• Development and Reproduction
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• v.15 no.2
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• pp.159-166
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• 2011

Simazine (6-chloro-N,N'-diethyl-1,3,5-triazine-2,4-diamine) is a triazine herbicide that has been applied worldwide including Korea for agricultural purposes. Simazine is the second most commonly detected pesticide in surfaceand ground-water in the United States, Europe and Australia. It has been shown that simazine is a potent endocrine disruptor in wildlife and laboratory animals. Although many endocrine disruptors can induce apoptosis in various types of cells, the effects of simazine on apoptosis and on the expression of Bcl-2 family genes are not known. Also it is unknown the effect of simazine on the expression of steroidogenesis-regulating genes in testicular cells. In this study, we investigated the effect of simazine on the expression levels of apoptosis- and steroidogenesis-regulating genes in testicular cells. We found that a low concentration of simazine can alter the mRNA expression levels of steroidogenesis-related genes and Bcl-2 family genes in mouse Sertoli cells and rat Leydig cells. Thus, our results suggest that simazine can disturb normal testicular development and reproductive function by altering the expression of genes that are critical for the regulation of apoptosis and steroidogenesis.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.373-380
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• 2000

Objective: The purpose of this study was to evaluate effects of goat milk and fermented goat milk on reproductive function and stamina of male rodent. Methods: Experiment I: Male ICR mouse was divided into four groups. Group 1 none-treated control; Group 2 received saline; Group 3 received cow milk 10 ml/kg per day for 15 days; Group 4 received goat milk 10 ml/kg per day for 15 days. The cauda epididymal sperm motility and testicular sperm production were investigated. Experiment II: Male SD rat was divided into three groups. Group 1 received saline; Group 2 received goat milk 10 ml/kg per day for 28 days; Group 3 received fermented goat milk 10 ml/kg per day for 28 days. The cauda epididymal sperm motility and testicular sperm production were also investigated. The concentration of testosterone in serum at 1 and 3 weeks after treatment was determined using Immulite 2000 kit. Testes, epididymis, prostate, and seminal vesicle were weighed. Experiment III: Male ICR mouse was divided into four groups. Group 1 none-treated control; Group 2 received saline; Group 3 received goat milk 10 ml/kg per day for 4 weeks; Group 4 received fermented goat milk 10 ml/kg per day for 4 weeks. After treatment, the mouse was forced to swim to test for stamina. Results: In Experiment I, the cauda epididymal sperm motility after in vitro culture for 1 or 3 h was significantly (p<0.05) higher in cow milk and goat milk than in the control and saline. There was no significant difference in the cauda epidymal sperm motility between cow and goat milk. The testicular spermatid number was significantly (p<0.01) higher in goat milk (222.8${\times}10^6$) than in the control (108.6), saline (98.2), and cow milk (118.2). In Experiment II, the cauda epididymal sperm motility after in vitro culture for 1 h was significantly (p<0.05) higher in fermented goat milk than in saline and goat milk. There was no significant difference in the cauda epidymal sperm motility between saline and goat milk but goat milk showed slightly higher sperm motility than saline. After in vitro culture for 3 h, the cauda epididymal sperm motility was significantly (p<0.01) higher in fermented goat milk and goat milk than in saline. The testicular spermatid number was significantly (p<0.05) higher in goat milk than in saline, and significantly (p<0.01) higher in fermented goat milk than in saline. And the serum testosterone levels of rats administered with goat milk or fermented goat milk were increased but were no significant difference among three groups. Also the prostate weight was significantly (p<0.05) increased in the goat and fermented goat milk. In Experiment III, the swimming time in the goat milk and fermented goat milk groups was significantly (p<0.01) longer than in the control and saline. There was no significant difference in the swimming time between goat and fermented goat milk but the fermented goat milk showed slightly longer swimming time than the goat milk. Conclusion: The cauda epididymal sperm motility, the testicular spermatid number and stamina were improved when the mice and rats were drunk with goat milk or fermented goat milk.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.5
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• pp.1180-1186
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• 1999

A wide ranges of chemicals released into the environment have potential to interfere with physiological and development process by disrupting endocrine pathways. Endocrine system embraces a multitude of mechanisms of action, including effect on growth, behavior, reproduction and immune function. These environmental endocrine disruptors are present in environment and pose potential health consequences to human and wildlife. The best known form in endocrine distruptors involves substances which mimic or block the action of natural hormone in the body. Endocrine disruptor have been variously defined as exogenous agents that interfere with the synthesis, secretion, transport, metabolism, binding action or elimination of the natural hormones in the body which are responsible for the maintenance of homeostasis, reproduction developmental and/or behavior. Many compounds polluted into the environment by human activity are capable of disrupting the endocrine system of animals, including fish, wildlife, and humans. Among these chemicals are pesticides, industrial chemicals, and other anthropogenic products. It has been alleged that several adverse effects on human health are linked with exposure to chemicals which are claimed to be endocrine disrupters, that is, increased incidence of testicular, prostate and female breast cancer, time dependent reductions in sperm quality and quantity, increased incidence of cryptorchidism (undescended testicles) and hypospadias(malformation of the penis), altered physical and mental de velopment in children. This observation is currently the only example of chemically mediated endocrine disruption which has resulted in a clear effect at the population level.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.1
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• pp.9-15
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• 2022

The angiotensin-converting enzyme 2 receptor (ACE2) appears to be widely expressed in cells in the testes, predominantly in spermatogonia, Sertoli cells, and Leydig cells, and its co-expression with transmembrane protease serine 2 (TMPRSS2) is essential for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). For this reason, the male reproductive system could be considered a potential target for SARS-CoV-2, as well as a possible reservoir of infection. However, to date, there is very little evidence about the presence of SARS-CoV-2 in semen and testicular samples. The aim of this paper was to review the current evidence regarding the impact of SARS-CoV-2 on male fertility and sexual health, with a particular focus on reproductive hormones, the presence of the virus in seminal fluid and testis, and its impact on fertility parameters. We found very limited evidence reporting the presence of SARS-CoV-2 in semen and testicular samples, and the impact of SARS-CoV-2 on reproductive hormones and fertility parameters is unclear. The quality of the examined studies was poor due to the small sample size and several selection biases, precluding definitive conclusions. Hence, future well-designed prospective studies are needed to assess the real impact of SARS-CoV-2 on male reproductive function.

• Development and Reproduction
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• v.26 no.3
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• pp.99-105
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• 2022

Styrene is the precursor of polystyrene. Human exposure to styrene could occur in occupational and residential settings and via food intake. Styrene is metabolized to styrene-7,8-oxide by cytochrome P450 enzyme. In the present study, we investigated the cytotoxicity mediated by styrene and styrene-7,8-oxide in TM3 testicular Leydig cells in vitro. We first monitored the nuclear fragmentation in Leydig cells after exposure to styrene or styrene-7,8-oxide. Hoechst 33258 cell staining showed that styrene exposure in TM3 Leydig cells did not exhibit nuclear fragmentation at any concentration. In contrast, nuclear fragmentation was seen in styrene-7,8-oxide-exposed cells. These results indicate that cytotoxicity-mediated cell death in Leydig cells is more susceptible to styrene-7,8-oxide than to styrene. Following styrene treatment, procaspase-3 and XIAP protein levels did not show significant changes, and cleaved (active) forms of caspase-3 were not detected. Consistent with the western blot results, the active forms of caspase-3 and XIAP proteins were not prominently altered in the cytoplasm of cells treated with styrene. In contrast to styrene, styrene-7,8-oxide induced cell death in an apoptotic fashion, as seen in caspase-3 activation and increased the expression of XIAP proteins. Taken together, the results obtained in this study demonstrate a fundamental idea that Leydig cells are capable of protecting themselves from cytotoxicity-mediated apoptosis as a result of styrene exposure in vitro. It remains unclear whether the steroid-producing function, i.e., steroidogenesis, of Leydig cells is also unaffected by exposure to styrene. Therefore, further studies are needed to elucidate the endocrine disrupting potential of styrene in Leydig cells.

• Korean Journal of Agricultural Science
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• v.48 no.4
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• pp.935-944
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• 2021

It has been suggested that bisphenol A (BPA), a known endocrine disruptor, interferes with the endocrine system, causing reproductive dysfunction. Recently, BPA has been found in waste water due to incomplete sewage purification, possibly threatening health through its ingestion via tap water. In this study, young male mice (6 - 7 weeks old) were administered water containing BPA (50 mg·kg^-1) for four weeks, while control mice consumed water without BPA. Serum, epididymal spermatozoa and testicular sections were assessed after sacrificing the mice on day 28. No significant differences were obtained between the groups in the body, testis and seminal vesicle weights. However, the epididymal sperm motility and count levels were significantly reduced in BPA-fed mice. Significantly higher hepatotoxicity levels were also observed in mice ingesting BPA as compared to the control mice. The level of serum testosterone was reduced, and testicular sections revealed incomplete and irregular spermatogenesis in BPA-ingested mice. The sperm proteasomal-proteolytic activity level has been implicated in sperm function and is measured in motile spermatozoa using fluorometric substrates. High ubiquitin C-terminal hydrolase activity levels were observed in the control mice without BPA. During a mating trial, a low pregnancy rate (71.4%) was observed in females mated with males who had consumed BPA (100% in the control mice). Overall, BPA adversely affected spermatogenesis and quality, as indicated by decreased sperm motility, concentration and serum testosterone levels, resulting in reduced fertility competence.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.4
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• pp.291-299
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• 2023

Background: Leydig cells, crucial for testosterone production, express ion channels like ANO1 that influence hormone secretion. This study investigates the expression and role of the Tandem of P domains in a weak inward rectifying K⁺ channel-related Acid-Sensitive K⁺-1 (TASK-1) channel in these cells, exploring its impact on testicular function and steroidogenesis. Methods: TASK-1 expression in Leydig cells was confirmed using immunostaining, while RT-PCR and Western Blot (WB) validated its expression in the TM3 Leydig cell line. The effect of a TASK-1 channel blocker on cell viability was assessed through live/dead staining and MTT assays. Additionally, the blocker's effect on testosterone secretion was evaluated by measuring testosterone levels. Results: Immunohistochemical analysis revealed a predominant presence of TASK-1, along with c-Kit and ANO-1, in Leydig cells adjacent to seminiferous tubules and also in Sertoli and spermatogenic cells. Expression levels of TASK-1 mRNA and protein were significantly higher in TM3 Leydig cells compared to TM4 Sertoli cells. In addition, blocking TASK-1 in TM3 cells with ML365 induced cell death but did not affect LH-induced testosterone secretion. Conclusions: These findings suggest that TASK-1 in Leydig cells is crucial for their viability and proliferation, highlighting its potential importance in testicular physiology.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.1
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• pp.48-56
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• 2024

Objective: This study investigated the relationship of anthropometric and metabolic risk factors with seminal and sex steroidal hormone parameters in a screened population of healthy males. Methods: The participants were healthy young men without chronic or congenital diseases. The body composition parameters that we investigated were measured weight, height, and waist circumference (WC), as well as bioelectrical impedance analysis. Semen samples were analyzed for semen volume, sperm concentration, sperm motility and morphology, seminal pH, and liquefaction time. Biochemistry analysis, including glucose and lipid metabolism parameters, was conducted on fasting blood samples. Testicular volume was calculated separately for each testis using ultrasonography. Results: Body mass index exhibited an inverse association with total sperm count. WC showed negative correlations with numerous seminal parameters, including sperm concentration, total sperm count, sperm morphology, and follicle-stimulating hormone levels. The basal metabolic rate was associated with seminal pH, liquefaction time, and sperm motility. WC, fat mass percentage, and triglyceride levels exhibited negative correlations with sex hormone binding globulin. The measures of glucose metabolism were associated with a greater number of seminal parameters than the measures of cholesterol metabolism. C-reactive protein levels were inversely associated with sperm concentration and total sperm count. Conclusion: Anthropometric and metabolic risk factors were found to predict semen quality and alterations in sex steroidal hormone levels.

• Anatomy and Cell Biology
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• v.57 no.1
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• pp.119-128
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• 2024

Glucocorticoids play a physiologic role in the adult male reproductive functions, modulating gonadal steroid synthesis and spermatogenesis, through the glucocorticoid receptor (GR). The expression of GR has been described in several key testicular cell types, including somatic cells and early germ cell populations. Nothing is known on GR in human spermatozoa. Herein, we explored the GR expression and its possible role in normal and testicular varicocele semen samples from volunteer donors. After semen parameter evaluation by macro- and microscopic analysis, samples were centrifuged; then spermatozoa and culture media were recovered for further investigations. By western blotting and immunofluorescence analyses we evidenced for the first time in spermatozoa the presence of GR-D3 isoform which was reduced in sperm from varicocele patients. By treating sperm with the synthetic glucocorticoid dexamethasone (DEXA), we found that survival, motility, capacitation, and acrosome reaction were increased in both healthy and varicocele samples. GR involvement in mediating DEXA effects, was confirmed by using the GR inhibitor mifepristone (M2F). Worthy, we also discovered that sperm secretes different cortisol amounts depending on its physio-pathological status, suggesting a defence mechanism to escape the immune system attach in the female genital tract thus maintaining the immune-privilege as in the testis. Collectively, our data suggests a role for glucocorticoids in determining semen quality and function, as well as in participating on sperm immune defensive mechanisms. The novelty of this study may be beneficial and needs to take into account in artificial insemination/drug discovery aimed to enhancing sperm quality.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.189-197
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• 2018

Objective: Hemicastration is a unilateral orchiectomy to remove an injured testis, which can induce hormonal changes and compensatory hypertrophy of the remaining testis, and may influence spermatogenesis. However, the underlying molecular mechanisms are poorly understood. Here, we investigated the impact of hemicastration on remaining testicular function. Methods: Prepubertal mice (age 24 days) were hemicastrated, and their growth was monitored until they reached physical maturity (age 72 days). Subsequently, we determined testis DNA methylation patterns using reduced representation bisulfite sequencing of normal and hemicastrated mice. Moreover, we profiled the testicular gene expression patterns by RNA sequencing (RNA-seq) to examine whether methylation changes affected gene expression in hemicastrated mice. Results: Hemicastration did not significantly affect growth or testosterone (p>0.05) compared with control. The genome-wide DNA methylation pattern of remaining testis suggested that substantial genes harbored differentially methylated regions (1,139) in gene bodies, which were enriched in process of protein binding and cell adhesion. Moreover, RNA-seq results indicated that 46 differentially expressed genes (DEGs) involved in meiotic cell cycle, synaptonemal complex assembly and spermatogenesis were upregulated in the hemicastration group, while 197 DEGs were downregulated, which were related to arachidonic acid metabolism. Integrative analysis revealed that proteasome 26S subunit ATPase 3 interacting protein gene, which encodes a protein crucial for homologous recombination in spermatocytes, exhibited promoter hypomethylation and higher expression level in hemicastrated mice. Conclusion: Global profiling of DNA methylation and gene expression demonstrated that hemicastration-induced compensatory response maintained normal growth and testicular morphological structure in mice.