damaged), and 4) spermatozoa were incubated for 90 minutes in CM
in the presence of 0.05 M EDTA, an ion chelator.
Preparation of CM— Spermatozoa collected from epididymides and vasa deferentia of B6D2 male mice were placed in 1-ml drops of M2 medium, sperm concentrations were determined with a Bürker hemocytometer, and samples were diluted if necessary to obtain final concentrations of 10 x 106 spermatozoa/mL. Sperm samples were placed into 1.5-mL tubes, frozen 3 times by plunging the tubes directly into liquid nitrogen in the absence of cryoprotectants, thawed at room temperature, incubated for 120 minutes at room temperature, and then centrifuged at 9300xg for 3 minutes to obtain a supernatant free of spermatozoa debris. This supernatant is what we called CM.
Incubation in CM— The incubation in CM was performed by adding 15 µL of fresh sperm into 300 µL of CM, followed by incubation for 90 minutes at room temperature.
Preparation of CM with 0.05 M EDTA— CM with 0.05 M EDTA was prepared as previously described, but in this case, M2 was modified by adding 25 mL of a 0.5 M solution of EDTA to 225 mL of M2 medium and reducing the concentration of NaCl to obtain a 285 to 295 mOsm medium (pH = 7.6). Incubation in CM with 0.05 M EDTA was performed as previously described for the incubation in CM.
In all sperm samples, we analyzed the percentage of viable spermatozoa and spermatozoa with DNA strand breaks according to the protocols described in the following sections.
Viability Assessment of Spermatozoa
Percentages of living and dead sperm cells were assessed using the staining
protocol of live/dead sperm viability kit (Molecular Probes, Eugene, Ore)
(Madrid-Bury et al, 2005)
Briefly, 0.8 µL of a 20 µM SYBR-14 working solution and 1.2 µL of a
2.4 mM propidium iodide (PI) working solution were added to 50 µL of the
sperm suspension (2–3 x 106 sperm cells/mL) and incubated at
37°C for 15 minutes. After that time, 20 µL of the sperm suspension was
loaded on a glass slide, covered with a coverslip, and observed immediately
under a fluorescent microscope equipped with the appropriate filters. The
SYBR-14 stained the nucleus of living sperm green, whereas dead or
membrane-damaged spermatozoa were stained red by PI, a conventional dead cell
nucleic acid stain (Garner and Johnson,
1995). At least 500 cells were counted for each treatment.
Determination of DNA Fragmentation in Mouse Sperm Cells
Gelled aliquots of 1% low melting point agarose in microfuge tubes were
placed in a water bath at 90°C to 100°C for 5 minutes to melt the
agarose and then transferred into a water bath at 37°C; after a 5-minute
incubation for temperature equilibration at 37°C, 30 µL of the sperm
sample was mixed with agarose (to obtain a 0.7% final agarose concentration).
Twenty microliters of the sperm-agarose mixture was then pipetted onto glass
slides precoated with 0.65% standard agarose, previously dried at 80°C,
and covered with a 22 x 22 mm coverslip. The slide was placed on a cold
plate in the refrigerator at 4°C for 5 minutes to allow the agarose to
produce a microgel with the sperm cells trapped within. The coverslips were
gently removed and the slide immediately immersed horizontally in 10 mL of a
solution that contained 4% β-mercaptoethanol (Sigma) and 0.05% Triton
X-100 (Sigma) in phosphate-buffered saline (PBS) to remove membranes and
reduce protamine–SS to –SH. After the slides were washed for 5
minutes with PBS, osmotic shock was induced by placing the slides in a 1 M
NaCl solution for 3 minutes. After the slides were washed in PBS for a further
5 minutes, they were placed in a 4% formaldehyde fixative solution for 30
minutes. Single- and double-strand DNA breaks were evaluated by the terminal
deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay using
the In Situ Cell Death Detection Kit (Roche, Mannheim, Germany) as described
by Pérez-Crespo et al
(2006). Slides were rinsed
twice in PBS and counterstained with 2 µg/mL 4,6 diamidoino-2-phenylindole
(DAPI) in Vectashield (Vector, Burlingame, Calif). Negative controls lacking
the TdT enzyme were run in each replicate. A minimum of 500 spermatozoa per
sample were scored under the 40x objective of the fluorescence
microscope (Nikon, Tokyo, Japan). The number of spermatozoa per field stained
with DAPI (blue) was first counted, and then the number of cells emitting
green fluorescence (TUNEL positive) was estimated; numbers were expressed as
percentages of the total cell count of the sample.
Oocyte Collection, ICSI, Embryo Culture, and Transfer
Meiosis II oocytes were collected 14 hours posthuman chorionic gonadotropin
(hCG) administration from 6- to 8-week-old female mice superovulated with 5 IU
of equine chorionic gonadotropin (Intervet, Boxmeer, The Netherlands),
followed 48 hours later by an equivalent dose of hCG (Lepori, Farma-Lepori,
Barcelona, Spain). Cumulus cells were dispersed by a 3- to 5-min incubation in
M2 medium containing 350 IU/mL hyaluronidase, and oocytes were washed and
maintained in potassium-modified simplex optimized medium (KSOM) at 37°C
in a 5% CO2 air atmosphere until use.
Fresh sperm samples (no incubation) and sperm samples incubated for 90 minutes in CM, collected from 8 B6D2 mice, were used to perform ICSI. All samples were centrifuged at 135xg for 3 minutes; in every sample, the supernatant was removed and the pellet was resuspended in 300 µL of M2 medium. One volume of mouse sperm sample was mixed with 5 of M2 containing 10% polyvinyl-pyrrolidine (PVP) to decrease stickiness. ICSI was performed as described (Moreira et al, 2004; Moreira et al, 2007). Briefly, ICSI was performed in M2 medium at room temperature. The ICSI dish contained a manipulation drop (M2 medium), a sperm drop (sperm solution in M2/10% PVP), and an M2/10% PVP needle-cleaning drop. Injections were performed with a PMM-150 FU piezo-impact unit (PrimeTech, Tokyo, Japan) and Eppendorf micromanipulators (Hamburg, Germany) using a blunt-ended mercury-containing pipette with 6 to 7 µm of inner diameter. The head of the fresh sperm cell was separated from the midpiece and tail by applying 1 or more piezoelectric pulses. Groups of 10 oocytes were injected with individual sperm heads. After 15 minutes of recovery at room temperature in M2 medium, surviving oocytes were washed 3 times in equilibrated KSOM and returned to mineral oil-covered KSOM drops and cultured at 37°C in a 5% CO2 air atmosphere.
Embryos were cultured in vitro until either the 2-cell stage (24 hours later) or the blastocyst stage (96 hours later) and then transferred into oviducts of pseudopregnant recipient females. Embryo transfer was performed as described previously (Gutiérrez-Adán et al, 2001). To be able to collect data on the total number of implantations, recipients were sacrificed on day 14, and both the number of fetuses and resorptions were recorded.
Statistical Analysis
Percentages of viable and TUNEL+ spermatozoa were compared by 1-way
repeated-measures analysis of variance (followed by multiple pairwise
comparisons using the Student-Newman-Kleus method). To normalize the
percentage data, arcsin square root quick transform was applied to the
percentages of TUNEL+ spermatozoa. The implantation rates and percentages of
fetuses were compared using the z-test. All the statistical analyses
described were performed using SigmaStat (Jandel Scientific, San Rafael,
Calif) software.
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Results |
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The results of DNA damage, detected by TUNEL, are illustrated in
Figure 1. When sperm cells were
incubated for 90 minutes in M2 medium, no significant increase in the
proportion of DFS was detected. However, when sperm cells were incubated for
90 minutes in CM, a significant increase in DFS was observed. When spermatozoa
were incubated in CM but in the presence of 0.05 M EDTA, no signs of increased
DNA damage were evident, suggesting that the substances responsible for
inducing DNA fragmentation in the spermatozoa are dependent on
ions.
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Preimplantation and Postimplantation Development of Mouse Embryos Produced by ICSI
To perform ICSI, we used either fresh spermatozoa or spermatozoa that had
been incubated for 90 minutes in CM. Embryos produced by ICSI were transferred
either at the 2-cell stage or as blastocysts after in vitro culture. Cleavage
rates, embryo development, and implantation rates of the embryos produced by
ICSI are shown in the Table. We did not find any significant differences in
the embryo developmental rate or morphology during the in vitro culture
period, indicating that both cleavage rate and blastocyst yield were not
affected by the incubation of spermatozoa with CM. However, the implantation
rate of embryos (either 2 cells or blastocysts) resulting from ICSI with
CM-pretreated sperm, was significantly reduced compared with those produced
with fresh sperm (z-test; P < .05) (Table). Moreover, the
significantly reduced implantation rate of blastocysts produced by sperm
pretreated with CM, which is lower than in any other group, suggests a
cumulative negative effect of ICSI with DFS and in vitro culture.
The recording of the number of day 14 fetuses represented the number of embryos that developed to term, and the number of resorptions gave us information about the number of postimplantation losses (embryos that were able to implant but could not develop to term). It is expected that natural selection ensures that most embryos with genetic damage will abort before growing to term. When we analyzed postimplantation development, we found that independently of sperm pretreatment, the in vitro culture significantly affected the percentage of fetuses obtained at day 14 of development (Figure 2). Moreover, we found a nonsignificant reduction in the percentage of fetuses at day 14 in the group of embryos resulting from ICSI with CM-pretreated sperm compared with fresh sperm when embryos were transferred at the 2-cell stage. Also, when embryos were cultured in vitro before embryo transfer, the number of resorptions was higher in the group of fresh sperm than in the group incubated with CM. This suggests that the in vitro culture reduced the implantation rate of embryos generated with fragmented DNA, whereas the negative effect of in vitro culture in the ICSI embryos fertilized with fresh sperm is observed later, during fetal development.
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Discussion |
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One possible explanation to explain spontaneous DNA fragmentation during in vitro sperm incubation is the involvement of sperm endonuclease activity. We found that there was a significant increase in the amount of DFS in CM (Figure 1). This is in agreement with the described presence of endogenous nucleases in mouse sperm, and it has been reported also in hamster and human spermatozoa (Sotolongo et al, 2005). These authors reported that mammalian spermatozoa can cleave DNA into loop-sized fragments. Recently it was reported by this group that mature mouse spermatozoa contain an active topoisomerase IIB (TOP2B) that regulates DNA degradation in association with an extracellular nuclease (Shaman et al, 2006; Yamauchi et al, 2007). They suggest the possibility that TOP2B and nuclease might be part of an apoptotic mechanism in the sperm cell; another possibility, not mutually exclusive, is that TOP2B can play a role in normal embryogenesis and that the major function of the nuclease may be to serve a protective role by digesting exogenous DNA. Other authors (Maione et al, 1997) also reported that a particular nuclease activity is present in mature sperm. Several studies have pointed out a detrimental effect of long-term incubation on sperm DNA fragmentation. In vitro incubation of swim-up-selected human spermatozoa resulted in a progressive increase in the percentage of DFS (Muratori et al, 2003). Also, an increased percentage of DFS assessed by the sperm chromatin structure assay was found in mammalian sperm incubated in vitro for a long time (Estop et al, 1993). Likewise, we observed an increase in DFS when spermatozoa were incubated for 90 minutes in CM. This effect was only evident when cultures took place in CM because spermatozoa incubated for 90 minutes in M2 medium showed fragmentation values similar to fresh semen. The agents present in CM responsible for this adverse effect could be inactivated because DFS levels returned to values similar to fresh semen when spermatozoa were incubated in CM but in the presence of EDTA. Previous reports have indicated that the presence of EDTA and the absence of Ca2+ and Mg2+ in sperm media improve chromosome stability (Kuretake et al, 1996; Tateno et al, 2000). Sperm DNA damage is prevented by EDTA, confirming that some enzymes requiring Mg2+ and Ca2+, such as endonucleases, may be involved in the DNA degradation mechanism (Szczygiel and Ward, 2002).
The fact that sperm DNA fragmentation did not affect fertilization rates but could affect implantation and postimplantation development has been reported by other authors (Ahmadi and Ng, 1999b; Seli et al, 2004; Tesarik et al, 2004; Zini et al, 2005; Borini et al, 2006). In agreement with these authors, we did not observe a relationship between sperm DNA damage and fertilization rates. Also, we have not found evidence of a deleterious effect of DFS on in vitro embryo development and morphology, in contrast with other studies, in which a negative correlation between sperm DNA fragmentation and blastocyst development after IVF or ICSI has been found (Ahmadi and Ng, 1999b; Seli et al, 2004; Virro et al, 2004; Fatehi et al, 2006). One possible explanation for this discordance might be differences in the source of DNA damage; these authors used x- or gamma-irradiated sperm or sperm from infertile men. However, we have found a negative correlation between the percentage of DFS and the implantation and postimplantation development, indicating a reduced quality of embryos. Even the embryonic genome in mice is activated early in development—the transcription activity of the blastocysts is higher than that detected in the earlier preimplantation stage, and at this stage the paternal genome plays a significant contributory role in embryo function; it is then when consequences of paternal DNA alterations may become manifest, impairing embryo implantation. In concordance with our results, other authors have observed a relationship between DNA fragmentation and implantation (Moskovtsev et al, 2005; Borini et al, 2006). We observed a reduced implantation rate in those groups in which embryos were cultured in vitro to the blastocyst stage, suggesting a negative relationship between ICSI and in vitro culture. This negative effect has been previously reported (Arny et al, 1987; Van der Auwera et al, 1999; Khosla et al, 2001a; Khosla et al, 2001b; Fernández-Gonzalez et al, 2004; Pérez-Crespo et al, 2005), and it questions the convenience of culturing embryos to the blastocyst stage to allow the selection of chromosomally competent embryos that has been proposed by other authors (Sakkas, 1999).
Incubation of the spermatozoa in CM is not a situation that can occur in human fertility clinics; however, a similar mechanism to what we observed can take place when spermatozoa are incubated before injection, especially in samples in which the DFS population is high. It is important to point out that we have observed an increase in the percentage of DFS after only 90 minutes of incubation; this means that long incubations are not necessarily required to increase the DFS percentage in sperm samples. Our data demonstrate that there are factors released from membrane-fragmented spermatozoa capable of inducing DNA fragmentation of intact sperm and therefore significantly reducing the implantation rate and fetal development. Evaluation of spermatozoa DNA fragmentation may prove to be useful to predict implantation rates. If our observations in the mouse can be extrapolated to humans, then our results strongly recommend that ART clinicians should inject sperm without delay to protect the sperm DNA and consequently to decrease the risk of passing damaged information to the offspring, especially if we take into consideration that susceptibility to sperm DNA damage is higher in infertile than in fertile men (Sergerie et al, 2005). Further studies must be carried out to develop techniques that allow us to reduce DNA fragmentation levels in sperm samples used to perform ART.
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Footnotes |
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References |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
Ahmadi A, Ng SC. Fertilizing ability of DNA-damaged spermatozoa. J Exp Zool. 1999b; 284: 696 –704.[CrossRef][Medline]
Aitken RJ. Founders' Lecture. Human spermatozoa: fruits of creation, seeds of doubt. Reprod Fertil Dev. 2004; 16: 655 –664.[Medline]
Aitken RJ, Baker MA. Oxidative stress, sperm survival and fertility control. Mol Cell Endocrinol. 2006; 250: 66 –69.[CrossRef][Medline]
Aitken RJ, Baker MA, Sawyer D. Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic disease. Reprod Biomed Online. 2003; 7: 65 –70.[Medline]
Arnon J, Meirow D, Lewis-Roness H, Ornoy A. Genetic and teratogenic
effects of cancer treatments on gametes and embryos. Hum Reprod
Update. 2001;7: 394
–403.
Arny M, Nachtigall L, Quagliarello J. The effect of preimplantation culture conditions on murine embryo implantation and fetal development. Fertil Steril. 1987; 48: 861 –865.[Medline]
Baker MA, Aitken RJ. Reactive oxygen species in spermatozoa: methods for monitoring and significance for the origins of genetic disease and infertility. Reprod Biol Endocrinol. 2005; 3: 67 .[CrossRef][Medline]
Borini A, Tarozzi N, Bizzaro D, Bonu MA, Fava L, Flamigni C,
Coticchio G. Sperm DNA fragmentation: paternal effect on early
post-implantation embryo development in ART. Hum
Reprod. 2006;21: 2876
–2881.
Braude P, Bolton V, Moore S. Human gene expression first occurs between the four- and eight-cell stages of preimplantation development. Nature. 1988;332: 459 –461.[CrossRef][Medline]
Bungum M, Humaidan P, Spano M, Jepson K, Bungum L, Giwercman A. The
predictive value of sperm chromatin structure assay (SCSA) parameters for the
outcome of intrauterine insemination, IVF and ICSI. Hum
Reprod. 2004;19: 1401
–1408.
Carrell DT, Liu L, Peterson CM, Jones KP, Hatasaka HH, Erickson L, Campbell B. Sperm DNA fragmentation is increased in couples with unexplained recurrent pregnancy loss. Arch Androl. 2003a; 49: 49 –55.[CrossRef][Medline]
Carrell DT, Wilcox AL, Lowy L, Peterson CM, Jones KP, Erickson L, Campbell B, Branch DW, Hatasaka HH. Elevated sperm chromosome aneuploidy and apoptosis in patients with unexplained recurrent pregnancy loss. Obstet Gynecol. 2003b; 101: 1229 –1235.[CrossRef][Medline]
Chan PJ, Corselli JU, Patton WC, Jacobson JD, Chan SR, King A. A simple comet assay for archived sperm correlates DNA fragmentation to reduced hyperactivation and penetration of zona-free hamster oocytes. Fertil Steril. 2001; 75: 186 –192.[CrossRef][Medline]
Estop AM, Munne S, Jost LK, Evenson DP. Studies on sperm chromatin
structure alterations and cytogenetic damage of mouse sperm following in vitro
incubation. Studies on in vitro-incubated mouse sperm. J
Androl. 1993;14: 282
–288.
Fatehi AN, Bevers MM, Schoevers E, Roelen BA, Colenbrander B,
Gadella BM. DNA damage in bovine sperm does not block fertilization and early
embryonic development but induces apoptosis after the first cleavages.
J Androl. 2006;27: 176
–188.
Fernández-Gonzalez R, Moreira P, Bilbao A, Jiménez A,
Pérez-Crespo M, Ramirez MA, Rodriguez De Fonseca F, Pintado B,
Gutiérrez-Adán A. Long-term effect of in vitro culture of mouse
embryos with serum on mRNA expression of imprinting genes, development, and
behavior. Proc Natl Acad Sci U S A. 2004; 101: 5880
–5885.
Filatov MV, Semenova EV, Vorob'eva OA, Leont'eva OA, Drobchenko EA.
Relationship between abnormal sperm chromatin packing and IVF results.
Mol Hum Reprod. 1999; 5: 825
–830.
Garner DL, Johnson LA. Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biol Reprod. 1995; 53: 276 –284.[Abstract]
Genesca A, Caballin MR, Miro R, Benet J, Germa JR, Egozcue J. Repair of human sperm chromosome aberrations in the hamster egg. Hum Genet. 1992; 89: 181 –186.[Medline]
Gutiérrez-Adán A, Granados J, Pintado B, De La Fuente J. Influence of glucose on the sex ratio of bovine IVM/IVF embryos cultured in vitro. Reprod Fertil Dev. 2001; 13: 361 –365.[CrossRef][Medline]
Hales BF, Aguilar-Mahecha A, Robaire B. The stress response in gametes and embryos after paternal chemical exposures. Toxical Appl Pharmacol. 2005;207: 514 –520.[CrossRef][Medline]
Irvine DS, Twigg JP, Gordon EL, Fulton N, Milne PA, Aitken RJ. DNA integrity in human spermatozoa: relationships with semen quality. J Androl. 2000;21: 33 –44.[Abstract]
Ji BT, Shu XO, Linet MS, Zheng W, Wacholder S, Gao YT, Ying DM, Jin
F. Paternal cigarette smoking and the risk of childhood cancer among offspring
of nonsmoking mothers. J Natl Cancer Inst. 1997; 89: 238
–244.
Khosla S, Dean W, Brown D, Reik W, Feil R. Culture of
preimplantation mouse embryos affects fetal development and the expression of
imprinted genes. Biol Reprod. 2001a; 64: 918
–926.
Khosla S, Dean W, Reik W, Feil R. Culture of preimplantation
embryos and its long-term effects on gene expression and phenotype.
Hum Reprod Update. 2001b; 7: 419
–427.
Kodama H, Yamaguchi R, Fukuda J, Kasai H, Tanaka T. Increased oxidative deoxyribonucleic acid damage in the spermatozoa of infertile male patients. Fertil Steril. 1997; 68: 519 –524.[CrossRef][Medline]
Kuretake S, Kimura Y, Hoshi K, Yanagimachi R. Fertilization and development of mouse oocytes injected with isolated sperm heads. Biol Reprod. 1996; 55: 789 –795.[Abstract]
Madrid-Bury N, Pérez-Gutiérrez JF, Pérez-Garnelo S, Moreira P, Pintado Sanjuanbenito B, Gutiérrez-Adán A, de la Fuente Martinez J. Relationship between non-return rate and chromatin condensation of deep frozen bull spermatozoa. Theriogenology. 2005; 64: 232 –241.[CrossRef][Medline]
Maione B, Pittoggi C, Achene L, Lorenzini R, Spadafora C. Activation of endogenous nucleases in mature sperm cells upon interaction with exogenous DNA. DNA Cell Biol. 1997; 16: 1087 –1097.[Medline]
Marchetti F, Wyrobek AJ. Mechanisms and consequences of paternally-transmitted chromosomal abnormalities. Birth Defects Res C Embryo Today. 2005;75: 112 –129.[CrossRef][Medline]
Matsuda Y, Tobari I. Repair capacity of fertilized mouse eggs for x-ray damage induced in sperm and mature oocytes. Mutat Res. 1989;210: 35 –47.[CrossRef][Medline]
McVicar CM, McClure N, Williamson K, Dalzell LH, Lewis SE. Incidence of Fas positivity and deoxyribonucleic acid double-stranded breaks in human ejaculated sperm. Fertil Steril. 2004; 81(suppl 1): 767 –774.[CrossRef][Medline]
Menezo YJ. Paternal and maternal factors in preimplantation embryogenesis: interaction with the biochemical environment. Reprod Biomed Online. 2006;12: 616 –621.[Medline]
Moreira PN, Giraldo P, Cozar P, Pozueta J, Jiménez A,
Montoliu L, Gutiérrez-Adán A. Efficient generation of transgenic
mice with intact yeast artificial chromosomes by intracytoplasmic sperm
injection. Biol Reprod. 2004; 71: 1943
–1947.
Moreira PN, Pérez-Crespo M, Ramirez MA, Pozueta J, Montoliu
L, Gutiérrez-Adán A. Effect of transgene concentration, flanking
matrix attachment regions, and RecA-coating on the efficiency of mouse
transgenesis mediated by intracytoplasmic sperm injection. Biol
Reprod. 2007;76: 336
–343.
Morris ID. Sperm DNA damage and cancer treatment. Int J Androl. 2002;5: 255 –261.
Moskovtsev SI, Willis J, Azad A, Mullen JB. Sperm DNA integrity: correlation with sperm plasma membrane integrity in semen evaluated for male infertility. Arch Androl. 2005; 51: 33 –40.[CrossRef][Medline]
Muratori M, Maggi M, Spinelli S, Filimberti E, Forti G, Baldi E.
Spontaneous DNA fragmentation in swim-up selected human spermatozoa during
long term incubation. J Androl. 2003; 24: 253
–262.
Pérez-Crespo M, Pericuesta E, Rey R, Gutiérrez-Adán A. OC6 Scrotal heat stress in mice affects viability and DNA integrity of sperm, and sex ratio of the offspring. Reprod Domest Anim. 2006; 41(suppl 2): 104 .
Pérez-Crespo M, Ramirez MA, Fernández-González R, Rizos D, Lonergan P, Pintado B, Gutiérrez-Adán A. Differential sensitivity of male and female mouse embryos to oxidative induced heat-stress is mediated by glucose-6-phosphate dehydrogenase gene expression. Mol Reprod Dev. 2005; 72: 502 –510.[CrossRef][Medline]
Sakkas D. The use of blastocyst culture to avoid inheritance of an
abnormal paternal genome after ICSI. Hum Reprod. 1999; 14: 4
–5.
Sakkas D, Urner F, Bizzaro D, Manicardi G, Bianchi PG, Shoukir Y, Campana A. Sperm nuclear DNA damage and altered chromatin structure: effect on fertilization and embryo development. Hum Reprod. 1998; 13(suppl 4): 11 –19.
Saleh RA, Agarwal A, Nada EA, El-Tonsy MH, Sharma RK, Meyer A, Nelson DR, Thomas AJ. Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Fertil Steril. 2003; 79(suppl 3): 1597 –1605.[CrossRef][Medline]
Seli E, Gardner DK, Schoolcraft WB, Moffatt O, Sakkas D. Extent of nuclear DNA damage in ejaculated spermatozoa impacts on blastocyst development after in vitro fertilization. Fertil Steril. 2004; 82: 378 –383.[CrossRef][Medline]
Sergerie M, Laforest G, Bujan L, Bissonnette F, Bleau G. Sperm DNA
fragmentation: threshold value in male fertility. Hum
Reprod. 2005;20: 3446
–3451.
Shaman JA, Prisztoka R, Ward WS. Topoisomerase IIB and an
extracellular nuclease interact to digest sperm DNA in an apoptotic-like
manner. Biol Reprod. 2006; 75: 741
–748.
Shimura T, Inoue M, Taga M, Shiraishi K, Uematsu N, Takei N, Yuan
ZM, Shinohara T, Niwa O. p53-dependent S-phase damage checkpoint and
pronuclear cross talk in mouse zygotes with X-irradiated sperm. Mol
Cell Biol. 2002a;22: 2220
–2228.
Shimura T, Toyoshima M, Taga M, Shiraishi K, Uematsu N, Inoue M, Niwa O. The novel surveillance mechanism of the Trp53-dependent S-phase checkpoint ensures chromosome damage repair and preimplantation-stage development of mouse embryos fertilized with x-irradiated sperm. Radiat Res. 2002b; 158: 735 –742.[CrossRef][Medline]
Sotolongo B, Huang TT, Isenberger E, Ward WS. An endogenous
nuclease in hamster, mouse, and human spermatozoa cleaves DNA into loop-sized
fragments. J Androl. 2005; 26: 272
–280.
Sun JG, Jurisicova A, Casper RF. Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol Reprod. 1997; 56: 602 –607.[Abstract]
Szczygiel MA, Ward WS. Combination of dithiothreitol and detergent
treatment of spermatozoa causes paternal chromosomal damage. Biol
Reprod. 2002;67: 1532
–1537.
Tateno H, Kimura Y, Yanagimachi R. Sonication per se is not as
deleterious to sperm chromosomes as previously inferred. Biol
Reprod. 2000;63: 341
–346.
Tesarik J, Greco E, Mendoza C. Late, but not early, paternal effect
on human embryo development is related to sperm DNA fragmentation.
Hum Reprod. 2004; 19: 611
–615.
Tesarik J, Mendoza C, Greco E. Paternal effects acting during the
first cell cycle of human preimplantation development after ICSI.
Hum Reprod. 2002; 17: 184
–189.
Tomlinson MJ, Moffatt O, Manicardi GC, Bizzaro D, Afnan M, Sakkas
D. Interrelationships between seminal parameters and sperm nuclear DNA damage
before and after density gradient centrifugation: implications for assisted
conception. Hum Reprod. 2001; 16: 2160
–2165.
Van der Auwera I, Pijnenborg R, Koninckx PR. The influence of
in-vitro culture versus stimulated and untreated oviductal environment on
mouse embryo development and implantation. Hum Reprod. 1999; 14: 2570
–2574.
Virro MR, Larson-Cook KL, Evenson DP. Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil Steril. 2004; 81: 1289 –1295.[CrossRef][Medline]
Ward MA, Ward WS. A model for the function of sperm DNA degradation. Reprod Fertil Dev. 2004; 16: 547 –554.[CrossRef][Medline]
Yamauchi Y, Shaman JA, Ward WS. Topoisomerase II-mediated breaks in
spermatozoa cause the specific degradation of paternal DNA in fertilized
oocytes. Biol Reprod. 2007; 76: 666
–672.
Zenzes MT. Smoking and reproduction: gene damage to human gametes
and embryos. Hum Reprod Update. 2000; 6: 122
–131.
Zheng P, Schramm RD, Latham KE. Developmental regulation and in
vitro culture effects on expression of DNA repair and cell cycle checkpoint
control genes in rhesus monkey oocytes and embryos. Biol
Reprod. 2005;72: 1359
–1369.
Zhu BK, Setchell BP. Effects of paternal heat stress on the in vivo development of preimplantation embryos in the mouse. Reprod Nutr Dev. 2004;44: 617 –629.[CrossRef][Medline]
Zini A, Meriano J, Kader K, Jarvi K, Laskin CA, Cadesky K.
Potential adverse effect of sperm DNA damage on embryo quality after ICSI.
Hum Reprod. 2005; 20: 3476
–3480.
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