Testes Of Male Mice Research Articles

Effects of low-dose rate radiation on immune and epigenetic regulation of the mouse testes

Abstract This study investigated the effects of chronic low-dose-rate radiation exposure on gene expression related to immunological defense mechanisms and epigenetic regulation in the testis of male mice. The study found that radiation exposure (4.5 mGy/hr until 4000 mGy) led to a significant decrease in the expression of genes involved in epigenetic regulation and a significant increase in the expression of several genes linked to innate and acquired immunity, suggesting a noticeable effect on the immune response. In addition, there was a significant increase in the expression of SRY-box transcription factor 9, which plays a crucial role in testicular development, indicating a response to irradiation. These findings provide insights into the biological effects of chronic low-dose-rate radiation exposure on innate immunity and epigenetic regulation in the testis.

PWWP3A deficiency accelerates testicular senescence in aged mice.

The PWWP domain-containing proteins are involved in chromatin-associated biological processes, including transcriptional regulation and DNA repair, and most of them are significant for gametogenesis and early embryonic development in mammals. PWWP3A, one of the PWWP domain proteins, is a reader of H3K36me2/H3K36me3 and a response factor to DNA damage. However, the physiological role of PWWP3A in spermatogenesis and fertility remains unclear. The goal of this study was to explore the function and mechanism of PWWP3A in the process of spermatogenesis. We generated V5-Pwwp3a KI mice and PWWP3A polyclonal antibody to observe the localization of PWWP3A in vivo. Meanwhile, Pwwp3a KO mice was used to explore the function in spermatogenesis. We reported that PWWP3A is a predominant expression in the testis of mice. During spermatogenesis, PWWP3A exhibits the temporal expression from early-pachytene to the round spermatids. The results of spermatocyte spreading and immunostaining showed that PWWP3A aggregated on the XY body, which then diffused as the XY chromosome separated at late-diplotene. Although the depletion of PWWP3A had no obvious reproductive defects in young male mice, there were observed morphological abnormalities in sperm heads. Immunoprecipitation demonstrated the interaction of PWWP3A with DNA repair proteins SMC5/6; however, PWWP3A deficiency did not result in any meiotic defects. Notably, the testes of aged male Pwwp3a KO mice displayed pronounced degeneration, and were characterized by the presence of vacuolated seminiferous tubules. Furthermore, RNA-seq analysis revealed an upregulation in the expression of genes which may be involving in immunoregulatory and inflammatory response pathways in aged Pwwp3a KO mice with testicular degeneration. Our study showed that PWWP3A was highly enriched in the mouse testis, and the Pwwp3a KO mice were fertile. However, the aged Pwwp3a KO male mice displayed testicular atrophy that may be due to changes in the immune micro-environment or abnormal repair of DNA damage.

Effect of Ethanolic Extract of Avocado (Persea americana Mill.) Seed on the Testis Histology (Mus musculus l.) Induced Monosodium Glutamate (MSG)

Monosodium Glutamate (MSG) is a substance that is able to produce free radicals such as ROS (Reactive Oxygen Species), which can cause oxidative stress in the body and damage reproductive organs such as the testes. The purpose of this study was to determine the effect of avocado seed ethanol extract (Persea americana Mill.) on the histology of the testes of mice (Mus musculus L.) induced monosodium glutamate (MSG). This research is an experimental study that was conducted in October 2023 – January 2024 at Biology Laboratory, Faculty of Mathematics and Natural Sciences, Universitas Negeri Padang. The design used is a Complete Randomized Design (CRD) with 4 treatments and 6 repeats. The data obtained were analyzed using the ANOVA test (Analysis of variance) using the SPSS 23 application. The results showed that the administration of avocado seed ethanol extract showed a noticeable effect on the diameter of the testes of male mice exposed to MSG. However, it showed no noticeable influence on testicular weight, seminiferous tubule diameter and Leydig cell count. Ethanol extract avocado seed dose 0.84 mg /g body weight / day (P1) was able to improve the diameter of mice testes that had been induced MSGKeyword: Avocado seed extract, Histology of testis, Mice, MSG

Orally Administered Silver Nanoparticles Are Absorbed and Migrate to Testes in Mice.

Given that daily exposure to nanoparticles is now unavoidable, there are concerns that nanoparticles have unexpected biological effects due to their small size. Here, we examined the biodistribution of silver nanoparticles, which are the most frequently used nanoparticles owing to their antibacterial activity, with a diameter of 10 nm (nAg10) to the male genital tract, and the effects of paternal treatment with nAg10 on fetal development. Male Slc:ICR male mice were orally treated with nAg10 for 14 consecutive days. Inductively coupled plasma mass spectrometry analysis detected silver in the blood and testis of male mice, but no general toxicological effects were induced. Moreover, there were no significant changes in fetal development when these treated male mice were mated with nontreated female mice. This implies that although orally ingested nAg10 is distributed to the male genital tract, it does not affect fetal development under the present treatment conditions.

Impact of chronic sleep deprivation on male reproductive health: Insights from testicular and epididymal responses in mice.

Sleep deprivation (SD) can cause damage to the male reproductive system. However, the duration required for such damage and the specific sequence and severity of damage to the testis and epididymis remain unclear. To investigate the effects of different durations of SD on different parts of the testis and epididymis caput, corpus, and cauda. Adult ICR mice were randomly assigned to five groups: the SD group (SD for 18h/day for 1, 2, 3, or 4weeks), the SD + Vit E group (supplemented with Vit E 50mg/kg/d during 4weeks of SD, the SD+NS group (saline supplementation during 4weeks of SD), the SD + RS group (5weeks of recovery sleep after 4weeks of SD), and a normal sleep control (Ctrl) group. Following the interventions, sperm parameters, testicular and epididymal histopathology, inflammatory response, and oxidative stress markers were compared between the groups. Compared to the Ctrl group, the SD group showed a decrease in sperm motility and concentration from SD 2W and SD 3W, respectively. Decreases in sperm concentration and motility were more pronounced in the cauda compared to the caput and corpus. Pathological damage was less severe in the epididymis caput than in the corpus and cauda. After 4weeks of SD, inflammation and oxidative stress increased in both testes and epididymis. Both sleep recovery and vitamin E supplementation showed significant improvements, though they did not fully reach the level of the Ctrl group. Chronic SD for more than 2weeks causes varying degrees of damage to the testis, epididymis caput, corpus, and cauda in male mice. This damage is not fully reversible after 5weeks of sleep recovery and antioxidant stress treatment. These findings help us to identify and prevent SD damage to the male reproduction at an early stage.

Differential changes in end organ immune cells and inflammation in salt-sensitive hypertension: effects of increasing M2 macrophages.

Salt-sensitive hypertension (SSHTN) is associated with M1 macrophage polarization and inflammatory responses, leading to inflammation-associated lymphangiogenesis and functional impairment across multiple organs, including kidneys and gonads. However, it remains unclear whether promoting M2 macrophage polarization can alleviate the hypertension, inflammation, and end organ damage in mice with salt sensitive hypertension (SSHTN). Male and female mice were made hypertensive by administering nitro-L-arginine methyl ester hydrochloride (L-NAME; 0.5 mg/ml) for 2 weeks in the drinking water, followed by a 2-week interval without any treatments, and a subsequent high salt diet for 3 weeks (SSHTN). AVE0991 (AVE) was intraperitoneally administered concurrently with the high salt diet. Control mice were provided standard diet and tap water. AVE treatment significantly attenuated BP and inflammation in mice with SSHTN. Notably, AVE promoted M2 macrophage polarization, decreased pro-inflammatory immune cell populations, and improved function in renal and gonadal tissues of mice with SSHTN. Additionally, AVE decreased lymphangiogenesis in the kidneys and testes of male SSHTN mice and the ovaries of female SSHTN mice. These findings highlight the effectiveness of AVE in mitigating SSHTN-induced elevated BP, inflammation, and end organ damage by promoting M2 macrophage polarization and suppressing pro-inflammatory immune responses. Targeting macrophage polarization emerges as a promising therapeutic approach for alleviating inflammation and organ damage in SSHTN. Further studies are warranted to elucidate the precise mechanisms underlying AVE-mediated effects and to assess its clinical potential in managing SSHTN.

Evaluation of the Potential Protective Role of Galangin Associated with Gold Nanoparticles in the Histological and Functional Structure of Testes of Adult Male Albino Mice Administrated with Carbon Tetrachloride

Galangin (Gal) is a natural flavonoid sourced from the roots of the plant Albina Kalanga. It possesses a variety of pharmacological activities, such as antioxidant, anticancer, and anti-inflammatory. The current study aims to enhance the efficacy of galangin through the use of gold nanoparticles as a drug delivery system against CCl4-induced toxicity in the testicular tissue of male albino mice. Forty-two albino mice were divided into seven groups (6 mice/group), treatment with carbon tetrachloride solution for two weeks by intraperitoneal injection, (1 ml/kg) once a week for all groups except the control group, after which a group was injected with gold nanoparticles and two groups were injected with galangin in two concentrations, and two groups were injected with AuNPs + Gal conjugation solution with two concentrations. The animals' bodies were weighed and blood samples were obtained for testosterone hormone analysis and testicles for the purpose of weighing and histological study at the end of the experiments. The study showed that the testicular tissue of mice treated with CCl4 had a different pathology compared to the control group, the testosterone hormone levels in the CCl4 group were significantly higher than those of the control group. Results from the AuNPs+Gal group showed a significant reduction in the effects of CCl4 toxicity on mice testicles, with testosterone levels are returning to normal and histological testicle structures improving, the ratio of testicle weights to animal body weight in the group injected with AuNPs was significantly higher than that of the control and CCl4 groups. In conclusion, the results revealed that galangin combined with gold nanoparticles could effectively reduced the histological and functional tissue damages caused by CCl4, presenting a promising natural solution that needs to be further developed

Insulin improves spermatogenesis in a mouse model of aging

Aging can have adverse effects on the chance of fertility in men. In fact, men over 40 years old usually suffer from low sperm quality, sperm motility, and low level of serum testosterone that leads to infertility. In addition, the structure of DNA is more vulnerable in aged testes due to the high level of reactive oxygen species (ROS) and nitric oxide. However, it has been showed that insulin is able to decrease oxidative stress and apoptosis. Moreover, some studies showed that insulin can enhance both spermatogenesis and fertility. In this regard, we investigate the impacts of insulin on spermatogenesis, the number of sperm, and the level of gonadal hormones in aged testes of mice. Thirty-six adult male NMRI mice were used. They were kept under standard conditions. The animals were divided into two groups control and insulin. The insulin group received 100 µL insulin with a 72-hour interval by intraperitoneal (IP) injection for 33 weeks and the control group received water. At the 8, 12, and 33 weeks of treatment, the testes of mice were removed for histological, cellular (ROS and glutathione [GSH]) and molecular (Bcal2, caspase-3, and Bax) analysis. For measurement of the level of hormones (testosterone, LH, and FSH), the blood samples were collected from the heart. According to our findings, insulin could significantly increase the volume of the testes and the total number of spermatogonial stem cells, primary spermatocytes, round spermatids as well as leydig cells. In addition, activity of GPX was considerably high in the insulin group. Furthermore, in the insulin group, the level of ROS was reduced. In conclusion, insulin may be effective in enhancing the function of the testes in aged male mice.

Bisphenol Analogs Downregulate the Self-Renewal Potential of Spermatogonial Stem Cells.

In this study, we investigated the effect of bisphenol-A (BPA) and its major analogs, bisphenol-F (BPF), and bisphenol-S (BPS), on spermatogonial stem cells (SSCs) populations using in vitro SSC culture and in vivo transplantation models. SSCs enriched from 6- to 8-day-old C57BL/6-eGFP+ male mice testes were treated with varying concentrations of bisphenols for 7 days to examine bisphenol-derived cytotoxicity and changes in SSC characteristics. We utilized flow cytometry, immunocytochemistry, real-time quantitative reverse transcription-PCR, and western blot analysis. The functional alteration of SSCs was further investigated by examining donor SSC-derived spermatogenesis evaluation through in vivo transplantation and subsequent testis analysis. BPF exhibited a similar inhibitory effect on SSCs as BPA, demonstrating a significant decrease in SSC survival, inhibition of proliferation, and induction of apoptosis. On the other hand, while BPS was comparatively weaker than BPA and BPF, it still showed significant SSC cytotoxicity. Importantly, SSCs exposed to BPA, BPF, and BPS exhibited a significant reduction in donor SSC-derived germ cell colonies per total number of cultured cells, indicating that, like BPA, BPF, and BPS can induce a comparable reduction in functional SSCs in the recipient animals. However, the progress of spermatogenesis, as evidenced by histochemistry and the expressions of PCNA and SSC specific markers, collectively indicates that BPA, BPF, and BPS may not adversely affect the spermatogenesis. Our findings indicate that the major BPA substitutes, BPF and BPS, have significant cytotoxic effects on SSCs, similar to BPA. These effects may lead to a reduction in the functional self-renewal stem cell population and potential impacts on male fertility.

The Effect of Moringa oleifera Extract on Histopathological of Mice Testes Exposed by Monosodium Glutamate

Background: Monosodium glutamate (MSG) consumption in excess can have an impact on a man's fertility and lead to infertility. MSG has the potential to produce free radicals, which can harm cells. Antioxidants can fight off free radicals. Exogenous antioxidants are required to maintain equilibrium since there will be an imbalance if the body produces more free radicals than endogenous antioxidants. Exogenous antioxidants flavonoids are present in the leaves of the Moringa plant (Moringa oleifera L). It is thought that flavonoids have 4-5 times more antioxidant potential than vitamins. Objectives: To evaluate how moringa leaf extract affects the histopathological profile of MSG-exposed mouse testes. Methods: Only the posttest was used as a control group in this study. 25 male mice were used as the sample, and they were split up into five groups. The treatment group 1 (P1) received 6 g/day of MSG exposure + 300 mg/kg BW/day of moringa leaf extract, the treatment group 2 (P2) received 6 g/day of MSG exposure + 600 mg/kg BW/day of moringa leaf extract, and the treatment group 3 (P3) received 6 g/day of MSG exposure + 1200 mg/kg BW/day of moringa leaf extract. The control group (K-) received only standard feed. The mice were put to death after 30 days. The testes were removed for histological examination using hematoxylin-eosin staining, and the Johnsen scoring criteria were applied to their evaluation. Results: The data were not significant according to the Shapiro-Wilk test in the P3 group (MSG 6 g/day + Moringa Extract 1200 mg/kg BW/day). The Kruskal-Wallis test revealed that there was no significant difference between the seminiferous tubules according to the treatment group (p = 0.117). Conclusion: Moringa oleifera leaf extract protects the histopathological picture of the testes of male mice compared to the group given monosodium glutamate.

Withaferin-A attenuates diabetes mellitus induced male reproductive dysfunction mediated by ERα in brain and testes of Swiss albino mice

Diabetes mellitus (DM) is a chronic metabolic disease, characterized by persistent hyperglycemia resulting from diminished insulin secretion or insulin resistance. The present study evaluated the ameliorative effects of Withaferin-A (WA) on DM-induced reproductive dysfunction in mice. For the same, mice were intraperitoneally injected with Streptozotocin (STZ), (40 mg/kg/day) for 5 consecutive days to induce DM. Mice were then treated with WA (8 mg/kg/day) in normal and diabetic conditions (STZ + WA). Next, blood glucose levels, oral glucose tolerance, intraperitoneal insulin tolerance, oxidative stress and reproductive parameters were estimated. For reproductive performance, immunofluorescent localization of gonadotropin-releasing hormone (GnRH-I) and estrogen receptor alpha (ERα) in the preoptic area and paraventricular nucleus region of hypothalamus and ERα in testes was performed. STZ-induced diabetes triggered reproductive dysfunctions as mediated by low GnRH-I and ERα in the brain and ERα in the testes along with declined testosterone and estradiol levels. Treatment with WA significantly reduced the blood glucose levels and enhanced glucose clearance accompanied by reduced oxidative stress in the brain, pancreas and testes as indicated by the low levels of H2O2 and MDA in diabetic mice treated with WA (STZ + WA). This study reports, for the first time, that WA can efficiently ameliorate DM-induced reproductive dysfunctions by enhancing endogenous testosterone, estrogen and increased GnRH-I and ERα in the brain and ERα in the testes of DM-induced male mice.

Catalysis-Mediated Male Contraception through Black Phosphorus Nanosheets.

Nanocontraception has been proposed and received extensive attention in recent years for population control. However, currently developed methods for nanocontraception still face problems in efficacy and safety. Here, we propose catalysis-mediated oxidation as a new strategy for nanocontraception. With the catalytic production of highly oxidative species, male contraception was successfully achieved after the administration of black phosphorus nanosheets into the testes of male mice. Further mechanistic studies revealed that contraception was induced by oxidative stress and apoptosis of spermatogenesis cells. Meanwhile, the apoptosis of germ cells released testis antigen and induced immune cell infiltration, which enhanced reproductive damage. Notably, the introduced black phosphorus nanosheets naturally degraded during the catalytic oxidation process and ultimately converted to harmless phosphates, indicating the safety of the strategy. Furthermore, the catalysis-mediated strategy avoids utilizing additional inducers, such as near-infrared irradiation, magnetic fields, or ultrasound, which may cause severe pain. In summary, the proposed catalysis-mediated contraception can be a self-cleared, convenient, and safe strategy for controlling male fertility.

Melatonin protects Leydig cells from HT-2 toxin-induced ferroptosis and apoptosis via glucose-6-phosphate dehydrogenase/glutathione -dependent pathway.

HT-2 toxin is a mycotoxin commonly found in food and water that can have adverse effects on male reproductive systems, including testosterone secretion. Ferroptosis and apoptosis are two types of programmed cell death that have been implicated in the regulation of cellular functions. Melatonin, a powerful antioxidant with various physiological functions, has been shown to regulate testosterone secretion. However, the mechanisms underlying the protective effects of melatonin against HT-2 toxin-induced damage in testosterone secretion are not fully understood. In this study, we investigated the effects of HT-2 toxin on sheep Leydig cells and the potential protective role of melatonin. We found that HT-2 toxin inhibited cell proliferation and testosterone secretion of Leydig cells in a dose-dependent manner and induced ferroptosis and apoptosis through intracellular reactive oxygen species accumulation, leading to lipid peroxidation. Exposure of Leydig cells to melatonin in vitro reversed the defective phenotypes caused by HT-2 toxin via a glucose-6-phosphate dehydrogenase/glutathione-dependent mechanism. Interference of glucose-6-phosphate dehydrogenase disrupted the beneficial effect of melatonin on ferroptosis and apoptosis in HT-2 toxin-treated Leydig cells. Furthermore, similar results were observed in vivo in the testes of male mice injected with HT-2 toxin with or without melatonin treatment for 30 days. Our findings suggest that melatonin inhibits ferroptosis and apoptosis by elevating the expression of glucose-6-phosphate dehydrogenase to eliminate reactive oxygen species accumulation in HT-2 toxin-treated Leydig cells. These results provide fundamental evidence for eliminating the adverse effects of HT-2 toxin on male reproduction.

Obesity induces male mice infertility via oxidative stress, apoptosis, and glycolysis.

The current declining trend in male fertility parallels the increasing prevalence of obesity worldwide. This paper revealed that the poor in vitro fertilization rates and decreased sperm motility in obese mice due to excessive oxidative stress enhanced apoptosis and impaired glucose metabolism in the testes. Obesity is an urgent public health problem in recent decades, linked to reduced reproductive potential, and negatively affects the success of assisted reproduction technology. The aim of this study is to investigate the mechanisms underlying impaired male fertility caused by obesity. Male C57BL/6 mice fed a high-fat diet for 20 weeks served as mouse models with moderate (20% < body fat rate (BFR) < 30%) and severe obesity (BFR > 30%). Our results showed poor in vitro fertilization rates and decreased sperm motility in obese mice. Abnormal testicular structures were identified in male mice with moderate and severe obesity. The expression level of malondialdehyde increased with obesity severity. This finding indicates that oxidative stress plays a role in male infertility caused by obesity, which was further confirmed by the decreased expression of nuclear factor erythroid 2-related factor 2, superoxide dismutase, and glutathione peroxidases. Our study also found that the expression of cleaved caspase-3 and B-cell lymphoma-2 showed an obesity severity-dependent manner indicating that apoptosis is highly correlated with male infertility caused by obesity. Moreover, the expression of glycolysis-related proteins, including glucose transporter 8, lactate dehydrogenase A, monocarboxylate transporter 2 (MCT2), and MCT4, decreased significantly in the testes of obese male mice, suggesting energy supply for spermatogenesis is impaired by obesity. Taken together, our findings provide evidence that obesity impairs male fertility through oxidative stress, apoptosis, and blockage of energy supply in the testes and suggest that male obesity influences fertility through complex and multiple mechanisms.

Effects of Dioxin on DNA Damage in the Testes of Adult Male Mice

Background: Given the importance of environmental factors and the impact of environmental pollutants, such as dioxins, on organ systems, especially the reproductive system, it is necessary to study these issues and the mechanisms of their effects. Objectives: This study aimed to investigate the effects of different doses of dioxins on DNA damage in the testes of adult male mice. Methods: In this experimental study, 32 male Naval Medical Research Institute (NMRI) rats were randomly divided into four groups: The control group received normal saline, and the dioxin groups were treated with different doses (0.1, 0.5, and 1 µg/kg) for two weeks. Apoptosis in the testes was then examined using a TUNEL assay kit. Results: The mean number of TUNEL-positive spermatogonia cells was 5.91±5.28 in the dioxin group 1, 7.20±10.03 in the dioxin group 2, and 8.73±4.63 in the dioxin group 3, which was higher than that in the control group (0.16±0.40; P=0.073, P=0.034, and P=0.007, respectively). The mean number of TUNEL-positive spermatocyte cells was 5.16±1.99 in the dioxin group 1, 2.50±4.62 in the dioxin group 2, and 3.33±2.94 in the dioxin group 3, which was higher than that in the control group (P=0.034, P=0.14, and P=0.037, respectively). The mean number of TUNEL-positive spermatocyte cells in the dioxin group 1 was significantly higher than that in the dioxin group 2 (2.50±4.62, P=0.047). The average number of TUNEL-positive spermatid cells was 11.58±6.90 in the dioxin group 1, 11.10±12.19 in the dioxin group 2, and 10.20±7.32 in the 3-dioxin group, which was higher than that in the control group (0.16±0.40; P=0.008, P=0.014, and P=0.015, respectively). Conclusion: The results of the present study showed that dioxin caused dose-dependent apoptosis in the testes.

Evaluation of hazards of electronic -cigarette’s liquid refill on testes of mice, complemented by histopathological and chromatographic analysis

ABSTRACT Electronic-cigarettes (e-cigarettes) are devices designed to become an alternative to classic cigarettes. Vaping of e-cigarettes and their recharge liquid have become extremely popular among the adolescents; however, its safety is not well established. Evaluation of the components of e-cigarette liquid and their potential effects on testis of adult male mice. This aim will be fulfilled by histological, ultrastructural, and immunohistochemical analysis of mice testis biopsies. Twenty mice were allocated into two groups of equal size. The control group was given regular saline, whereas the treated group was given e-liquid (contains 3 mg of nicotine/kg of body weight) both groups daily intraperitoneally injected for 3 weeks. Analysis of e-liquid by Gas Chromatography-Mass Spectrometric GC/MS demonstrated nicotine, phenol, vanillin, aldehydes, and pyrethroid insecticide. Evaluation of oxidative stress parameters revealed significant reduction of SOD and GPx. Histological results revealed a significant reduction in the height of seminiferous tubules, sloughing of spermatogenic cells, most cells being dark and pyknotic, and thickening of the interstitium with accumulation of PAS positive exudate. Most spermatogenic cells showed degenerative changes as rarefied cytoplasm, ill-defined electron-dense nuclei, and elongated spermatid showed deformity of ectoplasmic specialization. Immunohistochemical studies revealed a significant increase in caspase-3 positive cells and a significant reduction of area % of E-cadherin. The analysis of an available E-liquid demonstrated potentially harmful chemicals that are not shown in the labeling of the product. E-liquid appears to impair anti-oxidant defense and cause degenerative changes in the body and disruption of blood testes barrier BTB. So, e-cigarettes cannot be regarded as a non-harmful smoking replacement.

The effect of co-administration of artemisinin and N-acetyl cysteine on antioxidant status, spermatological parameters and histopathology of testis in adult male mice.

This invivo study aimed to evaluate the effect of various concentrations of artemisinin (Art) alone or together with N-acetyl cysteine (NAC) on spermatological indices, antioxidant status, and histopathological parameters of testicular tissue in adult male mice. Six groups of five healthy male mice (25-30g) were randomly assigned to different experimental groups. These groups received DMSO and corn oil (0.1%) as an Art solvent (Control), 50mgkg-1 Art (Art-50), 250mgkg-1 Art (Art-250), 50mgkg-1 Art+150mgkg-1 NAC (Art-50+NAC-150), 250mgkg-1 Art+150mgkg-1 NAC (Art-250+NAC-150) and 150mgkg-1 NAC (NAC-150) for a period of 7days. Testes and epididymis were prepared to evaluate the malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), spermatological indices, and histological parameters. We showed that the high dose of Art (Art-250) significantly reduced the sperm count, motility, viability, and the activity of CAT and increased the levels of MDA compared to the control group. Also, the overdose of Art caused adverse changes in testicular tissue. Co-administration of NAC with Art (Art-250+NAC-150) corrected the adverse effects of Art. The current study reports that a high dose of Art affects, spermatological parameters, antioxidant/stress oxidative status of the male reproductive system, and NAC is capable neutralize all adverse effects caused by Art.

Dietary selenium excess affected spermatogenesis via DNA damage and telomere-related cell senescence and apoptosis in mice.

Selenium (Se) is a vital microelement for spermatogenesis and male fertility. The aim of this study was to investigate the effects of Se on the male reproductive function and possible mechanisms. Fourty male mice were randomly divided into 0, 0.1, 0.3 and 0.9mg/kg Se supplementation groups and given with Se dietary intervention for 12 weeks. Our data showed that excessive Se intake damaged the tissue structure of testes and epididymides of the mice, resulting in decreased sperm quality and quantity. Moreover, excessive Se induced oxidative stress, causing DNA damage and activated DNA damage repair factors (Mre11/Rad50/Nbs1), and also disrupted telomere function by shortening telomere length and decreasing TERT expression. Se excess activated the senescence pathway p53/p21/p16, leading to germ cell senescence, and inhibited cell proliferation by suppressing the Sirt1/Foxo1/c-Myc pathway. All of this led to spermatogenic cell apoptosis, thereby causing a decrease of sperm quantity and quality. In conclusion, excessive Se caused reproductive toxicity via inducing telomere dysfunction due to DNA damage, leading to germ cellular senescence and apoptosis in the testes of male mice. Our research provide new proof to explain the underlying mechanism of male reproductive toxicity triggered by excessive Se intake.

Lhb−/−Lhr−/− Double Mutant Mice Phenocopy Lhb−/− or Lhr−/− Single Mutants and Display Defects in Leydig Cells and Steroidogenesis

In the mouse, two distinct populations of Leydig cells arise during testis development. Fetal Leydig cells arise from a stem cell population and produce T required for masculinization. It is debated whether they persist in the adult testis. A second adult Leydig stem cell population gives rise to progenitor-immature-mature adult type Leydig cells that produce T in response to LH to maintain spermatogenesis. In testis of adult null male mice lacking either only LH (Lhb-/-) or LHR (Lhr-/-), mature Leydig cells are absent but fetal Leydig cells persist. Thus, it is not clear whether other ligands signal via LHRs in Lhb null mice or LH signals via other receptors in the absence of LHR in Lhr null mice. Moreover, it is not clear whether truncated LHR isoforms generated from the same Lhr gene promoter encode functionally relevant LH receptors. To determine the in vivo roles of LH-LHR signaling pathway in the Leydig cell lineage, we generated double null mutant mice lacking both LH Ligand and all forms of LHR. Phenotypic analysis indicated testis morpho-histological characteristics are identical among double null and single mutants which all showed poorly developed interstitium with a reduction in Leydig cell number and absence of late stage spermatids. Gene expression analyses confirmed that the majority of the T biosynthesis pathway enzyme-encoding mRNAs expressed in Leydig cells were all suppressed. Expression of thrombospondin-2, a fetal Leydig cell marker gene was upregulated in single and double null mutants indicating that fetal Leydig cells originate and develop independent of LH-LHR signaling pathway in vivo. Serum and intratesticular T levels were similarly suppressed in single and double mutants. Consequently, expression of AR-regulated genes in Sertoli and germ cells were similarly affected in single and double mutants without any evidence of any additive effect in the combined absence of both LH and LHR. Our studies unequivocally provide genetic evidence that in the mouse testis, fetal Leydig cells do not require LH-LHR signaling pathway and a one-to-one LH ligand-LHR signaling pathway exists in vivo to regulate adult Leydig cell lineage and spermatogenesis.

Hadh deficiency induced oligoasthenoteratozoospermia through the TNF-α/Bcl-2 pathway in male mice.

The process of spermatogenesis is a complex and delicate process that is still not fully understood. In this study, we examined the role of fatty acid oxidase 3-hydroxy acyl CoA dehydrogenase (HADH) in maintaining normal spermatogenesis in mice. In male mice, ablation of the Hadh gene using CRISPR/Cas9 technology arrested spermatocyte meiosis, increased multinucleated giant germ cells and vacuoles in seminiferous tubules, and accompanied with acrosomal dysplasia. Hadh-/- male mice showed the typical features of oligoasthenoteratozoospermia (OAT), including decreased sperm concentration and motility and increased sperm abnormalities. Next, we explored the molecular events in the testes of the mutant mice. We found fatty acids accumulated in the testis of Hadh-/- mice. And also, inflammatory factors TNF-α, IL-1β, and IL-6 were significantly increased, apoptosis-related protein Bcl-2 was decreased, and Bax and cleaved-Caspase3 were increased in Hadh-/- male mice testis. After using etanercept, a specific inhibitor of TNF-α, testis injury caused by Hadh knockout was significantly alleviated, the sperm quality and motility were improved, and germ cell apoptosis was reduced. So our study demonstrated that Hadh deletion caused an increase in fatty acids. The accumulated fatty acids further induced testicular inflammation and germ cell apoptosis through the TNF-α/Bcl-2 signaling pathway, finally resulting in OAT in the Hadh-/- mice. Inhibiting TNF-α may be used as a new treatment approach for testicular inflammation and OAT.