• Clinical and Experimental Reproductive Medicine
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• v.47 no.2
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• pp.101-107
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• 2020

Objective: The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. Methods: Thirty-two adult male mice were divided equally into four groups. Group 1 was the control, group 2 received acrylamide (10 mg/kg, water solution), group 3 received vitamin E (100 mg/kg, intraperitoneal), and group 4 received both acrylamide and vitamin E. After 35 days, spermatozoa from the right cauda epididymis were analyzed in terms of count, motility, morphology, and viability. Sperm DNA integrity and chromatin condensation were assessed by acridine orange (AO), aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining. Results: In acrylamide-treated mice, significantly lower sperm concentration, viability, motility, and testosterone levels were found in comparison with the control and acrylamide+vitamin E groups (p< 0.05). In the vitamin E group, significantly more favorable sperm parameters and testosterone levels were found than in the other groups (p< 0.05). There were also significantly more spermatozoa with less condensed chromatin in the acrylamide-treated mice than in the other groups. Moreover, significantly more spermatozoa with mature nuclei (assessed by AB, CMA3, AO, and TB staining) were present in the vitamin E group than in the control and acrylamide+vitamin E groups. Conclusion: This study revealed the deleterious effects of acrylamide on sperm parameters and sperm chromatin quality. Vitamin E can not only compensate for the toxic effects of acrylamide, but also improve sperm chromatin quality in mice.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.27-33
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• 2021

Objective: The chief outcome of testicular torsion in clinical and experimental contexts is testicular ischemia. Curcumin, a compound with anti-inflammatory and antioxidant properties, has fascinated researchers and clinicians for its promise in the treatment of fertility diseases. Methods: Thirty-five fully grown male mice were randomly classified into five groups: control, sham, testicular torsion, treatment group 1 (testicular torsion+short-term curcumin), and treatment group 2 (testicular torsion+long-term curcumin). Thirty-five days later, spermatozoa from the right cauda epididymis were analyzed with regard to count and motility. Toluidine blue (TB), aniline blue (AB), and chromomycin A3 (CMA3) staining assays were used to evaluate the sperm chromatin integrity. In addition, the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) test was used to assess apoptosis. Results: Treatment group 1 exhibited a remarkably elevated sperm count compared to the testicular torsion group. Additionally, notably lower sperm motility was found in the testicular torsion group compared to the control, treatment 1, and treatment 2 groups. Staining (CMA3, AB, and TB) and the TUNEL test indicated significantly greater testicular torsion in the torsion group compared to the control group (p<0.05). The data also revealed notably lower results of all sperm chromatin assays and lower apoptosis in both treatment groups relative to the testicular torsion group (p<0.05). Significantly elevated (p<0.05) AB and TB results were noted in treatment group 1 compared to treatment group 2. Conclusion: Curcumin can compensate for the harmful effects of testicular ischemia and improve sperm chromatin quality in mice.

• Development and Reproduction
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• v.4 no.1
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• pp.53-59
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• 2000

Endocrine disruptors (EDs) are exogenous chemicals which interfere several aspects of natural hormone properties. EDs with estrogenic activity have been recently reported to cause animal reproductive problems. This study was performed to investigate the effects of bisphenol A (BPA) on the mouse spermatogenesis in vivo. Male ICR mice were orally injected on a daily basis with low dose of BPA 20 mg/kg, high dose of BPA 200 mg/kg, or corn oil (vehicle control) for 7 days, and litter size and weights of body, testis, and cauda epididymis were measured. The level of serum testosterone and the expression of TGF- $\beta$$_1$ mRNA were also analyzed using RIA and RT-PCR, respectively. Also, morphological differences of testes after treatments were examined. Sperm concentration and level of serum testosterone showed a decreasing tendency detected as untreated >corn oil >low >high dose BPA treated mice, although there were no significant statistical differences. Interestingly, in mice treated with a high dose of BPA, partial disappearance of spermatozoa in seminiferous tubular lumen and the expression of TGF-$\beta$$_1$ mRNA were observed. Spermatogenesis was disrupted through TGF-$\beta$ system in the seminiferous tubules, resulting in no development of germ cells. Similarly, the litter size treated with a high dose of BPA was significantly different from that of untreated control group. In conclusion, these results that a high dose of BPA (200 mg/kg) acts as an endocrine disruptor during apermatogenesis in male mice md that there are BPA-specific lesions in the adult male reproductive tract might represent a permanently altered responsiveness to testosterone by BPA in the affected target tissue.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.55-68
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• 2006

Purpose : This study was undertaken to evaluate the effects of the different administration duration of Epimedium Herb extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis and the activities of sperm hyaluronidase. Methods : We used the 2-month-old mice and administered the extract solution of Epimedium Herb 0.3 ml/g/day for 30, 60, 90 and 120 days. The control group was administered the normal saline as the same way. We examined the number of total, motile and normal sperm from the cauda epididymis, the activities of sperm hyaluronidase. Also we observed changes of isolated testis before and after administration of Epimedium Herb extract solutions in the mice. And we compared the testicular tissue especially seminiferous tubules with the control and treated group by histochemical methods. Results : The significant differences were observed in the concentration of total sperm and normality of spermatozoa of the Epimedium Herb extract solution administered groups compared to the control group in 60, 90 and 120 days groups. The significant differences were observed the motility of the Epimedium Herb extract solution administered groups compared to the control group in 60 days group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Epimedium Herb extract solution administered groups compared to the control group, respectively. And the activity of hyaluronidase was significantly increased in the Epimedium Herb extract solution administered groups compared to the control group. Conclusion : This study shows that Epimedium Herb has the beneficial effect on the concentration, morphology and motility of sperm, the testicular tissues and the activities of sperm hyaluronidse in 60 days administration group. We can suggest that Epimedium Herb extract solution be useful for the treatment of male sexual dysfunction and infertility.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.193-198
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• 2008

The current study was designed to extend the technique of spermatogonial transplantation to economically important pig model We evaluated the efficiency of pig to pig transplantation. Donor testis cells were harvested from testes obtained at castration of 10- to 14-day-old boars and were labeled with fluorescent marker(PKH26) before transplantation. The presence of infused dye or labeled pig testicular cells was confirmed in the seminiferous tubules from recipient pig. The most effective procedure of intratubular germ cell transfer was to insert an fine needle ($21{\sim}25$ gauge) through the cauda epididymis and testis into the rete testis under ultrasound guidance. Infusion of $5{\sim}7ml$ of dye solution or cell suspension could fill the rete and up to 50% of seminiferous tubules of 14-week-old boars. Testis were examined for the presence and localization of labeled donor cells immediately after transplantation and labeled donor cells were found in numerous seminiferous tubules from recipient pig testes. These results indicate that germ cell transplantation is feasible in recipient pig testis. This study represents successful spermatogonial transplantation between individual animals in a livestock species.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.62-76
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• 2006

Purpose : These studies were undertaken to evaluate the effects of the Cynomorii Herba (CH) on the spermatogenic abilities such as the concentration, motility and morphological normality of sperm from the testis, and the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Materials and Methods : We used the 2-month-old mice and administered 0.2ml extract solution of CH in the 0.1mg/ml, 1mg/ml, 10mg/ml and 100mg/ml once a day for 60days. The control group was administered the distilled water in the same way. After the administration of extract solution, we examined the number of total, motile and normal sperm from the cauda epididymis, the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. We observed the histological changes of isolated testis and compared to the testicular tissue especially seminiferous tubules between control and CH groups by histochemical method. Results : The concentration of total sperm and the motility of spermatozoa were significantly increased in the 1mg/ml, 10mg/ml and 100mg/ml CH groups, especially in 10mg/ml group, compared to the control group. The significant differences were observed in the normality of spermatozoa of the CH groups compared to the control group. In the histolocal analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the CH groups compared to the control group. Also, the activity of hyaluronidase was significantly increased in the CH groups compared to the control group. In the antioxidant activity analysis, the activities of testicular peroxidase and testicular catalase were significantly increased in the CH groups compared to the control group, respectively. Conclusion : This study shows that CH has the beneficial effect on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. We can suggest that CH extract solution be useful for the treatment of male sexual dysfunctions and infertility.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.1
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• pp.16-31
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• 2007

Purpose: This study was undertaken to evaluate the effects of the different administration duration of Cynomorii Herba extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis and the activities of sperm hyaluronidase. Methods : We used the 8-week-old ICR mice and administered 0.3mg/g extract solution of Cynomorii Herba once a day for 30, 60, 90 and 120 days. The control group was administered the normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis. And we compared the testicular tissue especially seminiferous tubules between control and treated groups by histochemical methods. At the end we observed the difference of sperm hyaluronidase activities between control and treated groups. Results : The significant differences were observed in the concentration of total sperm, the motility and normality of spermatozoa of the Cynomorii Herba extract solution administered groups compared to the control group in 60 and 90 days groups. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Cynomorii Herba extract solution administered groups compared to the control group, respectively. Also, the activity of hyaluronidase was significantly increased in the Cynomorii Herba extract solution administered groups compared to the control group. Conclusion : This study shows that Cynomorii Herba has the beneficial effect on the concentration, morphology and motility of sperm especially in 60 and 90 days administration group. We can suggest that Cynomorii Herba extract solution be useful for the treatment of male sexual dysfunctions and infertility.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.3
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• pp.17-31
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• 2005

Purpose : These studies were undertaken to evaluate the effects of the administration of different concentrated Morindae officinalis Radix extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis and the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Methods : We used the 2-month-old mice and administered the extract solution of Morindae officinalis Radix in the different concentration once a day for 60 days. The control group was administered the normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis, the activities of sperm hyaluronidase, testicular peroxidase and testicular catalase. Also we observed changes of isolated testis before and after administration of Morindae officinalis Radix extract solutions the mice. And we compared to the testicular tissue especially seminiferous tubules between control and treated group by histochemical methods. Results : The significant dose-dependent differences were observed in the concentration of total sperm, the motility and normality of spermatozoa of Morindae officinalis Radix extract solution administered groups compared to the control group, respectively. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Morindae officinalis Radix extract solution administered groups compared to the control group, respectively. Also, the activity of hyaluronidase was significantly increased in the Morindae officinalis Radix extract solution administered groups compared to the control group. In the antioxidant activity analysis, the activity of testicular peroxidase was significantly increased in the Morindae officinalis Radix extract solution administered groups compared to the control group, respectively. Conclusion : This study shows that Morindae officinalis Radix has the beneficial effect on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase and testicular peroxidase. We can suggest that Morindae officinalis Radix extract solution be useful for the treatment of male sexual dysfunctions and infertility.

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.237-244
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• 1990

These studies were carried out to investigate the effects of the follicles size, hormone supplementation, semen types and capacitation methods on in vitro maturation and fertilization of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean Native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, ECS, BFF and MCC for 24~48hrs. in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 18~20hrs. with motile capacitated sperm in the TCF(Tyroide calcium-free) solution containing 200$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The oocytes classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 61.4%, 12.1%, 19.2%, 4.2% and 3.0% of the total oocytes recovered, respectively. The maturation and fertilization rate of the A, B, C class follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS were 89.1%, 78.0%, 52.6% and 78.1%, 66.1%, 33.3%, respectively. 2. The average number of the follicular oocytes recovered from follicles size, 1~2mm, 3~5mm and above 5mm in dimeter were 67, 98 and 63, respectively. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium were 56.7%, 82.5%, 46.0% and 44.8%, 71.4%, 28.6%, respectively. 3. The maturation and fertilization rate of follicular oocytes, cultured in the TCM-199 medium supplemented with 5%, 10%, 15%, 20% FCS and hCG, HCG, $\beta$-estradiol were 76.0%~82.3% and 26.2%~70.0%, and those values were higher the supplementation of the hormone than the non-supplementation. 4. The fertilization and cleavage rate of the follicular oocytes, inseminated with spermatozoas of epididymis cauda, neat and frozen semen were 63.3%, 73.3%, 70.0% and 32.7%, 37.8%, 38.3, respectively. 5. The fertilization and cleavage rate of follicular oocytes, fertilized with capacitated spermatozoas by heparin, BFF and HIS methods were 70.0%, 53.8%, 34.2% and 38.3%, 23.1%, 17.1%, respectively. And the fertilization and cleavage rate were higher method of heparin than other methods.r methods.