Effects of different doses of ethanol on sperm parameters, chromatin structure and apoptosis in adult mice

Rahimipour, Marzieh; Talebi, Ali Reza; Anvari, Morteza; Sarcheshmeh, Abolghasem Abbasi; Omidi, Mansoor

doi:10.1016/j.ejogrb.2013.06.038

Cited by 46 publications

(30 citation statements)

References 48 publications

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“…TEM (Figure 1D(a–f)) and in comparison to control germ cells (a), showed the typical features of germ cell apoptosis in ETRs as chromatin condensation (b,c,e), nuclear pyknosis (d), and late degradation within SCs after phagocytosis (f), in keeping with earlier reports [23,32]. As germ cell apoptosis may be induced by oxidative-nitrosative stress in SCs and/or androgen suppression [23,24,25,26,27], the authors investigated the expression of iNOS and AR protein levels. The 24-h time point was chosen for analysis in subsequent experiments because it showed the highest expression of iNOS and autophagy proteins in the testes of ETRs as reported below, in keeping with a recent study [12].…”

Section: Resultssupporting

confidence: 88%

“…Excessive ethanol consumption in humans and experimental animals has been reported to enhance germ cell apoptosis, inducing infertility problems via mechanisms related to oxidative stress, upregulation of inducible nitric oxide synthase (iNOS) and inflammatory cytokines, DNA damage, androgen suppression and mitochondrial dysfunction [8,23,24,25,26,27,28,29]. However, in the studies that highlighted these effects, SCs were resistant to apoptosis.…”

Section: Introductionmentioning

confidence: 99%

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Upregulated Autophagy in Sertoli Cells of Ethanol-Treated Rats Is Associated with Induction of Inducible Nitric Oxide Synthase (iNOS), Androgen Receptor Suppression and Germ Cell Apoptosis

Horibe

Eid

Ito

et al. 2017

IJMS

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This study was conducted to investigate the autophagic response of Sertoli cells (SCs) to acute ethanol toxicity using in vivo and in vitro models. Adult Wistar rats were intraperitoneally injected with either 5 g/kg ethanol or phosphate-buffered saline (for the control group) and sacrificed 0, 3, 6 and 24 h after injection. Compared to the control group, enhanced germ cell apoptosis was observed in the ethanol-treated rats (ETRs) in association with upregulation of iNOS and reduced expression of androgen receptor protein levels in SCs, which were resistant to apoptosis. Meanwhile, autophagy was upregulated in ETR SCs (peaking at 24 h) compared to the control group, as evidenced by transcription factor EB (TFEB) nuclear translocation, enhanced expression of microtubule-associated protein 1 light chain3-II (LC3-II), lysosome-associated membrane protein-2 (LAMP-2), pan cathepsin protein levels and reduced expression of p62. This upregulation of SC autophagy was confirmed ultrastructurally by enhanced formation of autophagic vacuoles and by immunofluorescent double labelling of autophagosomal and lysosomal markers. Study of cultured SCs confirmed enhanced autophagic response to ethanol toxicity, which was cytoprotective based on decreased viability of SCs upon blocking autophagy with 3-methyladenine (3-MA). The results highlighted the molecular mechanisms of prosurvival autophagy in ETR SCs for the first time, and may have significant implications for male fertility.

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Section: Resultssupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Upregulated Autophagy in Sertoli Cells of Ethanol-Treated Rats Is Associated with Induction of Inducible Nitric Oxide Synthase (iNOS), Androgen Receptor Suppression and Germ Cell Apoptosis

Horibe

Eid

Ito

et al. 2017

IJMS

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“…Ethanol abuse results in the production of spermatozoa with less condensed chromatin, and this may be one possible cause of infertility following ethanol consumption [23]. It is reported that alcohol has negative effects on sperm parameters, chromatin/DNA integrity, and apoptosis in mice [24]. From human studies, it is known that alcohol consumption produces significant morphological changes in the spermatozoa, which include breakage of the sperm head, distention of the midsection, and tail curling [22, 55].…”

Section: Discussionmentioning

confidence: 99%

“…Alcohol exposure causes alterations of the endocrine system controlling the hypothalamic-pituitary-testicular axis function and a direct toxic effect on testis and/or male accessory glands [20–22]. Alcohol exposure is also reported to influence sperm DNA integrity [23, 24]. …”

Section: Introductionmentioning

confidence: 99%

Clinical Factors Associated with Sperm DNA Fragmentation in Male Patients with Infertility

Komiya

Kato

Kawauchi

et al. 2014

The Scientific World Journal

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Objective. The clinical factors associated with sperm DNA fragmentation (SDF) were investigated in male patients with infertility. Materials and Methods. Fifty-four ejaculates from infertile Japanese males were used. Thirty-three and twenty-one were from the patients with varicoceles and idiopathic causes of infertility, respectively. We performed blood tests, including the serum sex hormone levels, and conventional and computer-assisted semen analyses. The sperm nuclear vacuolization (SNV) was evaluated using a high-magnification microscope. The SDF was evaluated using the sperm chromatin dispersion test (SCDt) to determine the SDF index (SDFI). The SDFI was compared with semen parameters and other clinical variables, including lifestyle factors. Results. The SDFI was 41.3 ± 22.2% (mean ± standard deviation) and did not depend on the cause of infertility. Chronic alcohol use increased the SDFI to 49.6 ± 23.3% compared with 33.9 ± 18.0% in nondrinkers. The SDFI was related to adverse conventional semen parameters and sperm motion characteristics and correlated with the serum FSH level. The SNV showed a tendency to increase with the SDFI. The multivariate analysis revealed that the sperm progressive motility and chronic alcohol use were significant predictors of the SDF. Conclusion. The SCDt should be offered to chronic alcohol users and those with decreased sperm progressive motility.

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“…The alkaline comet assay is being widely used as a standard test method since it detects DNA damage including single strand breaks, double strand breaks and akali labile site (15). The comet assay has been used widely to evaluate testicular and sperm toxicity including measurement of ROS, antioxidant enzymes and apoptosis (16–21). …”

Section: Introductionmentioning

confidence: 99%

Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells

Jeon

Kim

et al. 2017

ToxicolRes

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Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration (IC50) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

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Effects of different doses of ethanol on sperm parameters, chromatin structure and apoptosis in adult mice

Cited by 46 publications

References 48 publications

Upregulated Autophagy in Sertoli Cells of Ethanol-Treated Rats Is Associated with Induction of Inducible Nitric Oxide Synthase (iNOS), Androgen Receptor Suppression and Germ Cell Apoptosis

Upregulated Autophagy in Sertoli Cells of Ethanol-Treated Rats Is Associated with Induction of Inducible Nitric Oxide Synthase (iNOS), Androgen Receptor Suppression and Germ Cell Apoptosis

Clinical Factors Associated with Sperm DNA Fragmentation in Male Patients with Infertility

Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells

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