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JSM Sexual Medicine

Oxidative Stress Effect on the Spermatogenesis Genes Expression in the Mouse Model

Research Article | Open Access

  • 1. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, Iran
  • 2. Prestage Department of Poultry Science, NC State University, USA
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Corresponding Authors
Hamid Gourabi, Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Abstract

We aimed to evaluate the effects of oxidative stress induced by tertiary-butyl hydroperoxide on the DNA Fragmentation Index (DFI) and expression of spermatogenesis genes. Totally, 15 mice of BALB/c strain were categorized into 3 equal groups. To induce oxidative stress (OS), the case group was intraperitoneally injected with 1:10 (Lethal Dose) LD50 (100 μl) of tert-butyl hydroperoxide (t-BHP) for 14 days. Sterile water (200μl) was given intraperitoneally as a placebo in the control group. Whether OS is responsible for gene expression alteration, a third group of mice was treated simultaneously with t-BHP and Taurine for 14 days. In each group, the OS level was determined by the isolation of testicular cells and measurement of H2 O2 and O2 •- levels by flow cytometry. In order to evaluate the incidence of the DFI and apoptosis, tunnel assay was performed on prepared testis tissue samples. By the completion of the treatment phase, the mice were sacrificed and the expression of Dazl, Ddx3y, Smcy and Usp9y genes was measured by RT-PCR in each group. Flow cytometry indicated an increase in reactive oxygen species (ROS) in testicular cells following t-BHP treatment (p < 0.009) and a reduction after Taurine co-administration (p < 0.008). Also, the TUNEL assay showed an increase in DFI in sperm DNA following t-BHP treatment and a reduction after Taurine co-administration (p < 0.008). Our results showed that the ROS decrease (p<0.001) the expression of the mentioned genes and Taurine treatment adjacent to t-BHP significantly reduced the ROS level and protected against downregulation of the Dazl, Ddx3y, Smcy and Usp9y genes (p<0.001, p = 0.030, p = 0.002, p = 0.011 respectively). The oxidative stress may reduce the expression of these Y chromosome genes significantly that are involved in spermatogenesis, and the use of the antioxidant may be a protection against downregulation.

Keywords

• Infertility

• Oxidative stress

• Taurine

• t-BHP

• Gene expression

CITATION

Mokhtari P, Meybodi AM, Fatemi NA, Narenji AG, Gourabi H (2023) Oxidative Stress Effect on the Spermatogenesis Genes Expression in the Mouse Model. JSM Sexual Med 7(3): 1117

INTRODUCTION

The male factor is responsible for 30% of all infertility problems [1]. Oxygen is necessary for the maintenance of normal cellular function [2]. Previous studies have been demonstrated that oxygen metabolite production such as reactive oxygen species (ROS), free radicals and peroxides increase oxygen ions and decrease antioxidants leading to oxidative stress, sperm DNA damage and reduced sperm motility [3]. ROS are known as a reactive chemical species that contain H2 O2 , O2 •- and OH, which can damage intracellular lipids, proteins, and nucleic acids via oxidation-reduction reaction [5]. During normal spermatogenesis, reactive oxygen species are produced. Although, a small level of ROS is needed for capacitation, acrosome reaction and fertilization [5], a high level of ROS may associate with lipid peroxidation and damages of the plasma membrane integrity, genomic expression profile and DNA damages that may lead to a spermatozoa malfunction and infertility [6]. Different factors such as chemotherapy drugs, environmental pollution [7], fertilizers, waste material burners, fossil fuel power plants, and other industrial activities [8] may induce reactive oxygen species in the testis. In addition, diabetes has been reported as an oxidative stress inducer in testis [9].

It has been shown that Hydro peroxides such as t-BHP) can induce significant DNA damages on sperm ad testicular tissue [10]. Moreover, it has been shown that t-BHP induces OS in the other biological systems of mice [11]. The human Y chromosome harbors genes involved in the development of the testis and spermatogenesis are Dazl, Ddx3y, Smcy and Usp9y.

DAZL (Deleted in Azoospermia like), a member of highly conserved DAZ family genes, plays important role in the human fertility due to the control of differentiation, growth, maturation of germ cells and maintain spermatogonia stem cells (SSC) [12] by regulating stage-specific gene expression. Another important gene is DDX3Y, a member of the DEAD-box protein family of ATP dependent RNA helicases, which plays an essential role in the development, maintenance, and fate of early germ cells [13].

Smcy or KDM5D, a Y - link gene, is essential for the spermatogenesis progress that plays an important controlling role in the epigenetic changes during spermatogenesis. Evidence suggests that during the spermatogenesis, SMCY may be involved to some extent in inactivating and concentrating chromosomes before entering meiosis [14]. Another important gene through spermatogenesis procedure is USP9Y, a Y-linked gene encoding the ubiquitin-specific peptidase 9. This testis-specific gene is a major member of the AZFa gene family [15]. This gene encodes a member of the C19 peptidase family and removes ubiquitin residues from ubiquitin-related precursors. It is a vital modulator of a spermatogenesis procedure that plays role in the survival, stability, and protection of germ cells [16]. To our best knowledge, there are two published studies, which reported expression alterations of Dazl and Ddx3y under influence of antibiotics and arsenic and DFI {Seidel, 2019 #38}. Here, for the first time, we investigated the effect of directly induced oxidative stress on the expression of genes involved in the spermatogenesis and DFI before and after treatment in a mouse model.

MATERIALS AND METHODS

This experimental study was approved by the Royan institutions ethical committee of the Research Council, Tehran, Iran (No.: Ec/92/1084). Guidelines of the ethics committee and the declaration of Helsinki were followed for the Care and Use of animals (DHEW publication, NIH, 80–23).

Animals and care

Fifteen adult male mice (Balb/C strain, 8-12 weeks old, 22-25 g) were provided from the Royan mouse stock. One week prior to the experiment initiation, they were kept under controlled condition, including ambient temperature of 25?C ± 3?C, 50% humidity, and an 12:12-hour light/dark cycles. They had free right to use food and water.

Animal treatment

We randomly distributed these 15 mice into 3 equal experimental groups, including: (1) the t-BHP treated group which received t-BHP (Sigma., USA, CAS Number: 75-91-2) at doses equivalent to 1: 10 (LD50) which was determined in our previous study [17] (daily, intraperitoneal (IP) injection of 508 μmol per 100 g/w, for two consecutive weeks. (2)- received a combination of t-BHP (same dose of t-BHP as first group) and Taurine (Sigma Co., USA, CAS Number: 107-35-7) [18], and (3): control group and received only distilled water (200,ip/14 days). Following treatment completion, animals were sacrificed through cervical dislocation and testes were collected.

Evaluation of reactive oxygen species in testes

The induced ROS level of testes tissue samples was measured by flow-cytometry (BD FACS Calibur; Becton-Dickinson, USA) with utilization of the 2’,7’-dichloro-dihydro-fluorescein diacetate (DCFH-DA; Sigma, USA. CAS Number: 4091-99-0) for H2 O2 and di-hydro-ethidium (DHE; Sigma, USA. CAS Number: 104821- 25-2) for O2 •- using a previously described procedure [19,20] with some modifications. Briefly, after enzymatic digestion of testicular tissue by the method of Bellvt and colleagues [19], about 1-3 million testicular cells were homogenized with DCFH-DA (10 mmol) and DHE (1/25 µmol) and then incubated for 15 min at 37 °C in a shaker incubator to allow the probe connected to the membrane-bound vesicles in the darkness. Then the cells were washed several times with PBS (Phosphate Buffer Saline; USA. CAS Number: 21600-010), and the percentage of fluorescent cells was measured by flow cytometry [20]. Green fluorescence (DCF) was evaluated between 500 and 530 nm in the FL-1 channels, and the red fluorescence (HE) was evaluated between 590 and 700 nm in the FL-2 channels.

Sperm Collection and DNA Fragmentation Index (DFI)

Following scarification, the cauda epididymides were dissected and resuspended in a 2 mL of pre-warmed Tissue Culture Medium (TCM -Sigma, USA. CAS Number: M 2520), containing 10% Human serum albumin (HAS; Biotest, Germany. CAS Number: B 05AA01). Mature spermatozoa using swim up, density gradient and simple wash steps were selected and evaluated for the DFI. The DNA fragmentation test was performed by TUNEL assay (Cell Death Detection Kit: Roche, Mannheim, Germany). Briefly, sperm sample (1-3× 106 sperm cells/sample) were washed from seminal plasma by low-speed centrifugation (600 g; 5 minutes), fixed with 4% PBS-buffered for 60 min at room temperature and permeabilized with 0.25% Triton X-100 for 10 min at 4°C. Then the specimen incubated (individually) in a TUNEL reaction mixture in the dark at 37 °C for 1 hour followed by evaluation in a fluorescence microscope. Using a fluorescence microscope (Nikon, Japan), the DFI was calculated as the percentage of Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) positive sperm [21]. Images were captured by a video camera (Basler Vision, A312FC at 50 fps; Tecnologie Co., Ahrensburg, Germany) x 10 magnification.

RNA extraction and cDNA synthesis

The biopsy pieces were frozen immediately using liquid nitrogen. Total RNA was extracted from tissue using the Trizol reagent (Invitrogen Life Technologies; USA. CAS Number: 15596026). RNA samples were quantified by Nano Drop 2000 spectrophotometer (Thermo Scientific, USA). RNA integrity was evaluated by the ratio of 28S/18S ribosomal RNA bands after electrophoresis in a normal 1 % agarose gel. The cDNA was synthesized from 1 μg of the total RNA, treated with deoxyribonuclease I (DnaseI; Fermentase: Gran Island, NY, USA. CAS Number: EN0521) according to the manufacturer protocol (Fermentase: Gran Island, NY, USA, CAS Number: 4368814) and stored at -20 ?C until use.

Quantitative real-time PCR

A primer builds upon existing primer design software for q real time PCR primers, such as Primer3 (Table A2). The β-Actin was gene selected as the housekeeping gene. Quantification of cDNAs was conducted two times by quantitative real timepolymerase chain reaction (qRT-PCR), using the SYBR Green PCR kit (ABI Applied Biosystems, USA. CAS Number: 4309155) and 7500 Real-time PCR System (ABI Applied Biosystems, USA). The reaction conditions were followed by 40 cycles: denaturation at 95°C for 10 min, annealing at 600 C for 1 min. Data were analyzed using the 2ˆ(−ΔΔCt) method [22] (Table A2).

Statistical Analysis

For comparison of the flow cytometry and DFI results between control and treated groups, the data were analyzed by Mann-Whitney (Non parametric) test using SPSS software (SPSS V.16, Inc, Chicago, IL, USA). For comparison of the expression results between control and treated groups, the data were analyzed by One Way ANOVA (supplemented with a Tukey– HSD multiple comparison test) using SPSS software (SPSS V.16, Inc, Chicago, IL, USA).

DISCUSSION

Oxidative stress is one of the male infertility causes [23]. Previously, we showed fourteen days injection of the t-BHP can induce oxidative stress in the mouse testis by a significant increase in H2 O2 and O2 •- levels, which can lead to a significant reduction in adult sperm parameters [5]. Rectifying gene-expression changes and consequently sperm parameters change and also, the sperm DNA fragmentation index, in vivo studies are necessary. Normally, the double-stranded DNA breakage repairs in a short time through homologous recombination. Since the Y chromosome haploid genome contains key spermatogenesis-specific genes and is unable to repair the lost genetic information, therefore it is very sensitive to the deletion of genes and compensating for their reduced expression. It has been observed that increasing the ROS levels may lead to the conversion of guanine to 8-OH guanine (8-OHG), 2 ′-deoxyguanosine and DNA fragmentation, and if not repaired, can cause Y chromosome microdeletions which affect the expression of genes in this region and eventually, cause infertility [24,25].

In this study, to establish a ROS model, after two weeks of peritoneal injection of t-BHP in mice, H2 O2 and O2 •- levels increased significantly in the testes, which proves the induction of oxidative stress. Also, a significant increase in DNA strand breaks was confirmed by comparing case and control groups with the TUNNEL test. A reduction in sperm parameters was also observed. To examine this theory, the alterations of the expression of the specific Ddx3y, Smcy, and Usp9y genes on the mice Y chromosome may cause infertility, a group of genes as mentioned in the introduction, which has high homology with human Y chromosome genes and also Dazl gene on chromosome 3 were tested.

In this study, the downregulation of the Dazl gene was observed after the induction of oxidative stress in the t-BHP treated group, which may lead to a decrease in sperm count. Similarly, in previous studies, two antibiotics, amoxicillin and gentamicin, and also mono-butyl phthalate were associated with increased oxidative stress and sperm morphology, while, decreased sperm motility and the Dazl mRNA expression level [26,27].

Following induction of oxidative stress, a significant decrease in the Ddx3y gene expression was observed, which is probably associated with oligospermia due to a decrease in the mitotic division in germ cells [Table 2]. Deletion of the DDX3Y gene is associated with the complete loss of germ cells and the development of Sertoli cell-only syndrome (SCOS), that leads to azoospermia in humans [28]. Based on a previous study, the arsenic induction of oxidative stress reduces the expression of the Ddx3y gene and sperm count in addition to an increase in the sperm tail and head abnormalities [29].

After induction of oxidative stress in our study, decreased expression of the Smcy gene was observed. Since this gene is involved in the cell cycle controlling and the onset of meiosis [14], a decrease in sperm count [Table 2]. After ROS induction is probably due to this decrease in the Smcy gene expression. In addition, loss or reduction of the SMCY gene expression due to disruption of the regulation of H3K4me3 transcription marker may lead to cell cycle irregularities and increases replication stress. It is possible that the observed increase in the breakdown of double-strand DNA, which remains unrepaired, is associated with a decrease in the expression of this gene, results in the formation of sperm with unrepaired DNA [30] (Table A1).

As mentioned in other studies, spot mutations and deletion of USP9Y can be associated with different phenotypes such as decreased motility (asthenozoospermia), diminished concentration of sperm (oligospermia to azoospermia), and spermatid maturation arrest [31]. In this study, after the induction of oxidative stress, changes in the spermograme were associated with a decrease in Usp9y gene expression.

It has been observed that ROS is neutralized by the enzymatic antioxidant system. Therefore, treatment with antioxidant agents is associated with reducing oxidative stress and has protective effects against ROS-induced damage in the different tissues such as testis. Taurine via NADPH oxidase may alter ROS production through two different ways: first, it may suppress ROS production by mitochondrial electron transfer chains and second: it protects the antioxidant enzymes against oxidative stress damage [32].

After Taurine injection, no significant difference in the expression of specific Dazl, Ddx3y, Smcy, and Usp9y genes were observed in the control group. It seems that the oxidative stress production under the t-BHP injection is deactivated by the Taurine and the normal condition of gene expression and sperm production will return. This also confirms, no other pathways may have influenced the results of our experiments, other than oxidative stress induced by t-BHP.

Although, using inbred mouse rather than outbred, eliminates the effect of background genetic in our animal model, but using human sample will provide more accurate information. We suggest using the Testicular sperm extraction (TESE) samples of those obstructive azoospermia patients with high level of oxidative stress for the next step of this research.

RESULTS

DCFH and DHE processing by the testis cells

The intracellular induction of ROS was detected by flow cytometry using fluorescent probes DCFH-DA and DHE to determine H2 O2 and O2 •- levels. Figure A1 shows

Histogram graphs of flow cytometry of mouse testis stained with  DCFH and DHE. The green line shows development of fluorescent dye of DCFH  which was collected in fluorescence detectors 1 (FL1) and red line shows the  development of fluorescence dye of DHE which was collected in fluorescence  detectors 2 (FL2). (Date analyzed by Mann-Whitney (Non parametric) test). A: shows the level of H2O2 and O2 •- in control group. B: A significant increase  in H2 O2  and O2 •- levels in the t-BHP treated compared to the controls group (p <  0.009). C: A significantly reduced H2O2 and O2 •- in the t-BHP+Taurine treated  compared to the t-BHP group (p < 0.008)

Figure A1: Histogram graphs of flow cytometry of mouse testis stained with DCFH and DHE. The green line shows development of fluorescent dye of DCFH which was collected in fluorescence detectors 1 (FL1) and red line shows the development of fluorescence dye of DHE which was collected in fluorescence detectors 2 (FL2). (Date analyzed by Mann-Whitney (Non parametric) test). A: shows the level of H2O2 and O2 •- in control group. B: A significant increase in H2 O2 and O2 •- levels in the t-BHP treated compared to the controls group (p < 0.009). C: A significantly reduced H2O2 and O2 •- in the t-BHP+Taurine treated compared to the t-BHP group (p < 0.008)

the level of H2 O2 and O2 •- in a mouse selected from each group. A significant increase in H2 O2 and O2 •- level was observed in the t-BHP-treated group in comparison with the control group (p < 0.009). And also, in the t-BHP+Taurine group, the level of H2 O2 and O2 •- was significantly reduced in comparison with the t-BHP group (p < 0.008). The percentage of H2 O2 and O2 •- in the testis was higher in the t-BHP group in comparison with the t-BHP+Taurine group (77%-99% versus 34%-58%) (Figure A2) (p < 0.008).

Status of ROS measured as H2 O2  and O2 •- levels (measured using  DCFH and DHE) in testis treated with t-BHP and t-BHP+Taurine groups  compared to control group by flow-cytometry. (Data analyzed by Mann- Whitney  test, Error bar: STDV)

Figure A2: Status of ROS measured as H2 O2 and O2 •- levels (measured using DCFH and DHE) in testis treated with t-BHP and t-BHP+Taurine groups compared to control group by flow-cytometry. (Data analyzed by Mann- Whitney test, Error bar: STDV)

DFI and TUNNEL Assay

One hundred spermatozoa were counted in duplicate slides. Green- colored sperm cells (TUNEL positive) and red-colored sperm cell (TUNEL negative) were calculated by the counting machine (Figure A3). The percentage of DNA fragmentation in the sperms was higher in the t-BHP group in comparison with the t-BHP+Taurine group (52%-80% versus 16%-23%) (Figure A4) (p < 0.008).

The incidence of sperm DNA damage by TUNNEL staining image:  Image A shows the control group, image B shows the treated group with t-BHP  and image C shows treated group with t-BHP+Taurine). Florescent microscopy.

Figure A3: The incidence of sperm DNA damage by TUNNEL staining image: Image A shows the control group, image B shows the treated group with t-BHP and image C shows treated group with t-BHP+Taurine). Florescent microscopy.

The compression of DNA Fragmentation in the t-BHP and  t-BHP+Taurine groups with the control group by TUNNEL assay. (Data analyzed  by Mann-Whitney test, Error bar: STDV). *** Shows a significant difference at level p < 0.008.

Figure A4: The compression of DNA Fragmentation in the t-BHP and t-BHP+Taurine groups with the control group by TUNNEL assay. (Data analyzed by Mann-Whitney test, Error bar: STDV). *** Shows a significant difference at level p < 0.008.

Gene Expression analysis

qRT-PCR analysis shows that the expression of Dazl, Ddx3y, Smcy, and Usp9y were significantly downregulated in the t-BHP treated group in comparison with the control group (p<0.001). However, the expression of the same genes was significantly upregulated in the t-BHP+Taurine group in comparison with to the t-BHP group (p<0.001, p = 0.030, p = 0.002, p = 0.011 respectively) (Figure A5).

mRNA expression of Dazl, Ddx3y, Smcy, & Usp9y in mice testis. The expression level of Dazl, Ddx3y, Smcy, & Usp9y were significantly  downregulated in the t-BHP treated mice compared to those in the control  group. (Data analyzed by One Way ANOVA Error bar: STDV) (p<0.001). The expression level of Dazl, Ddx3y, Smcy, & Usp9y were significantly  upregulated in the t-BHP+Taurine treated compared to the t-BHP treated. (Data  analyzed by One Way ANOVA Error bar: STDV) Respectively (p<0.001, p =  0.030, p = 0.002, p = 0.011).  * Shows a significant difference at level p<0.05. **Shows a significant difference at level p<0.01. ***Shows a significant difference at level p<0.001.

Figure A5 mRNA expression of Dazl, Ddx3y, Smcy, & Usp9y in mice testis. The expression level of Dazl, Ddx3y, Smcy, & Usp9y were significantly downregulated in the t-BHP treated mice compared to those in the control group. (Data analyzed by One Way ANOVA Error bar: STDV) (p<0.001). The expression level of Dazl, Ddx3y, Smcy, & Usp9y were significantly upregulated in the t-BHP+Taurine treated compared to the t-BHP treated. (Data analyzed by One Way ANOVA Error bar: STDV) Respectively (p<0.001, p = 0.030, p = 0.002, p = 0.011).

* Shows a significant difference at level p<0.05.

**Shows a significant difference at level p<0.01.

***Shows a significant difference at level p<0.001.

CONCLUSION

Oxidative stress can influence gene expression such as Dazl, Ddx3y, Smcy, and Usp9y in the testes and change spermatogenesis in the mouse model. Antioxidants such as Taurine prevent adverse effects of ROS and can be recommended to regulate gene expression and maintain normal spermatogenesis.

FUNDING

This study was funded by a grant from the Royan Institute, Tehran, Iran (grant number 91000256).

ACKNOWLEDGEMENTS

The authors would like to dedicate this paper to the memory of Dr. Saeid Kazemi Ashtiani, the late founder of the Royan Institute. This study was funded by a grant from the Royan Institute, Tehran, Iran and we are thankful of this institutional support.

REFERENCES

1. Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reprod Biol Endocrinol. 2015; 13: 1-9.

2. Agarwal A, Sharma R, Gupta S, Harlev A, Ahmad G, Du Plessis SS, et al. Oxidative stress in human reproduction: shedding light on a complicated phenomenon: Springer; 2017.

3. Wu PY, Scarlata E, O’Flaherty CJA. Long-term adverse effects of oxidative stress on rat epididymis and spermatozoa. 2020;9170.

4. Aitken RJ. Free radicals, lipid peroxidation and sperm function. Reproduction, Fertility and Development. 1995;7:659-68.

5. Fatemi N, Sanati M, Jamali Zavarehei M, Ayat H, Esmaeili V, Golkar? Narenji A, et al. Effect of tertiary?butyl hydroperoxide (TBHP)?induced oxidative stress on mice sperm quality and testis histopathology. Andrologia. 2013;45:232-9.

6. Beckman JK, Coniglio JG. A comparative study of the lipid composition of isolated rat Sertoli and germinal cells. Lipids. 1979; 14: 262-7.

7. 7.Guerriero G, D’Errico G, Di Giaimo R, Rabbito D, Olanrewaju OS, Ciarcia G. Reactive oxygen species and glutathione antioxidants in the testis of the soil biosentinel Podarcis sicula (Rafinesque 1810). Environ Sci Pollut Res Int. 2018; 25: 18286-96.

8. Fasulo S, Guerriero G, Cappello S, Colasanti M, Schettino T, Leonzio C, et al. The “SYSTEMS BIOLOGY” in the study of xenobiotic effects on marine organisms for evaluation of the environmental health status: biotechnological applications for potential recovery strategies. Reviews in Environmental Science and Bio/Technology. 2015; 14: 339-45.

9. Kilarkaje N, Al-Hussaini H, Al-Bader MM. Diabetes-induced DNA damage and apoptosis are associated with poly (ADP ribose) polymerase 1 inhibition in the rat testis. Eur J Pharmacol. 2014; 737: 29-40.

10. Komura K, Yoshikawa Y, Shimamura T, Chakraborty G, Gerke TA, Hinohara K, et al. ATR inhibition controls aggressive prostate tumors deficient in Y-linked histone demethylase KDM5D. J Clin Invest. 2018; 128: 2979-95.

11. Aboua YG, du Plessis SS, Reichgelt P, Brooks N. The in vitro effects of superoxide, some commercially available antioxidants and red palm oil on sperm motility. Asian J Androl. 2009; 11695-702.

12. Zhang Q, Li Q, Li J, Li X, Liu Z, Song D, et al. b-DAZL: A novel gene in bovine spermatogenesis. Progress in Natural Science. 2008; 18: 1209-18.

13. Kotov AA, Olenkina OM, Godneeva BK, Adashev VE, Olenina LV. Progress in understanding the molecular functions of DDX3Y (DBY) in male germ cell development and maintenance. Biosci Trends. 2017: 11: 46-53.

14. Akimoto C, Kitagawa H, Matsumoto T, Kato S. Spermatogenesis? specific association of SMCY and MSH5. Genes to Cells. 2008; 13: 623- 33.

15. Vogt P, Falcao C, Hanstein R, Zimmer J. The AZF proteins. International Journal of Andrology. 2008; 31: 383-94.

16. Ginalski K, Rychlewski L, Baker D, Grishin NV. Protein structure prediction for the male-specific region of the human Y chromosome. Proceedings of the National Academy of Sciences. 2004; 101: 2305-10.

17. Fatemi N, Sanati MH, Jamali Zavarehei M, Ayat H, Esmaeili V, Golkar-Narenji A, et al. Effect of tertiary-butyl hydroperoxide (TBHP)-induced oxidative stress on mice sperm quality and testis histopathology. Andrologia. 2013; 45: 232-9.

18. Ma N, Sasoh M, Kawanishi S, Sugiura H, Piao F. Protection effect of taurine on nitrosative stress in the mice brain with chronic exposure to arsenic. J Biomed Sci. 2010; 17 Suppl 1: S7.

19. Bellve AR, Cavicchia J, Millette CF, O’brien DA, Bhatnagar Y, Dym MJTJocb. Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization. 1977; 74: 68-85.

20. Fatemi N, Sanati MH, Shamsara M, Moayer F, Zavarehei MJ, Pouya A, et al. TBHP-induced oxidative stress alters microRNAs expression in mouse testis. J Assist Reprod Genet. 2014; 31: 1287-93.

21. Hosseinifar H, Sabbaghian M, Chehrazi M, Modarresi T, Alipour FJ, Gilani MAS. Assessment of deoxyribonucleic acid fragmentation index, testicular volume, semen parameters, and hormone profile in gonadotropin-treated men with hypogonadotropic hypogonadism. Urology. 2013; 82: 1291-5.

22. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001; 25: 402-8.

23. Chen H, Liu J, Luo L, Baig MU, Kim JM, Zirkin BR. Vitamin E, aging and Leydig cell steroidogenesis. Exp Gerontol. 2005; 40: 728-36.

24. Kataoka H, Mizuno K, Oda E, Saito A. Determination of the oxidative stress biomarker urinary 8-hydroxy-2’-deoxyguanosine by automated on-line in-tube solid-phasemicroextraction coupled with liquid chromatography- tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2016; 1019: 140-6.

25. Bui A, Sharma R, Henkel R, Agarwal A. Reactive oxygen species impact on sperm DNA and its role in male infertility. Andrologia. 2018; 50: e13012.

26. Karaman M, Budak H, Çiftci M. Amoxicillin and gentamicin antibiotics treatment adversely influence the fertility and morphology through decreasing the Dazl gene expression level and increasing the oxidative stress. Arch Physiol Biochem. 2019; 125: 447-55.

27. Du J, Xiong D, Zhang Q, Li X, Liu X, You H, et al. Mono-butyl phthalateinduced mouse testis injury is associated with oxidative stress and down-regulated expression of Sox9 and Dazl. J Toxicol Sci. 2017; 42: 319-28.

28. Kamp C, Huellen K, Fernandes S, Sousa M, Schlegel P, Mielnik A, et al. High deletion frequency of the complete AZFa sequence in men with Sertoli-cell-only syndrome. Molecular human reproduction. 2001; 7: 987-94.

29. Zeng Q, Yi H, Huang L, An Q, Wang H. Reduced testosterone and Ddx3y expression caused by long-term exposure to arsenic and its effect on spermatogenesis in mice. Environmental toxicology and pharmacology. 2018; 63: 84-91.

30. Kumar TR, Muralidhara. Induction of oxidative stress by organic hydroperoxides in testis and epididymal sperm of rats in vivo. J Androl. 2007; 28: 77-85.

31. Cerván-Martín M, Castilla JA, Palomino-Morales RJ, Carmona FD. Genetic landscape of nonobstructive azoospermia and new perspectives for the clinic. J Clin Med. 2020; 9: 300.

32. Seidel U, Huebbe P, Rimbach G. Taurine: a regulator of cellular redox homeostasis and skeletal muscle function. Mol Nutr Food Res. 2019; 63: 1800569.

Mokhtari P, Meybodi AM, Fatemi NA, Narenji AG, Gourabi H (2023) Oxidative Stress Effect on the Spermatogenesis Genes Expression in the Mouse Model. JSM Sexual Med 7(3): 1117

Received : 23 Aug 2023
Accepted : 30 Sep 2023
Published : 30 Sep 2023
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ISSN : 2573-1548
Launched : 2016
Annals of Emergency Surgery
ISSN : 2573-1017
Launched : 2016
Annals of Mens Health and Wellness
ISSN : 2641-7707
Launched : 2017
Journal of Preventive Medicine and Health Care
ISSN : 2576-0084
Launched : 2018
Journal of Chronic Diseases and Management
ISSN : 2573-1300
Launched : 2016
Annals of Vaccines and Immunization
ISSN : 2378-9379
Launched : 2014
JSM Heart Surgery Cases and Images
ISSN : 2578-3157
Launched : 2016
Annals of Reproductive Medicine and Treatment
ISSN : 2573-1092
Launched : 2016
JSM Brain Science
ISSN : 2573-1289
Launched : 2016
JSM Biomarkers
ISSN : 2578-3815
Launched : 2014
JSM Biology
ISSN : 2475-9392
Launched : 2016
Archives of Stem Cell and Research
ISSN : 2578-3580
Launched : 2014
Annals of Clinical and Medical Microbiology
ISSN : 2578-3629
Launched : 2014
JSM Pediatric Surgery
ISSN : 2578-3149
Launched : 2017
Journal of Memory Disorder and Rehabilitation
ISSN : 2578-319X
Launched : 2016
JSM Tropical Medicine and Research
ISSN : 2578-3165
Launched : 2016
JSM Head and Face Medicine
ISSN : 2578-3793
Launched : 2016
JSM Cardiothoracic Surgery
ISSN : 2573-1297
Launched : 2016
JSM Bone and Joint Diseases
ISSN : 2578-3351
Launched : 2017
JSM Bioavailability and Bioequivalence
ISSN : 2641-7812
Launched : 2017
JSM Atherosclerosis
ISSN : 2573-1270
Launched : 2016
Journal of Genitourinary Disorders
ISSN : 2641-7790
Launched : 2017
Journal of Fractures and Sprains
ISSN : 2578-3831
Launched : 2016
Journal of Autism and Epilepsy
ISSN : 2641-7774
Launched : 2016
Annals of Marine Biology and Research
ISSN : 2573-105X
Launched : 2014
JSM Health Education & Primary Health Care
ISSN : 2578-3777
Launched : 2016
JSM Communication Disorders
ISSN : 2578-3807
Launched : 2016
Annals of Musculoskeletal Disorders
ISSN : 2578-3599
Launched : 2016
Annals of Virology and Research
ISSN : 2573-1122
Launched : 2014
JSM Renal Medicine
ISSN : 2573-1637
Launched : 2016
Journal of Muscle Health
ISSN : 2578-3823
Launched : 2016
JSM Genetics and Genomics
ISSN : 2334-1823
Launched : 2013
JSM Anxiety and Depression
ISSN : 2475-9139
Launched : 2016
Clinical Journal of Heart Diseases
ISSN : 2641-7766
Launched : 2016
Annals of Medicinal Chemistry and Research
ISSN : 2378-9336
Launched : 2014
JSM Pain and Management
ISSN : 2578-3378
Launched : 2016
JSM Women's Health
ISSN : 2578-3696
Launched : 2016
Clinical Research in HIV or AIDS
ISSN : 2374-0094
Launched : 2013
Journal of Endocrinology, Diabetes and Obesity
ISSN : 2333-6692
Launched : 2013
Journal of Substance Abuse and Alcoholism
ISSN : 2373-9363
Launched : 2013
JSM Neurosurgery and Spine
ISSN : 2373-9479
Launched : 2013
Journal of Liver and Clinical Research
ISSN : 2379-0830
Launched : 2014
Journal of Drug Design and Research
ISSN : 2379-089X
Launched : 2014
JSM Clinical Oncology and Research
ISSN : 2373-938X
Launched : 2013
JSM Bioinformatics, Genomics and Proteomics
ISSN : 2576-1102
Launched : 2014
JSM Chemistry
ISSN : 2334-1831
Launched : 2013
Journal of Trauma and Care
ISSN : 2573-1246
Launched : 2014
JSM Surgical Oncology and Research
ISSN : 2578-3688
Launched : 2016
Annals of Food Processing and Preservation
ISSN : 2573-1033
Launched : 2016
Journal of Radiology and Radiation Therapy
ISSN : 2333-7095
Launched : 2013
JSM Physical Medicine and Rehabilitation
ISSN : 2578-3572
Launched : 2016
Annals of Clinical Pathology
ISSN : 2373-9282
Launched : 2013
Annals of Cardiovascular Diseases
ISSN : 2641-7731
Launched : 2016
Journal of Behavior
ISSN : 2576-0076
Launched : 2016
Annals of Clinical and Experimental Metabolism
ISSN : 2572-2492
Launched : 2016
Clinical Research in Infectious Diseases
ISSN : 2379-0636
Launched : 2013
JSM Microbiology
ISSN : 2333-6455
Launched : 2013
Journal of Urology and Research
ISSN : 2379-951X
Launched : 2014
Journal of Family Medicine and Community Health
ISSN : 2379-0547
Launched : 2013
Annals of Pregnancy and Care
ISSN : 2578-336X
Launched : 2017
JSM Cell and Developmental Biology
ISSN : 2379-061X
Launched : 2013
Annals of Aquaculture and Research
ISSN : 2379-0881
Launched : 2014
Clinical Research in Pulmonology
ISSN : 2333-6625
Launched : 2013
Journal of Immunology and Clinical Research
ISSN : 2333-6714
Launched : 2013
Annals of Forensic Research and Analysis
ISSN : 2378-9476
Launched : 2014
JSM Biochemistry and Molecular Biology
ISSN : 2333-7109
Launched : 2013
Annals of Breast Cancer Research
ISSN : 2641-7685
Launched : 2016
Annals of Gerontology and Geriatric Research
ISSN : 2378-9409
Launched : 2014
Journal of Sleep Medicine and Disorders
ISSN : 2379-0822
Launched : 2014
JSM Burns and Trauma
ISSN : 2475-9406
Launched : 2016
Chemical Engineering and Process Techniques
ISSN : 2333-6633
Launched : 2013
Annals of Clinical Cytology and Pathology
ISSN : 2475-9430
Launched : 2014
JSM Allergy and Asthma
ISSN : 2573-1254
Launched : 2016
Journal of Neurological Disorders and Stroke
ISSN : 2334-2307
Launched : 2013
Annals of Sports Medicine and Research
ISSN : 2379-0571
Launched : 2014
Annals of Vascular Medicine and Research
ISSN : 2378-9344
Launched : 2014
JSM Biotechnology and Biomedical Engineering
ISSN : 2333-7117
Launched : 2013
Journal of Hematology and Transfusion
ISSN : 2333-6684
Launched : 2013
JSM Environmental Science and Ecology
ISSN : 2333-7141
Launched : 2013
Journal of Cardiology and Clinical Research
ISSN : 2333-6676
Launched : 2013
JSM Nanotechnology and Nanomedicine
ISSN : 2334-1815
Launched : 2013
Journal of Ear, Nose and Throat Disorders
ISSN : 2475-9473
Launched : 2016
JSM Ophthalmology
ISSN : 2333-6447
Launched : 2013
Journal of Pharmacology and Clinical Toxicology
ISSN : 2333-7079
Launched : 2013
Annals of Psychiatry and Mental Health
ISSN : 2374-0124
Launched : 2013
Medical Journal of Obstetrics and Gynecology
ISSN : 2333-6439
Launched : 2013
Annals of Pediatrics and Child Health
ISSN : 2373-9312
Launched : 2013
JSM Clinical Pharmaceutics
ISSN : 2379-9498
Launched : 2014
JSM Foot and Ankle
ISSN : 2475-9112
Launched : 2016
JSM Alzheimer's Disease and Related Dementia
ISSN : 2378-9565
Launched : 2014
Journal of Addiction Medicine and Therapy
ISSN : 2333-665X
Launched : 2013
Journal of Veterinary Medicine and Research
ISSN : 2378-931X
Launched : 2013
Annals of Public Health and Research
ISSN : 2378-9328
Launched : 2014
Annals of Orthopedics and Rheumatology
ISSN : 2373-9290
Launched : 2013
Journal of Clinical Nephrology and Research
ISSN : 2379-0652
Launched : 2014
Annals of Community Medicine and Practice
ISSN : 2475-9465
Launched : 2014
Annals of Biometrics and Biostatistics
ISSN : 2374-0116
Launched : 2013
JSM Clinical Case Reports
ISSN : 2373-9819
Launched : 2013
Journal of Cancer Biology and Research
ISSN : 2373-9436
Launched : 2013
Journal of Surgery and Transplantation Science
ISSN : 2379-0911
Launched : 2013
Journal of Dermatology and Clinical Research
ISSN : 2373-9371
Launched : 2013
JSM Gastroenterology and Hepatology
ISSN : 2373-9487
Launched : 2013
Annals of Nursing and Practice
ISSN : 2379-9501
Launched : 2014
JSM Dentistry
ISSN : 2333-7133
Launched : 2013
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