• 한국가축번식학회지
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• 제11권1호
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• pp.33-41
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• 1987

The cycle of the seminiferous epthelia in the testis of matured Korean Native Cattle was divided into eight stages. The results were summarized as follows: 1. Type A spermatogonia a, pp.ared twice as many at stage 2 as compared to stage 1, while maximum numbers were the average of 2.8 at stage 2. The intermediate and Type B spermatogonia were found during the stage 3 to 8, stage 6 to 8, respectively. The leptolene primary spermatocytes were not observed during the stage 5 to 7, while the pachytene primary spermatocytes were shown the least in number at stage 4, the secondary supermatocytes could be seen only at stage 4 and the round spermatids were not observed at stage 3, 4. 2. The relative frequencies of the eight stages of the cycle of the seminiferous eptithelia were 24.9, 14.2, 19.0, 6.3, 3.7, 7.9, 10.3 and 13.9%, respectively. 3. Some of the nuclei of Sertoli cells transformed from the "parallel" type to the "perpendicular" type. This evolution took place from stage 1 to 5, when the number of "perpendicular" type nuclei reached a peak and the number was decreased in the rest of the stages.sed in the rest of the stages.

• 대한수의학회지
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• 제33권1호
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• pp.23-36
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• 1993

To investigate the cycle and relative frequences and the fine structure of seminiferous epithelia in mature Jindo dogs, histologic study was performed. The results obtained were summarized as follows; 1. Type A spermatogonia appeared approximately 1.6 times as many at stage II as compared to stage I while type In spermatogonia appeared small amount in stage III, IV and V. type B spermatogonia were found during the stage VI to VIII, though not detectable during stage I to V. The type B spermatogonia divided at stage VII to produce the preleptotene primary spermatocytes at stage VIII. The number of primary spermatocytes of the leptotene phase markedly increased during stage I to II, and the primary spermatocytes of the pachytene phase were shown the least in number at stage IV. The secondary spermatocytes could be seen only at stage IV. 2. The relative frequencies of each stage from stages I to VIII of the cycle of seminiferous epithelia were 31.6, 11.9, 10.0, 3.2, 8.2, 10.1, 11.7 and 13.2% respectively. 3. On electron microscopic observations, acrosomal vesicle of spermatids appeared larger though the bulk of germ cells were the morphologically same as those of the other animal species. Thread line structures light microscopically observed in the cytoplasm of Sertoli cell were the longitudinal orientation of mitochondria.

• 대한방사선기술학회지:방사선기술과학
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• 제13권2호
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• pp.51-51
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• 1990

This study was undertaken to observe the effects of $^{60}Co\;{\gamma}-irradiation$ on the cell of spermatogenic epithelium in the testis of the chicken. 16-week-old chicken were provided as an experimental group and compared with control group. The experimental group was divided into a single irradiation (800, 1000, 1200 rads) and into three partial irradiation group (800/3, 1000/3, 1200/3 rads). The morphological changes of epithelial cell of the testis were observed by means of hematoxyline and eosin stain. Microstructure of spermatocyte and sperm was observed by means of semithin section of electron microscopic specimen. The results obstained are summerized as follows. 1. Spermatogonia and sertoli cells were found to be isolated from the basal membrane of seminiferous tubules as dose of $^{60}Co\;{\gamma}-irradiation$ was increased. 2. Spermatocytes of pachytene stage were seperated from the cytotplasmic process of sertoil cell in case of 1000 rads of $^{60}Co\;{\gamma}-irradiation$. 3. Normal arrangement of the cell of spermatogenic epithelium was found in control group and only the partial irradiation group of 800 rads. Vaculation in the seminiferous was pronounced in case of a single irradiation group of 800 rads, but the irradiation group of 1000 rads and 1200 rads were found to be damaged severely in both a single and a partial dose.

• Asian-Australasian Journal of Animal Sciences
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• 제22권2호
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• pp.187-193
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• 2009

The present study identified the favorable conditions for isolation, enrichment and in vitro culture of highly purified, undifferentiated pig spermatogonial stem cell (SSC) lines that proliferate for long periods of time in culture. The colonies displayed morphology similar to miceSSC and were positive for markers of SSC (PGP9.5), proliferating germ cell (PigVASA), pre-meiotic germ cell (DAZL) and pluripotency (OCT4, SSEA-1, NANOG, and SOX2) based on immuno-cytochemistry and RT-PCR. The purity of these colonies was confirmed by negative expression of markers for sertoli cell (GATA4 and SOX9), peritubular myoid cell (${\alpha}$-SMA), differentiating spermatogonial and germ cells (c-KIT). The colonies could be maintained with undifferentiated morphology for more than two months and passaged more than 8 times with doubling time between 6-7 days. Taken together, we conclude that pigSSC could be successfully isolated and cultured in vitro and they possess characteristics similar to miceSSC.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.225-230
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• 1999

In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

• 한국수정란이식학회지
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• 제28권2호
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• pp.149-155
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• 2013

Mullerian inhibiting substance (MIS) is a member of the TGF-${\beta}$ (transforming growth factor-${\beta}$) family whose members play key roles in development, suppression of tumour growth, and feedback control of the pituitary-gonadal hormone axis. MIS is expressed in a highly tissue-specific manner in which it is restricted to male Sertoli cells and female granulose cells. The serum levels of MIS in prenatal and postnatal ICR mice were measured using the enzyme-linked immuno-solvent assay (ELISA) using the MIS/AMH antibody. Mice were grouped by age: the significant periods were at the onset of development. During sex organ differentiation, no remarkable difference between female and male foetus MIS serum levels (both<0.1 ng/ml) was observed. However, MIS serum levels in pregnant mice markedly changed (4.5~12.2 ng/ml). After birth, postnatal female and male mice serum MIS levels changed considerably (male: <0.1~138.5 ng/ml, female: 5.3~103.4 ng/ml), and the changing phase were diametrically opposed (male: decreasing, female: fluctuating). These findings suggest that MIS may have strong associations with not only develop-ment but also puberty. For further studies, establishing the standard MIS serum levels is of importance. Our study provides the basic information for the study of MIS interactions with reproductive organ disability, cancer, and the effect of other hormone or menopause. We hypothesise that if MIS is regularly injected into middle-age women, meno-pause will be delayed. We detected that serum MIS concentration curves change with age. The changing phase is different between males and females, and this difference is significant after birth. Moreover, MIS mRNA is expressed during the developmental period (prenatal) and also in the postnatal period. This finding indicates that MIS may play a significant role in the developmental stage and in growth after birth.

• Clinical and Experimental Reproductive Medicine
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• 제30권2호
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• pp.127-133
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• 2003

Objective: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. Methods: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. Results: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. Conclusions: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

• Journal of Ginseng Research
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• 제34권4호
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• pp.327-335
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• 2010

We reported previously that the administration of Korean red ginseng water extract (KRG-WE) protected the guinea pig testis against damage induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (a potent endocrine disruptor). We also found that crude saponin from ginseng was the active ingredient responsible for this protection. Here, we examined the biological role of KRG-WE in an animal model of age-induced dysfunction of spermatogenesis. Twenty-four male Sprague-Dawley (six 2-month-old and eighteen 12-month-old) rats were used. The young and old control groups received only vehicle. The ginseng saponin (GS)- and KRG-WE-treated groups received GS (40 mg/kg body weight/day) and KRG-WE (200 mg/kg body weight/day), respectively, for 4 months. The number of cells, Sertoli cell index, Johnsen's score, and sex hormone levels decreased significantly with age. However, the administration of KRG-WE and GS markedly improved the number of germ cells, seminiferous tubular size, and Johnsen's score in the old rats. Ginseng produced a distinct testicular histological improvement in old rats. KRG-WE and GS elevated testosterone levels, while attenuating the aberrant increase in follicle stimulating hormone and luteinizing hormone levels. Sperm kinematics evaluated by a computer-assisted sperm analyzer demonstrated improvement in the percentage of motile sperm, progressive sperm motility, and curvilinear velocity associated with sperm quality, supporting the beneficial role of red ginseng in senile spermatogenesis. Overall, the total water extract had a more potent effect than the corresponding saponin fraction. In conclusion, Korean red ginseng rejuvenated age-induced testicular dysfunction. Additionally, the total water extract was more potent than the corresponding saponin fraction.

• 생명과학회지
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• 제14권1호
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• pp.72-81
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• 2004

TBTCl(0.1, 0.32, 1, 3.2, 10, 25, 32, 50, 75 및 100 $\mug/l$)을 구피에게 144시간 동안 노출시켜 TBTCl의 구피 생체 내 축적정도 및 구피의 번식과 행동에 미치는 영향에 대해서 조사하였다. 0.1 $\mug/l$ 이상의 모든 TBTCl 실험군에서 $\SigmaBTs\; (TBT:\SigmaBTs)$가 67% 이상으로 매우 높았고, 고농도 실험군일수록 이 비율이 높아져, 고농도의 TBTCl에 노출될수록 구피의 대사능력이 저해됨을 알 수 있었다. 또한 TBTCl은 구피의 번식력을 약화시키고 행동이상을 유발함이 밝혀졌다. 즉, 0.1$\mug/l$의 실험군부터 수컷의 구애행동과 수출관 내 정포의 수가 감소되었고, 0.32∼10 $\mug/l$의 실험군에서는 정소 내 전체 생식세포낭에 대한 정포낭의 수도 점차 감소하여 TBTCl이 정자변태 과정에 영향을 미치는 내분비계 장애물질로서 작용하고 있음이 시사되었다. 더욱이 25 $\mug/l$이상의 고농도군에서는 단기간 내에 치사율이 증가하고, 생식세포가 괴사되고, 정소의 지지세포와 수출관 내의 상피세포가 상해되고, 유영행동 이상이 급증하고, 섭식이 중단되는 등 급성독성이 관찰되어, TBTCl이 구피의 번식 뿐 아니라 생존을 저해하고 있음이 시사되었다.

• Biomolecules & Therapeutics
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• 제4권2호
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• pp.127-137
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• 1996

This study was performed to investigate the toxicity of DA-125 in beagle dogs, an anthracycline antitumor antibiotic. The dogs were administered DA-125 i.v. at 0.0023, 0.0375, 0.15 and 0.6 mg/kg/day, 6 days/week for 26 weeks. At 0.6 mg/kg, all male and female dogs were either sacrificed moribundly or dead during the 26-week treatment. The dogs revealed inactivity, salivation, dark bloody discharge, swelling of the subcutaneous injection site, abscess, and ulceration in the abdominal wall and legs. At 0.15 mg/kg, anorexia, salivation, and swelling of the injection site were observed. The food consumption was decreased with a statistical significance at 6 and 12 weeks treatment in males of 7.6 mg/kg. At 0.0375, 0.15 and 0.6 mg/kg, body weights were decreased significantly in a dose-related fashion after 17 weeks treatment. Total white blood cell counts for male dogs at 0.6 mg/kg were lower than those of control dogs after 13 weeks treatment, which appeared mainly due to decreased neutrophils. At 0.15 mg/kg, testicular atrophy was found in all males by gross pathology and the testicular weights were significantly decreased when compared to those of control males. Microscopically, the testis showed moderate atrophy of the seminiferous tubules and marked decrease in number of spermatozoa in the epididymal tubules. At 0.6 mg/kg, petechia or echymotic hemorrhage was observed in gastrointestinal tract, heart, lungs, and other organs at the necropsy, Marked atrophy of thymus were observed in both males and females. In addition, severe testicular atrophy was noted in all males. Microscopically, gastrointestinal tract showed hemorrhage, epithelial denudation, hypermucus secretion, and atrophy of intestinal villi. Seminiferous tubules of the atrophic testis were lined with Sertoli cells only and devoid of germ cells. Severe oligospermia or aspermia was present in the epididymal tubules. Bone marrow showed marked depletion of hemopoietic cells. In addition, marked atrophy was found in the lymphoid tissue of gastrointestinal tract, various Iymph nodes, and thymus. Injection sites showed marked inflammatory response with necrosis, necrotizing vasculitis, thrombus formation, and ulceration in the skin. According to the present results, no observed effect level appeared to be 0.0375 mg/kg. At 0.15 mg/kg, testis was a target organ, while at 0.6 mg/kg hemopoietic tissue, gastrointestinal tract, and testis were considered to be target organs. At 0.6 mg/kg the test compound seems to inflict a damage on the blood vessels causing hemorrhage in the various organs and tissues.