Tsga10 expression correlates with sperm profiles in the adult formalin‐exposed mice

Testis‐specific gene antigen10 (Tsga10), as a cytoskeletal protein in the sperm tail, impacts the sperm motility. This study investigates the correlation between sperm profile alterations and Tsga10 gene expression in adult mice exposed to formaldehyde (FA) and then treated with antioxidant effect of manganese (Mn²⁺). In this regard, we examined 35 NMRI adult male mice (6–8 weeks age) in 4 groups of control, sham, FA‐exposed and FA+Mn²⁺. The mice in FA+Mn²⁺ group were exposed to FA (10 mg kg^?1 twice a day) for 2 weeks and treated with daily Mn²⁺ administration (5 mg kg^?1) in the second week prior to sacrificing the mice for testis dissection. The right testis was dissected in each group and subjected to RNA extraction and cDNA syntheses for gene expression analysis by real‐time PCR. The findings revealed that FA decreased sperm parameters and Tsga10 expression (52.6 ± 24.37%). However, the injected powerful manganese antioxidant improved sperm profile through overexpression of Tsga10 (121.6 ± 27.13%) under FA‐induced stressful condition which proves the correlation between sperm profile and Tsga10 expression (P ≤ 0.05). This study also shows that Tsga10 expression protects sperm dysfunction in FA+Mn²⁺ group and resulting in better preservation of spermatozoa and improvement of male fertility.     

Evaluation of the protective effect of quercetin against cisplatin‐induced renal and testis tissue damage and sperm parameters in rats

The protective effect of quercetin on cisplatin‐induced renal and testicular tissue damage was investigated using biochemical, histopathological and histological approaches. A total of 40 male rats were divided into 5 groups as follows: control; cisplatin alone; quercetin alone; cisplatin + quercetin; and quercetin + cisplatin. Cisplatin was administered to rats at a single dose of 7 mg kg^?1 intraperitoneal. Quercetin was administered by gavage daily for 10 days at dosage 50 mg kg^?1. At the end of the study serum, total antioxidant capacity (TAC) levels and total oxidant status (TOS) were determined. Malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and xanthine oxidase (XO) were studied separately in serum, renal tissue and testicular tissue. Renal and testicular morphological alterations were assessed, histopathologically. Epididymal sperm concentration, motility and morphology were investigated. Testicular and renal TAC and TOS values did not alter significantly. Renal CAT levels were increased by cisplatin and cisplatin plus quercetin groups that is reversed by administration of quercetin before cisplatin. MDA, CAT, SOD ve XO levels of testicular tissue did not differ significantly. Cisplatin and cisplatin plus quercetin groups had decreased sperm motility ratio and increased abnormal spermatozoa. Quercetin partially reverses some of the cisplatin‐related pathological effects on kidney and testis.     

Impact of cigarette‐smoking on sperm DNA methylation and its effect on sperm parameters

DNA methylation is an epigenetic modification of the genome. The purpose of this study was to determine the influence of cigarette‐smoking on sperm DNA methylation from a genomewide survey of sperm samples and to ascertain its effect on sperm parameters. Twenty‐eight sperm DNA samples (from 14 fertile smokers as a case study and 14 proven fertile nonsmokers as controls) were subjected to Infinium 450K BeadChip arrays to identify the changes in the DNA methylation level between the two groups. Then, deep bisulphite sequencing was used to validate five CpGs on 78 samples. The results from the Infinium 450K found that only 11 CpGs showed a significant difference in DNA methylation between the case and the control groups. Five CpGs of the eleven (cg00648582, cg0932376, cg19169023, cg23841288 and cg27391564) underwent deep bisulphite sequencing where cg00648582, related to the PGAM5 gene, and the cg23841288 CpGs, related to the PTPRN2 gene amplicons, showed a significant increase in their DNA methylation level in more than one CpG in the case group. In contrast, a significant decrease was found at cg19169023 and at its various neighbouring CpGs in the TYRO3 gene‐related amplicons. Furthermore, this study demonstrated a significant correlation between the variation in sperm DNA methylation level and standard sperm parameters in the case group.     

Effect of aerobic exercise,low‐fat and high‐fat diet on the testis tissue and sperm parameters in obese and nonobese mice model

Semen quality and male fertility depend on numerous factors such as age, environment, lifestyle, physical activity, genetic background and occupation. We aimed to access the effect of aerobic exercise, low‐ and high‐fat diet on mice testis tissue, and sperm function. Obese and nonobese male mice C57BL/6 were exposed to high fat (Hf) or low fat (Lf) and/or activity (Exe: exercise or Sed: sedentary). Finally, testicular morphometric characteristics, sperm concentration and motility (light microscopy), sperm morphology (eosin/nigrosin dye), lipid peroxidation (BODIPY C11 Probe), chromatin (acridine orange and chromomycin A3 staining) were compared within obese groups (Hf/Exe, Lf/Exe, Lf/Sed, Hf/Sed) and nonobese groups (Hf/Exe, Lf/Exe, Lf/Sed, Hf/Sed). Both exercise and diet interventions did not show any alteration in testicular morphological characteristics, sperm morphology and DNA fragmentation within both obese and nonobese groups (p > 0.05). Exercise and/or diet resulted in a significant increase in sperm concentration and motility within both groups (p < 0.05). Exercise in both groups leads to high percentage of lipid peroxidation (p < 0.05). Exercise intervention significantly improved sperm protamine deficiency within obese group (p < 0.05). We concluded that exercise intervention was more effective than diet in improvement of sperm function within obese groups.     

Application of erythrocyte lysing buffer (ELB) has detrimental effects on human sperm quality parameters,DNA fragmentation and chromatin structure

Erythrocyte lysing buffer (ELB) facilitates the search for spermatozoa by eliminating erythrocytes in testicular suspension used during the ICSI procedure. This study investigates the effects of ELB on sperm quality parameters, sperm chromatin and sperm DNA fragmentation. Normal ejaculations were used as the model for testicular spermatozoa in this study. After swim-up, the sperm pellets were divided into two parts. Part I, the control (Group A), was diluted with culture media; and Part II, the intervention group (Group B), was diluted with ELB for 10 min. After centrifugation in both groups, the sperm pellets were re-suspended with culture media. The samples were immediately evaluated (A0 and B0) and then evaluated again after 1 hr (A1 and B1). The results indicated ELB decreased the progressive motility (81.60 ± 8.69 vs. 64.69 ± 19.08) and viability (97.62 ± 3.02 vs. 85.91 ± 11.46), in Group A and B, respectively, both immediately and 1 hr after preparation. Also, ELB engendered a significant increase in the DNA fragmentation index both immediately (9.68 ± 3.55 vs. 14.38 ± 6.52) and after 1 hr (10.37 ± 5.03 vs. 19.38 ± 6.39). In conclusion, ELB may damage sperm cells, shown by a decreased motility and viability, and it increased DNA fragmentation. Therefore, the use of ELB in testicular semen handling should be discouraged.     

Low sperm recovery from the undescended testis with testicular sperm extraction in postpubertal cryptorchids: preliminary report

As postpubertal cryptorchid testes are generally atrophic and have high malignancy risk, orchiectomy is recommended as the treatment method. However, a few case reports advocated orchiopexy due to finding sperm afterwards. The present clinical study aimed at determining the possibility of finding spermatozoa in postpubertal cryptorchid testis with testicular sperm extraction (TESE). Initially testicular biopsy and later TESE was performed in the undescended testis of 22 unilateral postpubertal cryptorchids and in one of the testes of three bilateral postpubertal cryptorchids. Histopathological assessment of testicular biopsy specimen showed that three patients had maturation arrest and 22 had seminiferous tubular atrophy. By TESE, spermatozoa were found in only one specimen of 25 testes. Our results show that the expectancy to find spermatozoa in postpubertal cryptorchid testis is extremely low by current sperm recovery methods and orchiectomy is still the accurate method of treatment.     

Onco‐testicular sperm extraction: testicular sperm extraction in azoospermic and very severely oligozoospermic cancer patients

An increased risk of testicular cancer in men with infertility and poor semen quality has been reported. In view of the high cure rates for testicular germ cell tumours, increasing clinical importance is being placed on the protection of fertility. High‐dose cytostatic therapy may be expected to cause long‐term infertility. Thus, the standard procedure for fertility protection is the cryopreservation of ejaculated spermatozoa or testicular tissue before therapy. Four male patients with azoospermia and two patients with very severe oligozoospermia underwent onco‐testicular sperm extraction (TESE). We attempted onco‐TESE in patients with azoospermia and very severe oligozoospermia after orchiectomy. Of the patients with testicular germ cell tumours, four had spermatozoa in their testicular tissues. Sertoli cell‐only syndrome was found in one patient, and one patient showed maturation arrest without the detection of spermatozoa. Three of six showed seminomatous germ cell tumour, two of six had nonseminomatous germ cell tumour and one patient showed no malignancy. Two patients achieved clinical pregnancy. Fertility challenges in men with cancer are the most straightforward because of the relative ease of obtaining and cryopreserving sperm. Testicular sperm extraction is a useful technique for obtaining spermatozoa before cytotoxic therapy in azoospermic and very severely oligozoospermic cancer patients.     

Recovery effect of pre‐germinated brown rice on the alteration of sperm quality,testicular structure and androgen receptor expression in rat model of depression

Depression and antidepressant drugs induce adverse effects in male reproduction. Therefore, it is important to investigate alternative treatment for depression without adverse effects on the male reproductive system. The aim of this study was to determine the effect of pre‐germinated brown rice (PGBR) on sperm quality, testicular structure and androgen receptor (AR) expression in rat model of depression. Male Sprague Dawley rats were divided into five groups including control (distilled water only), depression induced by forced swimming test (FST), FST + fluoxetine (antidepressant drug), FST + GABA (gamma‐aminobutyric acid) (standard) and FST + PGBR. When compared with the control, sperm motility showed a significant decrease in FST + fluoxetine group. Sperm morphology also decreased significantly in depression and FST + fluoxetine groups. The morphological changes of seminiferous tubules showed significant increases in depression and FST + fluoxetine groups, while AR expression showed significant decreases in depression, FST + fluoxetine and FST + GABA groups. Interestingly, there were no significant differences in all sperm quality parameters, testicular structure and AR expression in FST + PGBR group. These findings reflect the recovery effects of PGBR treatment on sperm quality, morphological changes of seminiferous tubules and AR expression in stress‐induced rats. Therefore, PGBR may potentially develop for the treatment for depression without adverse effect on male reproduction.     

Studies on relationship between testicular capsule and sperm transport in rat testis

Aim: In SD rats, histological changes in the testis were observed after bilateral capsulotomy (of the tunica albuginea) in order to investigate the physiological role of the testicular capsule on sperm transport. Methods: Bilateral longitudinal capsulotomy was devised to disrupt the capsular contractile function. With this technique, only the tunica vaginalis and tunlca albuginea were slit open, leaving the tunica vasculosa intact to embrace the underlying testicular parenchyma. After capsulotomy, the structural changes in the seminiferous tubules, the transitional distal seminiferous segment, and the rete testis were observed. Results: In the capsulotomized testis, there was sperm retention at the transitional seminiferous segment and progressive degenerative changes in seminiferous tubules. Conclusion: The results clearly indicated that an intact testicular capsule was required for normal sperm transport from the seminiferous tubules into the rete testis. This is the first attempt to study the physiological role of the testicular capsule in intact animals.     

Double‐blind,randomised, placebo‐controlled trial on the effect of L‐carnitine and L‐acetylcarnitine on sperm parameters in men with idiopathic oligoasthenozoospermia

Carnitine is essential for energy metabolism and spermatozoa maturation. Combining L‐carnitine and L‐acetylcarnitine with micronutrients has been investigated as a treatment for infertility in men. We evaluated the effects of a therapeutic formulation, Proxeed Plus, on sperm parameters in oligoasthenozoospermic men. This prospective, randomised, double‐blind, placebo‐controlled clinical trial involved 175 males (19–44 years) with idiopathic oligoasthenozoospermia who failed to impregnate their partners (12 months). Males received Proxeed Plus or placebo for 3 and 6 months. Sperm volume, progressive motility and vitality significantly (p < 0.001) improved after 6 months compared to baseline. Sperm DNA fragmentation index significantly decreased compared to baseline (p < 0.001) and the 3‐month therapy (p = 0.014) in treated men. Increased seminal carnitine and α‐glucosidase concentration also positively correlated with improved progressive motility. Decreased DNA fragmentation index was the good predictor of progressive sperm motility >10%, and simultaneous measurement of changes in sperm vitality and DNA fragmentation index gave the highest probability of sperm motility 10% (AUC = 0.924; 95% CI = 0.852–0.996; p < 0.001). Logistic regression analyses revealed DNA fragmentation index decrease as the only independent predictor of sperm motility 10% (OR = 1.106; p = 0.034). We have demonstrated the beneficial effects of carnitine derivatives on progressive motility, vitality and sperm DNA fragmentation. Combining metabolic and micronutritive factors is beneficial for male infertility.     

Apoptotic sperm biomarkers and the correlation between conventional sperm parameters and clinical characteristics

The principal aim of this retrospective study was to examine the relationship between sperm apoptotic biomarkers and the patient's biclinical characteristics, the conventional sperm parameters and the results of assisted reproductive technology. Sperm analysis, activated caspases, annexin V staining for phosphatidylserine (PS) externalisation and labelling assay for DNA fragmentation were assessed in 122 males of infertile couples. Fifty‐seven couples were allocated to the natural conception group, and 65 couples underwent IVF or ICSI. Semen of IVF/ICSI patients showed a higher proportion of apoptotic spermatozoa in their spermatozoa when compared with a natural conception group (p < .05). Sperm apoptotic biomarkers correlated with age, FSH, and conventional sperm parameters. DNA fragmentation correlated positively with the percentage of semen having externalised PS (r = .78, p = 0) and activated caspases (r = .71, p = 0). Patients without clinical pregnancy had higher frequency of DNA fragmentation, externalised PS and activated caspases compared to patients with clinical pregnancy (p < .001). The best specificity and greater sensitivity were obtained with the test of the DNA fragmentation compared to the other biomarkers. Among the apoptotic biomarkers, only DNA fragmentation was found to predict natural or assisted pregnancy better than conventional sperm parameters.     

The effect of cysteine and glutamine on human sperm functional parameters during vitrification

Assuming the adverse effects of reactive oxygen species (ROS) on sperm function, this study was conducted to assess the effects of cysteine and glutamine as effective antioxidants on human sperm parameters under vitrification. Twenty normozoospermic samples were used. The samples were subjected to a vitrification process and cysteine (5 and 10 mM) and glutamine (10 and 15 mM). The sperm motility parameters, mitochondrial membrane potential (MMP), plasma membrane integrity (PMI), DNA damage and intracellular ROS damage were assessed for each sample. Statistical analyses showed that motility, mitochondrial membrane potential and DNA damage decreased in the vitrified groups with cysteine 5, 10 mM and glutamine 10, 15 mM separately. Also intracellular ROS increased significantly compared to the fresh group (p < .05). No significant differences were observed for PMI compared with the fresh group (p > .05). Supplementation of cysteine and glutamine in both concentrations separately decreased intracellular ROS and DNA damage of spermatozoa with significant increase in PMI, MMP and progressive motility compared to vitrified control group (p < .05). The results showed no significant effect of a specific concentration in cysteine and glutamine on sperm parameters compared to other concentrations. Both amino acids have the potential to improve the harmful effects of freezing on sperm parameters.     

Antioxidant supplementations in vitro improve rat sperm parameters and enhance antioxidant enzyme activities against dimethoate-induced sperm damages

Organophosphorus compounds are currently among the most frequently used pesticides worldwide, and therefore, the potential for human exposure to man is considerable. Their toxicity results in negative effects on many organs and systems such as the male reproductive system. So, vitamins that can offer spermatozoa protection are of great importance. This study was designed to investigate (i) the possibility of dimethoate, an organophosphate insecticide, to induce oxidative stress response in rat spermatozoa in vitro and its effect on antioxidant defence system and (ii) the role of vitamin C and vitamin E in alleviating the cytotoxic effects of dimethoate Epididymal spermatozoa were incubated for 3 h at 37 °C with different concentrations of dimethoate (50, 100 and 200 μm) without vitamins or pre-incubated with 20 mm of vitamin C or 2 mm of vitamin E. Sperm parameters, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels were performed. Dimethoate caused a significant induction of oxidative damage in spermatozoa at different concentrations as evidenced by increased MDA levels. However, a significant decrease in sperm mobility, viability and activities SOD, CAT and GPx was observed. Vitamins pre-treated spermatozoa showed a significant protection against the cytotoxic effects induced by dimethoate on studied parameters.     

Effect of cysteine and glutamine added to extender on post‐thaw sperm functional parameters of buffalo bull

Amino acids seem to be crucial components for semen freezing extender due to antioxidant properties. Therefore, this study aimed to assess motility parameters, membrane integrity, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and DNA damage to detect the optimum concentrations of cysteine and glutamine for buffalo semen cryopreservation. Twenty ejaculates of four buffalo bulls were diluted in tris‐egg yolk extender and divided into seven equal groups consisting of cysteine (5, 7.5 and 10 mmol), glutamine (10, 15 and 20 mmol) and no additive. Supplementation of 5 and 7.5 mmol cysteine and 15 mmol glutamine in cryopreservation extender significantly increased post‐thaw motility and plasma membrane integrity of spermatozoa with significant reduction in intracellular ROS when compared with control groups (P < 0.05). Cysteine at 7.5 mmol concentration elevated progressive motility and MMP, compared with control (P < 0.05). No significant differences were observed for motion patterns and DNA damage of frozen–thawed buffalo spermatozoa in extender containing amino acids. The findings of this study showed that supplementation of 7.5 mmol cysteine and 15 mmol glutamine in semen cryopreservation extender has more potential to decrease intracellular ROS, and subsequently elevate motility and membrane integrity of buffalo frozen–thawed spermatozoa.     

Additional deleterious effects of alcohol consumption on sperm parameters and DNA integrity in diabetic mice

The aim of this study was to survey the impact of alcohol consumption on sperm parameters and DNA integrity in experimentally induced diabetic mice. A total of 32 adult male mice were divided into four groups: mice of group 1 served as control fed on basal diet, group 2 received streptozotocin (STZ) (200 mg kg^?1, single dose, intraperitoneal) and basal diet, group 3 received alcohol (10 mg kg^?1, water soluble) and basal diet, and group 4 received STZ and alcohol for 35 days. The cauda epididymidis of each mouse was dissected and placed in 1 ml of pre‐warm Ham's F10 culture medium for 30 min. The swim‐out spermatozoa were analysed for count, motility, morphology and viability. Sperm chromatin quality was evaluated with aniline blue, toluidine blue, acridine orange and chromomycin A3 staining. The results showed that all sperm parameters had significant differences (P < 0.05), also when sperm chromatin was assessed with cytochemical tests. There were significant differences (P < 0.001) between the groups. According to our results, alcohol and diabetes can cause abnormalities in sperm parameters and chromatin quality. In addition, alcohol consumption in diabetic mice can intensify sperm chromatin/DNA damage.     

Effects of obesity on sperm retrieval,early embryo quality and clinical outcomes in men with nonobstructive azoospermia undergoing testicular sperm aspiration‐intracytoplasmic sperm injection cycles

The objective of this study was to assess the effects of body mass index (BMI) on sperm retrieval, early embryo quality and clinical outcomes in patients with nonobstructive azoospermia (NOA) undergoing testicular sperm aspiration‐intracytoplasmic sperm injection (TESA‐ICSI). A total of 3,005 infertile couples were evaluated between January 2010 and June 2017, including 1585 normal‐weight (BMI < 25 kg/m²), 847 overweight (BMI 25–29.99 kg/m²) and 573 obese (BMI ≥ 30 kg/m²) patients. We found no significant relationship between BMI and sperm retrieval rate (22.4%, 24.3% and 25.1%, p = 0.327) or sperm motility. Among the 705 patients with NOA who underwent TESA‐ICSI cycles, obese individuals had lower T levels and higher E2 levels than normal‐weight and overweight individuals. However, there were no significant differences in other male hormones (follicle stimulating hormone [FSH], luteinizing hormone [LH], or prolactin [PRL]) among the groups. We also found that the sperm parameters, embryo quality and clinical outcomes of patients with NOA undergoing TESA‐ICSI were not influenced by high BMI levels. In conclusion, this study demonstrated a lack of obvious effects of obesity on sperm retrieval, early embryo quality and clinical outcomes in infertile men undergoing TESA‐ICSI cycles, although T and E2 levels were affected.     

Protective effect of Pumpkin seed extract on sperm characteristics,biochemical parameters and epididymal histology in adult male rats treated with Cyclophosphamide

Cancer treatment with cyclophosphamide (CP) may result in reproductive toxicity as one of its side effects. The pumpkin seed is a rich natural source of antioxidant. We have assessed the possible protective efficacy of pumpkin seed extract on sperm characteristics, biochemical parameters and epididymal histology of CP‐treated rats. Male adult Wistar rats were categorised into four groups. Group 1 served as control and received intraperitoneal (IP) injection of isotonic saline solution. Group 2 rats were treated with CP by IP injection in a single dose of 100 mg/kg body weight, only once. Group 3 and 4 received CP plus 300 and 600 mg/kg pumpkin seed extract respectively. Six weeks after treatment, sperm characteristics, biochemical parameters and histopathological changes were examined. Results showed that, sperm characteristics in CP‐treated rats were significantly decreased. Biochemical analysis results showed that the co‐administration of 300 mg pumpkin seed extract could increase the total antioxidant capacity (TAC) level significantly. In CP‐treated rats, histopathological changes such as vacuolisation, disorganisation and separation of epididymal epithelium were observed as well. Interestingly, pumpkin seed extract could improve the above‐mentioned parameters remarkably in CP‐treated rats. Our findings indicated that pumpkin seed extract might be used as protective agent against CP‐induced reproductive toxicity.     

Therapeutic effect of pentoxifylline on reproductive parameters in diabetic male mice

In this study, we aimed to investigate the effects of pentoxifylline (PTX) on male reproductive parameters in diabetic mice. Male adult mice (n = 24) were divided into control and three experimental groups (n = 6) including Diabetic, Diabetic + PTX and PTX groups. Diabetes was induced by single injection of streptozotocin (60 mg kg^?1). PTX was administered intraperitoneally at the dose of 12 mg kg^?1 for 14 days 1 week after diabetes induction. Serum levels of testosterone and blood glucose were determined and collected spermatozoa from cauda epididymidis analysed. Based on histological slides prepared from testis, the diameter of seminiferous tubules was determined using Motic camera and software and also apoptosis using TUNEL assay. Data were analysed using one‐way anova method, and P < 0.05 was considered statistically significant. The mean of seminiferous tubules diameter, final body weight, testis weight, sperm parameters and testosterone hormone level in PTX‐treated diabetic group indicated a significant increase compared to diabetic one, whereas apoptosis index and blood glucose were decreased in this comparison (P < 0.05). These results suggest that intraperitoneal administration of PTX is a potentially beneficial agent to reduce testicular damage and improves sperm parameters in diabetic mice by decreasing the ratio of apoptosis.     

In vitro effects of clomiphene citrate on sperm parameters and fertilisation rate in male NMRI mice

Clomiphene citrate (CC), as a medication in male infertility, improves the sperm parameters in oral consumption but various detrimental side effects have been reported including testicular tumours, gynaecomastia, skin allergic reactions and ocular symptoms. Therefore, this study was designed to evaluate the in vitro effects of CC on sperm parameters and fertilisation rate in IVF protocol. Sperm samples of NMRI adult mice were divided into six groups: group 1 received no treatment (control group), while groups of 2, 3, 4, 5 and 6 (experimental groups) were incubated with the doses of 0.001, 0.01, 0.1, 1 and 10 µg/ml of CC in culture medium respectively. Sperm parameters (viability, morphology and motility), DNA fragmentation levels and fertilisation rate in IVF were evaluated. The results demonstrated that the doses of 0.1 µg/ml (p = .000007 for viability and p = .00006 for fertilisation rate) and 1 µg/ml (p = .032 for viability and p = .005 for fertilisation rate) CC cause a significant improvements; also, the dose of 0.1 µg/ml CC found effective on sperm motility (p = .0003). In the field of IVF, the application of 0.1 and 1 µg/ml of CC in the culture medium may improve the sperm parameters in IVF protocol with no side effects.