Spermatogenic Function Research Articles

Protective effects of HSP70, HO-1 and HSP27 on spermatogenic function in rats with varicocele

Objective To investigate the protective effects of HSP70、HO-1 and HSP27 on spermatogenic function in rats with varicocele. Methods Forty adult male SD rats were randomly divided into 4 groups with 10 rats in each group: sham group (group A), sham + quercetin group (group B), VC group (group C) and VC+ quercetin group (group D). An experimental varicocele model was successfully created in group C and D and quercetin(100 mg/kg) was injected intraperitoneally. HE staining was used to observe the pathological changes of testis. Malondialdehyde (MDA) and superoxide dismutase (SOD) content were determined. Cell apoptosis index was detected by dUTP nick end labeling. The expression of HSP70 in testis tissue was detected by immunohistochemistry. The mRNA expression of HSP27 and HO-1 in testis tissue was measured by RT-PCR. Results Testicular structure and spermatogenic function were seriously affected, and histological damage was significantly increased caused by VC in group D. AI was (5.26±0.45)% , (5.83±0.52)% , (14.64±0.98)% , (25.37±1.46)%, respectively.The SOD activities were(0.96±0.06), (0.88±0.05), (0.45±0.04), (0.23±0.03) U/mg protein, MDA content was(2.24±0.35), (2.69±0.32), (4.78±0.41), (7.37±0.68) nmol/g protein.The expression of HSP70 in group C was significantly higher than that in group A and B, and the expression of HSP70 in group D was significantly lower than that of group C. Compared with group A and B, the mRNA expression level of HSP27 and HO-1 was significantly increased in group C. Compared with group C, the expression level of HSP27 and HO-1 in group D was significantly decreased.The comparison between B、C and D groups was statistically significant (P<0.05). Conclusions HSP70、HO-1 and HSP27 reduces the level of oxidative stress and inhibits the apoptosis.Thus, it plays a protective role on the spermatogenic function in rats with varicocele. Key words: Varicocele; Disease Models, Animal; HSC70 Heat-Shock Proteins; Heat-Shock Proteins; Rats

SPERM QUALITY IN RATS PREDISPOSED TO THE MANIFESTATION OF CATATONIC REACTIONS

Catatonia is a psychopathological syndrome displayed as a motor disorder. Catatonia is a sign of many menthal disorders, particularly schizophrenia and depression, with a wide disrtibution in the human population. The GC (“genetic” and “catatonia”) rat strain was obtained from the Wistar rat strain by a long selection (78 generations) for the catatonic type of reaction and is a model of schizophrenic and depressive disorders in humans. It is known that selection for behavior including catatonic reactions results in neuroendocrine, reproductive and morphological changes in animals. However, the influence of selection for a catatonic reaction on the spermatogenic function of testes had not been studied. The aim of this study was to conduct a comparative investigation of sperm quality in rats of the GC and the Wistar strain. The epididymal sperm parameters (sperm count, sperm motility, sperm morphology) were measured, and body, testes and epididymal weight were determined at puberty (50 day of life) and at adulthood (90 day of life). The litter size of the GC and Wistar rats was determined. It was found that adult GC rats had a lower sperm count, sperm motility, testis weight, epydidymal weight and litter size compared to adult Wistar rats. However, at puberty, GC rats had a higher sperm count than the Wistar strain. Interstrain differences in sperm morphology were not found. It has been assumed that the changes of spermatogenic parameters in response to selection for catatonia are caused by changing the ontogenic pattern of testosterone secretion. In conclusion, the hereditary predisposition to catatonic reaction is associated with impaired sperm parameters in adult rats that reduces their chance to reproduction. The GC rat strain can be a perspective model for investigation of the relationship between the hereditary predisposition to catatonia and spermatogenesis

CTRP3 attenuates high-fat diet-induced male reproductive dysfunction in mice.

Recent studies have suggested a role for abdominal obesity in male infertility. Previous studies have found that cell apoptosis exerts an important role in obesity-related male infertility. C1q/TNF-related protein 3 (CTRP3), a paralog of adiponectin, has been proposed to exert anti-apoptotic effects and to attenuate diabetes-related cardiac injuries. However, the role of CTRP3 in high-fat diet (HFD)-induced spermatogenic impairment remains unclear. In the present study, we fed male mice an HFD for 24 weeks to induce obesity. The expression of CTRP3 was decreased by HFD feeding. Supplementation with the recombinant human globular domain of CTRP3 (0.25 μg/g/day) for 4 weeks beginning at 20 weeks of the HFD improved spermatogenic function in the HFD-fed mice, which were characterized by improved testis morphology, increased testis weight/body weight ratio, and increased sperm count, sperm viability, and sperm motility. We also found that CTRP3 infusion resulted in the attenuation of endoplasmic reticulum (ER) stress and the activation of silence information regulator 1 (SIRT1) in the testes of obese mice. Our in vitro study also suggested that CTRP3 attenuated the palmitic acid (PA)-induced reductions in sperm viability and motility via the inhibition of ER stress. Moreover, germ cell-specific Sirtuin1 knockout abolished the protective effects of CTRP3 in vivo and in vitro. In vitro studies of human sperm showed that the protective effects of CTRP3 on sperm viability and motility were abrogated by a specific inhibitor of SIRT1. Thus, our results demonstrated that CTRP3 expression protected against HFD-induced spermatogenic deficiency through the SIRT1/ER stress pathway.

Taurine enhances spermatogenic function and antioxidant defense mechanisms in testes and epididymis of L-NAME-induced hypertensive rats

The beneficial health effects of taurine on hypertension have been demonstrated previously in both experimental and epidemiological studies. However, the role of taurine in reproductive dysfunction associated with hypertension has not been investigated. The present study evaluated the therapeutic efficacy of taurine on reproductive deficits in N-nitro-l-arginine methyl ester (L-NAME)-induced hypertensive rats. Sixty male Wistar rats were randomly assigned into six groups namely control, taurine alone, L-NAME alone (40mg/kg) or L-NAME treated with either taurine (100 and 200mg/kg) or reference drug atenolol (10mg/kg) for 28 consecutive days. Results indicated that taurine treatment significantly abrogated L-NAME-induced increase in systolic, diastolic and mean arterial pressures when compared with hypertensive control. Administration of taurine markedly increased antioxidant enzymes activities and glutathione level whereas it suppressed the increase in biomarkers of oxidative stress in the testes and epididymis of L-NAME-induced hypertensive rats. Moreover, taurine significantly reversed hypertension mediated decreases in circulatory concentrations of luteinizing hormone, follicle-stimulating hormone and testosterone whereas it increased testicular sperm number, epididymal sperm number and sperm progressive motility in the hypertensive rats. Furthermore, taurine abrogated the suppression of marker enzymes of testicular function namely acid phosphatase, alkaline phosphatase and lactate dehydrogenase and preserved the histo-architectures of the testes and epididymis in L-NAME-induced hypertensive rats. Taken together, the findings from this study highlight the beneficial role of taurine in reproductive system of L-NAME-induced male hypertensive rats. Taurine supplementation may be a good clinical approach to prevent reproductive deficits in male hypertensive patients.

Bovine thyroglobulin gene polymorphisms and their association with sexual precocity in Guzerat bulls.

Puberty is a stage of sexual development determined by the interaction of environmental factors and genetic mechanisms. Among them, thyroid function plays a key role in sexual development and spermatogenic function and is under the control of several genes, including the well-described thyroglobulin gene (TG). Previous reports have shown genetic association between thyroid function and selected single nucleotide polymorphisms (SNPs) in taurine cattle. Therefore, the identification of genetic mechanisms involved in the regulation of this trait can assist with the selection for early pubertal bulls, thus improving genetic progress in livestock breeding. The aim of this study was to validate the association between TG SNPs and age at puberty in zebuine bulls. Three SNPs (rs110406764, rs109662686, rs109057985) were genotyped in 159 Guzerat animals using SEQUENOM technology. Results showed a significant association (p<.05) between the studied SNPs and puberty age, in agreement with our previous reports in a taurine breed. Interestingly, allele frequencies were different from those already reported, being GAT the most favourable allele for age at puberty in Guzerat (94.4days lower). Overall, our findings corroborate previous reports and reinforce the importance of genetic influence in the regulation of sexual development and puberty through a thyroid pathway in zebuine cattle.

Spermatogenic failure and the Y chromosome.

The Y chromosome harbors a number of genes essential for testis development and function. Its highly repetitive structure predisposes this chromosome to deletion/duplication events and is responsible for Y-linked copy-number variations(CNVs) with clinical relevance. The AZF deletions remove genes with predicted spermatogenic function en block and are the most frequent known molecular causes of impaired spermatogenesis (5-10% of azoospermic and 2-5% of severe oligozoospermic men). Testing for this deletion has both diagnostic and prognostic value for testicular sperm retrieval in azoospermic men. The most dynamic region on the Yq is the AZFc region, presenting numerous NAHR hotspots leading to partial losses or gains of the AZFc genes. The gr/gr deletion (a partial AZFc deletion) negatively affects spermatogenic efficiency and it is a validated, population-dependent risk factor for oligozoospermia. In certain populations, the Y background may play a role in the phenotypic expression of partial AZFc rearrangements and similarly it may affect the predisposition to specific deletions/duplication events. Also, the Yp contains a gene array, TSPY1, with potential effect on germ cell proliferation. Despite intensive investigations during the last 20years on the role of this sex chromosome in spermatogenesis, a number of clinical and basic questions remain to be answered. This review is aimed at providing an overview of the role of Y chromosome-linked genes, CNVs, and Y background in spermatogenesis.

Diffusion-Weighted and Magnetization Transfer Imaging in Testicular Spermatogenic Function Evaluation: Preliminary Results.

To assess the use of functional magnetic resonance imaging (MRI), including diffusion-weighted MRI (DWI) and magnetization transfer MRI (MTI) in evaluating male infertility. Sixteen men with testicular spermatogenesis hypofunction confirmed by percutaneous testis biopsy and 31 volunteers (control group B, age range: 20-40 years) with normal semen analysis including younger (By, n = 15, age range: 20-30 years) and older (Bo, n = 16, age range: 31-40 years) men underwent pelvic 3T MRI, including DWI and MTI. Apparent diffusion coefficient (ADC) and magnetization transfer ratio (MTR) were compared. The ADCs in 32 testes of 16 patients (0.497 ± 0.037 × 10-3 mm2 /s) were significantly (P < 0.001) higher than that of control group B (0.460 ± 0.031 × 10-3 mm2 /s), group By (0.453 ± 0.018 × 10-3 mm2 /s), and group Bo (0.461 ± 0.034 × 10-3 mm2 /s), whereas the MTRs were significantly lower than that of group B (16.14 ± 4.20), group By (17.88 ± 2.00), and group Bo (15.09 ± 4.28) (P < 0.001). Functional MRI, including DWI and MTI, appears promising for evaluating male infertility with higher ADC and lower MTR in testicular spermatogenesis hypofunction. 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2018;47:186-190.

Mechanisms of Heshouwuyin in regulating apoptosis of testicular cells in aging rats through mitochondrial pathway.

BackgroundPolygonum multiflorum has important effects on anti-aging and immunity enhancement. Many traditional Chinese medicine preparations based on Polygonum multiflorum are widely used for the clinical prevention and treatment of aging. However the mechanisms of these herb mixtures are often unknown. This study investigates the effect of Heshouwuyin, a Chinese herbal compound for invigorating the kidney, on the regulation of testicular cells apoptosis in aging rats.MethodsIn this study, 18-month-old Wistar rats served as a model of natural aging and 12-month-old rats served as a young control group. Heshouwuyin group 1 and group 2 were comprised 18-month-old rats given Heshouwuyin intragastrically for 60 days and 30 days respectively. Then testes of the young control group were isolated in the age of 12 months, the other three groups were in the age of 18 months.ResultsTUNEL assay showed that the rate of testicular cell apoptosis was obviously higher and Flow cytometry analysis showed that the rate of cell proliferation was significantly lower in the natural aging group than in the young control group and that intervention with Heshouwuyin could reverse this phenomenon. Therefore, we further applied microarray analysis to screen out differentially expressed genes regulated by Heshouwuyin and related to cell apoptosis. The expression of these genes was observed by quantitative fluorescence PCR, immunofluorescence staining, and western blot. The results showed that the expression of 14-3-3σ was significantly lower and that the expression of DR6, BAX, caspase-3 and Cytc were significantly higher in the natural aging group than in the young control group, but intervention with Heshouwuyin significantly reversed this phenomenon. Moreover, the curative efficacy of Heshouwuyin after 60 days was better than that of Heshouwuyin after 30 days.ConclusionOur study suggests that Heshouwuyin has anti-aging effects on the testis by means of inhibiting the occurrence of apoptosis in spermatogenic cells, thus improving the spermatogenic function of the testis. This is mainly achieved by regulating the expression of key genes in the mitochondrial apoptosis pathway.

Mapping the testicular interstitial fluid proteome from normal rats.

Communication between the testicular somatic (Sertoli, Leydig, peritubular myoid, macrophage) and germ cell types is essential for sperm production (spermatogenesis), but the communicating factors are poorly understood. We reasoned that identification of proteins in the testicular interstitial fluid (TIF) that bathes these cells could provide a new means to explore spermatogenic function. The aim of this study was to map the proteome of TIF from normal adult rats. Low-abundance proteins in TIF were enriched using ProteoMiner beads and identified by MALDI-MS/MS, recognizing 276 proteins. Comparison with proteomic and genomic databases showed these proteins originated from germ cells, somatic cells (Sertoli, peritubular myoid, Leydig), and blood plasma. In silico analysis revealed homologues of >80% TIF proteins in the human plasma proteome, suggesting ready exchange between these fluids. Only 36% of TIF proteins were common with seminiferous tubule fluid that transports mature spermatids to the epididymis, indicating these two fluids are quite different. This TIF proteome provides an important new resource for the study of intercellular communication in the testis.

Effects of 4-nonylphenol on spermatogenesis and induction of testicular apoptosis through oxidative stress-related pathways.

This study tested the hypothesis that prepubertal exposure to 4-nonylphenol (NP) affects reproductive function in male rats. Twenty-four rats at five-weeks-old were randomly divided into four groups and treated with NP at varying concentrations (0, 5, 20, and 60mg/kg/2d) for thirty days by intra-peritoneal injection. 60mg/kg NP induced spermatogenic degeneration and pronounced deficits in epididymal sperm count, motility and function, whereas potentially stimulatory effects were observed at 5 NPmg/kg. Moreover, 60mg/kg NP resulted in a significant reduction in fructose, FSH and LH; induced apoptosis related to oxidative stress; inhibited mRNA and protein levels of Bcl-2 and PCNA; as well as the additional up-regulation of p53, Bax, Apaf-1, cytochrome c, cleaved-caspase-3, Fas and FasL expression. Our data suggest potentially hormetic effects of NP on spermatogenic function. High-dose NP impairs testicular development and function by reducing cell proliferation and inducing apoptosis involving oxidative stress-related p53-Bcl-2/Bax and -Fas/FasL pathways.

A trial of semen collection by transrectal electroejaculation method from Amur leopard cat (&lt;i&gt;Prionailurus bengalensis euptilurus&lt;/i&gt;)

We collected semen from a male Amur leopard cat using the transrectal electroejaculation method and investigated the semen qualities for about four years. In addition, the influence of the season on the spermatogenic function of the Amur leopard cat was investigated with regard to the semen qualities, testicular volume and serum testosterone level. As a result, we could collect semen with good sperm qualities that would be useable for artificial insemination. Some seasonality was noted in the testicular volume and serum testosterone level. We clarified that the semen qualities were favorable before and during the female breeding season compared with those after the breeding season.

Silencing of X-Linked MicroRNAs by Meiotic Sex Chromosome Inactivation.

During the pachytene stage of meiosis in male mammals, the X and Y chromosomes are transcriptionally silenced by Meiotic Sex Chromosome Inactivation (MSCI). MSCI is conserved in therian mammals and is essential for normal male fertility. Transcriptomics approaches have demonstrated that in mice, most or all protein-coding genes on the X chromosome are subject to MSCI. However, it is unclear whether X-linked non-coding RNAs behave in a similar manner. The X chromosome is enriched in microRNA (miRNA) genes, with many exhibiting testis-biased expression. Importantly, high expression levels of X-linked miRNAs (X-miRNAs) have been reported in pachytene spermatocytes, indicating that these genes may escape MSCI, and perhaps play a role in the XY-silencing process. Here we use RNA FISH to examine X-miRNA expression in the male germ line. We find that, like protein-coding X-genes, X-miRNAs are expressed prior to prophase I and are thereafter silenced during pachynema. X-miRNA silencing does not occur in mouse models with defective MSCI. Furthermore, X-miRNAs are expressed at pachynema when present as autosomally integrated transgenes. Thus, we conclude that silencing of X-miRNAs during pachynema in wild type males is MSCI-dependent. Importantly, misexpression of X-miRNAs during pachynema causes spermatogenic defects. We propose that MSCI represents a chromosomal mechanism by which X-miRNAs, and other potential X-encoded repressors, can be silenced, thereby regulating genes with critical late spermatogenic functions.

Differences and similarities between extremely severe oligozoospermia and cryptozoospermia in intracytoplasmic sperm injection.

Patients with extremely severe oligozoospermia (ESO) and cryptozoospermia (CO) are suitable using intracytoplasmic sperm injection (ICSI) as an infertility treatment. However, some andrologists are confused to distinguish ESO and CO in clinic diagnose. This study was designed for the first time to evaluate and compare patients with ESO and CO to determine whether these are useful clinical distinctions. A total of 270 infertile men in our center were classified into four groups as Group nonobstruction azoospermia (NOA, n = 44), Group ESO (n = 78), Group CO (n = 40), and Group obstruction azoospermia (OA, n = 108). Comparisons of the volume of bilateral testes, the level of follicle stimulating hormone (FSH) and inhibin B were obtained in four groups. Then comparisons of fertilization rates, cleavage rate, and excellent embryos rate were obtained when couples performed ICSI. All indexes (volume of bilateral testis, level of FSH and inhibin B) in Groups ESO and CO were no difference, while Groups OA versus NOA, OA versus ESO, and OA versus CO were significant differences (P < 0.05). The rates of fertilization were no differences in Groups ESO and CO while Groups OA versus ESO, OA versus CO were significant differences (P < 0.05). Therefore, the spermatogenic functions in patients with CO and ESO were similar, better than NOA but worse than OA. However, it would be helpful to evaluate their spermatogenesis using testicular biopsies, especially accompanied azoospermia in clinical practice.

Beyond Testis Size: Links between Spermatogenesis and Sperm Traits in a Seasonal Breeding Mammal.

Spermatogenesis is a costly process that is expected to be under selection to maximise sperm quantity and quality. Testis size is often regarded as a proxy measure of sperm investment, implicitly overlooking the quantitative assessment of spermatogenesis. An enhanced understanding of testicular function, beyond testis size, may reveal further sexual traits involved in sperm quantity and quality. Here, we first estimated the inter-male variation in testicular function and sperm traits in red deer across the breeding and non-breeding seasons. Then, we analysed the relationships between the testis mass, eight parameters of spermatogenic function, and seven parameters of sperm quality. Our findings revealed that the Sertoli cell number and function parameters vary greatly between red deer males, and that spermatogenic activity co-varies with testis mass and sperm quality across the breeding and non-breeding seasons. For the first time in a seasonal breeder, we found that not only is the Sertoli cell number important in determining testis mass (r = 0.619, p = 0.007 and r = 0.248, p = 0.047 for the Sertoli cell number assessed by histology and cytology, respectively), but also sperm function (r = 0.703, p = 0.002 and r = 0.328, p = 0.012 for the Sertoli cell number assessed by histology and cytology, respectively). Testicular histology also revealed that a high Sertoli cell number per tubular cross-section is associated with high sperm production (r = 0.600, p = 0.009). Sperm production and function were also positively correlated (r = 0.384, p = 0.004), suggesting that these traits co-vary to maximise sperm fertilisation ability in red deer. In conclusion, our findings contribute to the understanding of the dynamics of spermatogenesis, and reveal new insights into the role of testicular function and the Sertoli cell number on testis size and sperm quality in red deer.

Protective effects of metformin on reproductive function in obese male rats induced by high-fat diet.

The study aims to elucidate the changes in testicular spermatogenic function in high-fat diet (HFD)-induced obese rats and to evaluate the protective effects of metformin intervention. Male Sprague-Dawley rats (n = 18) were randomly divided into a control group (standard diet), an HFD group, and a metformin group (HFD + metformin at 100 mg/kg, once daily by oral gavage). After 8 weeks, rats were euthanized, and the weights of body and testes were measured. Testis and epididymis were dissected and hematoxylin-eosin-stained for histopathological examination and semen parameter analysis. Blood samples were collected for assessment of sex hormones and metabolic parameters (serum glucose, insulin, and leptin). Spermatogenic cell apoptosis was accessed by TUNEL. Compared with the control group, the final body weight and weight gain were significantly higher in HFD rats, while the testicle weight and coefficients were lower. In HFD rats, metformin treatment induced weight loss and increased testicle weight (P < 0.05). In HFD rats, obvious pathological changes in the testicular tissue were characterized by small, atrophic, and distorted seminiferous tubules and destroyed basement membrane. Metformin treatment protected against the HFD-induced decrease in the number of spermatogonia, Sertoli cells, and Leydig cells (P < 0.05); ameliorated the HFD-induced increases in serum glucose, insulin, leptin, and estrogen; and decreased serum testosterone (P < 0.05) and reduced the rate of testicular cell apoptosis in obese male rats. Finally, metformin significantly improved semen parameters (including concentration, viability, motility, and normal morphology) in HFD rats (P < 0.05). HFD-induced obesity in rats results in detrimental effects on spermatogenesis, semen quality, endogenous hormones, and testicular cell apoptosis. Metformin intervention improved the semen parameters, possibly due to its effects on weight loss, increased testicular weight, reduced testicular cell apoptosis, and resulted in restoration of hormonal homeostasis and correction of metabolic disorder.

Functional significance of the sex chromosomes during spermatogenesis.

Mammalian sex chromosomes arose from an ordinary pair of autosomes. Over hundreds of millions of years, they have evolved into highly divergent X and Y chromosomes and have become increasingly specialized for male reproduction. Both sex chromosomes have acquired and amplified testis-specific genes, suggestive of roles in spermatogenesis. To understand how the sex chromosome genes participate in the regulation of spermatogenesis, we review genes, including single-copy, multi-copy, and ampliconic genes, whose spermatogenic functions have been demonstrated in mouse genetic studies. Sex chromosomes are subject to chromosome-wide transcriptional silencing in meiotic and postmeiotic stages of spermatogenesis. We also discuss particular sex-linked genes that escape postmeiotic silencing and their evolutionary implications. The unique gene contents and genomic structures of the sex chromosomes reflect their strategies to express genes at various stages of spermatogenesis and reveal the driving forces that shape their evolution.Free Chinese abstract: A Chinese translation of this abstract is freely available at http://www.reproduction-online.org/content/149/6/R265/suppl/DC1.Free Japanese abstract: A Japanese translation of this abstract is freely available at http://www.reproduction-online.org/content/149/6/R265/suppl/DC2.

Twenty-four-hour monitoring of scrotal temperature in obese men and men with a varicocele as a mirror of spermatogenic function.

How do day and night scrotal temperatures, spermatogenesis parameters, sex hormones and intratesticular perfusion in obese men and men with a varicocele compare with healthy controls? Compared with healthy controls, 24-h monitoring of scrotal temperature in men with a varicocele and obese men showed higher temperatures and this condition was related to a significant alteration of spermatogenesis and stasis of testicular perfusion. Several studies have shown that increased scrotal temperature has dramatic effects on spermatogenesis. Scrotal hyperthermia by exposure to sauna is able to induce a significant alteration of sperm production. In a case-control study, data were collected over a period of 2 years from 60 subjects with risk factors for testicular heating and 20 healthy subjects who consecutively attended an andrology unit as participants in an infertility prevention program. Forty subjects with a left varicocele, 20 obese men and 20 healthy subjects who served as controls, were evaluated for testicular volumes, sex hormones, sperm parameters, sperm aneuploidies, mean transit time (MTT) of intratesticular blood and 24-h scrotal temperature monitoring by a cutaneous thermochip. Subjects with a varicocele were further subgrouped on the basis of normo or oligozoospermia (VN and VO). Student's t-test was used for statistical analysis. We found a significant increase in 24-h mean scrotal temperature in obese men and men with a varicocele compared with controls (both P < 0.01). This increase in scrotal temperature was associated with impaired sperm parameters and higher FSH plasma levels compared with controls. Dynamic evaluation of scrotal temperatures showed wide fluctuations in controls, but little variation in obese men and men with a varicocele. Men with VO had left and right increase in scrotal temperatures (the right was increased also versus VN, P < 0.01) (both P < 0.001). Men with VN showed a left scrotal temperature higher than controls (P < 0.01) and a right scrotal temperature no different from controls (34.92 ± 0.53 and 34.66 ± 0.65, respectively). Mean MTT values recorded in men with VO were significantly higher than men with VN and obese men (both P < 0.001). Different lifestyle, diet, occupation, stress level and environmental temperatures due to seasonal conditions are major limitations of this study. Our data suggested for the first time that dynamic evaluation of scrotal temperatures seems to reflect alterations of testicular function and perfusion in obese men and men with a varicocele. In these clinical conditions, spermatogenic impairment and scrotal heating seem to be related to different mechanisms. The dynamic evaluation of scrotal temperature in subjects with risk factors for testicular heating could allow the identification of subjects needing treatment or a change in lifestyle. No external funding was sought for this study, and the authors have no conflict of interest to declare.

Effects of malathion on testicular spermatogenic function in rats

To investigate the effects of malathion on the testicular spermatogenic function of male rats and its working mechanism. Forty specific pathogen-free male Wistar rats were randomly and equally divided into four groups: three exposure groups and a control group. Malathion was administered orally to male rats in the exposure groups at 33.75, 54, and 108 mg/kg (1/32 LD₅₀, 1/20 LD₅₀, and 1/10 LD₅₀) for 60 days. Rats in the control group received an equal volume of water. The body weights of rats were measured after exposure. The organ weights and coefficients of the testes and epididymes were determined as soon as rats were sacrificed. The sperm motility, counts, and malformation rates were measured in the left epididymis. Histopathological changes, cell apoptosis, and the expression levels of Bcl-2/Bax in the testes of rats were observed using HE staining, terminal deoxynucleotidyl transferase-mediated dUPT-biotin nick end labeling, and immunohistochemistry SABC method. The body weights and the testis weights in the exposure groups were significantly lower than those in the control group (P < 0.01). The exposure groups had significantly lower sperm motility and significantly higher sperm malformation rates than the control group (P < 0.01). The sperm counts were significantly lower in the exposure groups than in the control group (P<0.01). The sperm counts and motility were negatively correlated with exposure dose (r = -0.81, P < 0.01; r = -0.51, P < 0.01), while the sperm malformation rate was positively correlated with exposure dose (r = 0.85, P 0.01). The exposure groups had significantly higher spermatogenic cell apoptosis rates than the control group (P<0.01). The expression level of Bax was significantly higher in the exposure groups than in the control group (P<0.01), while the expression level of Bcl-2 was significantly lower in the exposure groups than in the control group (P < 0.01). Histopathological examination of the testes showed degenerative changes in the seminiferous tubules at various doses along with the increase in malathion exposure dose. Malathion affects the testicular spermatogenic function of male rats and its working mechanism may involve cell apoptosis induced by down-regulation of Bcl-2 and up-regulation of Bax.

Role of Inhibitors of Apoptosis Proteins in Testicular Function and Male Fertility: Effects of Polydeoxyribonucleotide Administration in Experimental Varicocele.

Neuronal apoptosis inhibitory protein (NAIP) and survivin might play an important role in testicular function. We investigated the effect of PDRN, an agonist of adenosine A2A receptor, on testicular NAIP and survivin expression in an experimental model of varicocele. After the creation of experimental varicocele (28 days), adolescent male Sprague-Dawley rats were randomized to one of the following treatments lasting 21 days: vehicle, PDRN (8 mg/kg i.p., daily), PDRN + 3,7-dimethyl-propargylxanthine (DMPX, a specific adenosine A2A-receptor antagonist, 0.1 mg/kg i.p., daily), varicocelectomy, and varicocelectomy + PDRN (8 mg/kg i.p., daily). Sham-operated animals were used as controls. Animals were then euthanized and testis expression of NAIP and survivin was evaluated through qRT-PCR, western blot, and immunohistochemical analysis. Spermatogenetic activity was also assessed. NAIP and survivin expressions were significantly reduced following varicocele induction when compared to sham animals whereas PDRN-treated rats showed an increase in NAIP and survivin levels. Immunohistochemistry revealed an enhanced expression of NAIP and survivin with a characteristic pattern of cellular localization following PDRN treatment. Moreover, administration of PDRN significantly restored spermatogenic function in varicocele rats. PDRN may represent a rational therapeutic option for accelerating recovery from depressed testicular function through a strategic modulation of apoptosis in experimental varicocele.

Montelukast prevents testes against ischemia-reperfusion injury through suppression of iNOS expression.

To elucidate the mechanism of a possible protective effect of montelukast against testicular ischemia/reperfusion (I/R) injury. Fifty-one adult male Wistar-Albino rats were randomly assigned into 6 groups; sham + saline (S), sham + montelukast (M), I/R + S, I/R + S 30', I/R + M and I/R + M 30'. Saline or montelukast (10 mg/kg) was intraperitoneally administered 30 minutes prior to (S 30', M 30') and during detorsion (I/R + S, I/R + M) in the I/R groups. The I/R groups underwent 2 hours of ischemia followed by 4 hours (early-term) of reperfusion in unilateral testes. Half of the rats underwent 24 hours (late-term) of reperfusion to investigate long-term effects. Testicular tissue samples were examined for biochemical and histopathological parameters. Germ cell apoptosis was evaluated using apoptosis-activating factor 1 (Apaf-1). Inducible nitric oxide synthase (iNOS) activity was analyzed in late-term reperfusion groups. Spermatogenic functions were assessed for each testis based on the Johnsen criteria. Unilateral I/R caused a significant increase in serum TNF-α levels in the early-term group compared to the sham groups. Malondialdehyde levels and myeloperoxidase activity were found to be elevated in the I/R groups and accompanied with a significant decrease in glutathione levels when compared to the sham groups. I/R significantly increased iNOS activity and germ cell apoptosis compared to the sham groups. Montelukast treatment significantly reversed all of these parameters and achieved comparable results with the sham groups. Finally, spermatogenic indices were similar for the bilateral testes between all groups. Montelukast exerts protective effects against testicular I/R injury by inhibiting neutrophil activity, reversing the oxidative stress markers, decreasing iNOS activity and attenuating apoptosis.