• Archives of Pharmacal Research
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• v.24 no.4
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• pp.360-366
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• 2001

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various functions such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. Among all ganglio-series gangliosides, GM3 has the simplest carbohydrate structure, and has been shown as a major gangliosides, in male reproductive system. To study GM3 distribution in the seminiferous tubule and epididymis, frozen sections were stained with specific monoclonal antibody (MAb) against ganglioside GM3. In the seminiferous tubule of testis, pachytene spermatocytes and spermmatids expressed ganglioside GM3, but not in spermatogonia and sertoli cells. Spermatogonia and sertoli cells near the basement membrane were negatively reacted to anti-GM3. In the epididymis, GM3 was expressed only in some interstitial cells. Taken togethers, these results suggest that the expression of ganglioside GM3 in rat seminiferous tubule and epididymis is spatio-temporally regu lated during spermatogenesis.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.213-218
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• 2015

In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

• The Korea Journal of Herbology
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• v.24 no.2
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• pp.93-98
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• 2009

Objectives : Previously we reported that the roots of Panax ginseng C.A. Meyer (Araliaceae) increased sperm count and motility. also induced spermatogenesis via cAMP-responsive element modulator(CREM) activation in rat testes. In this study, for the first step of spermatogenesis in germ cell lines, the antioxidant activity of Panax ginseng were examined in mouse GC-1 spermatogonia cells. Methods : The extract was studied on diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, GC-1 cell viability by a modified MIT assay. H202-induced cytotoxicity by MIT assay and lipid peroxidation by malondialdehyde (MDA) formation. respectively. Results: The results showed that the extract scavenged DPPH radical with the IC50 being 0.631 mg/mi. The extract at concentrations of 5, and 10, 50, 100, 250 ${\mu}$g/mi increased GC-1 cell viability significantly(p < 0.05, and p < O.O1). Hydrogen peroxide-induced cytotoxicity (73.8%, p < O.O1) was blocked by the extract at concentrations of 50, and 100, 250, 500 ${\mu}$g/ml significantly (p < 0.05, and p < O.O1). The extract at concentrations of 10. and 50 ${\mu}$g/ml decreased the MDA formation on hydrogen peroxide-induced lipid peroxidation. Conclusions : In conclusion, the extract of Panax ginseng has potent antioxidant activity and increases the survival rate of GC-1 spg cells against $H_20_2$-induced cytotoxicity.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.145-152
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• 2024

Background: Despite its anticancer activity, cisplatin exhibits severe testicular toxicity when used in chemotherapy. Owing to its wide application in cancer therapy, the reduction of damage to normal tissue is of imminent clinical need. In this study, we evaluated the effects of catechin hydrate, a natural flavon-3-ol phytochemical, on cisplatin-induced testicular injury. Methods: Type 2 mouse spermatogonia (GC-1 spg cells) were treated with 0-100 μM catechin and cisplatin. Cell survival was estimated using a cell proliferation assay and Ki-67 immunostaining. Apoptosis was assessed via flow cytometry with the Dead Cell Apoptosis assay. To determine the antioxidant effects of catechin hydrate, Nrf2 expression was measured using qPCR and CellROX staining. The anti-inflammatory effects were evaluated by analyzing the gene and protein expression levels of iNOS and COX2 using qPCR and immunoblotting. Results: The 100 μM catechin hydrate treatment did not affect healthy GC-1 spg cells but, prevented cisplatin-induced GC-1 spg cell death via the regulation of anti-oxidants and inflammation-related molecules. In addition, the number of apoptotic cells, cleaved-caspase 3 level, and BAX gene expression levels were significantly reduced by catechin hydrate treatment in a cisplatin-induced GC-1 spg cell death model. In addition, antioxidant and anti-inflammatory marker genes, including Nrf2, iNOS, and COX2 were significantly downregulated by catechin hydrate treatment in cisplatintreated GC-1 cells. Conclusions: Our study contributes to the opportunity to reintroduce cisplatin into systemic anticancer treatment, with reduced testicular toxicity and restored fertility.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.361-371
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• 2000

It has been recently reported that 2-bromopropane (2-BP) induces male reproductive toxicity in both human and experimental animals. However, delayed effects of 2-BP on male reproductive system have not been investigated in detail. The present study was conducted to investigate the testicular toxicity of 2-BP and to determine the recovery of normal spermatogenesis in Sprague-Dawley rats. Male rats aged 5 weeks were administered 1,000mg/kg 2-BP by gavage daily for 4 weeks and sacrificed sequentially at 1, 2, 3, 4 and 12 weeks after initiation of 2-BP treatment. Testicular toxicity was evaluated qualitatively by histopathological examinations and quantitatively by reproductive organ weights, spermatid head count, and repopulation index. In the 2-BP treated rats, the body weights was significantly suppressed and the weights of testes and epididymides were also decreased in a time-dependent manner. On histopathological examination, spermatogonia in stages I-VI and preleptotene and leptotene spermatocytes in stages VII-IX were strongly depleted at 1 week of dosing. Spermatogonia were depleted extensively in all spermatogenic stages at 2 weeks. Continuing with the evolution of spermatogenic cycle, zygotene spermatocytes, pachytene spermatocytes, and round spermatids were sequentially depleted at 2, 3, and 4 weeks of dosing due to the depletion of their precursor cells. Vacuolization of Sertoli cells and spermatid retention were also observed at all time points, suggesting that 2-BP induced Sertoli cell dysfunction. At 12 weeks, after 8 weeks recovery, most of the tubules appeared severely atrophic and were lined by Sertoli cells only. Leydig cell hyperplasia in the interstitial tissue was also found. In addition, dramatic reductions in the number of spermatid heads and repopulation index were observed, indicating that 2-BP-induced testicular injury is irreversible. These results indicate that 4 weeks repeated-dose of 1,000mg/kg 2-BP results in a progressive germ cell loss due to the depletion of spermatogonia followed by long-term testicular atrophy in SD rats.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.76-76
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• 2003

Eleven wrasse species inhabit the coastal waters of Jeju Island, Korea. They are the target of leisure fishing and are considered good eating. We investigated the distribution of standard length (SL) by sex of wrasse in Jeju coastal waters for Halichores poecilopterus, H. tenuispinis, Pseudolabrus japonicus, and Pteragogus flagellifera. A cross-section of the ovary showed the ovarian cavity and ovarian lamellae containing oocytes. A cross-section of the testis showed many lobules containing spermatogonia and spermatocytes. A cross-section of a gonad undergoing sex reversal showed the regression or reduction of oocytes and some spermatocytes located in the ovarian lamellae. A cross-section of a sex-reversed testis showed the primary structure of the ovary, with spermatocytes distributed in the epithelium of the lamellae, and reformed seminiferous ducts in the basement lamellae. (중략)

• Biomedical Science Letters
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• v.10 no.3
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• pp.275-283
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• 2004

The annual changes in testis weight and diameter of seminiferous tubules, and the seminiferous epithelium cycle of Tamias sibiricus were studied by light microscope. Testis weight and diameter of seminiferous tubule are significantly increased from January to July, and decreased rapidly to the size from August to December. Spermatogenesis occurs from January to July, and spermatocytogenesis are produced from August to December. The cycle of the seminiferous epithelium was divided into 12 stages during the development of spermatids as a changes of the nucleus and acrosomal structure, presence and/or absence of residual body, appearance and/or absence of sperm tail and meiotic figure and spermiation. The dark type spermatogonia (Ad) are appeared in all stages (I ~ XII), and the spermatids of step 10 are observed at I, II, X and XII stages. The spermatids of step 11 are appeared in III and IV stages, only the step 12 spermatid observed in V stage.

• Biomedical Science Letters
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• v.17 no.3
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• pp.239-243
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• 2011

Radiation exposure may cause tissue damage by ionized or excited atoms. The molecules are transformed from water or they directly interact with cells. Antioxidants such as vitamin E, vitamin C are used to reduce the interference of radiation. Especially, vitamin E inhibits the formation and accumulation of lipid and oxidation and the activity of drug-metabolizing enzymes. After irradiation of 3 Gy to the testis, the entire shape of testis was partly depressed and an irregular vacuole was observed to the edge. The 3 Gy group after treatment with vitamin E showed the low destruction of spermatogonia and partial tissue damage as compared to the 3 Gy group. Vitamin E inhibited the production of vacuoles from the cell death and generated the block to prevent the condensation of the seminiferous tubules.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.7
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• pp.1015-1022
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• 2007

The spermatogonial stem cell (SSCs) is unique in that it is the only cell in the adult male that can contribute genes to a subsequent generation. Permanent modification of the germ cell line may be realized if stem cells could be cultured, transfected with unique genes, and then transplanted into recipient testes. We developed a culture system that supported long-term viability of SSCs. We used a retrovirus vector (pMSCV including ${\beta}$-galactosidase) to stably transfect spermatogonia following long-term culture using the system developed. Expression of the reporter gene ${\beta}$-galactosidase controlled by the retroviral vector was stable in long-term cultured SSCs. We confirmed the retroviral-mediated ${\beta}$-galactsidase gene could be expressed in germ cells in recipient mice following SSCs transplantation.

• Journal of Veterinary Clinics
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• v.16 no.2
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• pp.448-453
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• 1999

The three-dimensional structure of the Sertoli cell in the Korean native bull was investigated by scanning electron microscopy. Morphologically, four types of Sertoli cell processes were evident: 1) sheet-like processes, 2) sleeve-like processes, 3) bough-like processes and 4) finger-like processes. The sheet-like processes rested upon more than half of the surface of each spermatogonia, spermatocyte and spermatid. Sleeve-like processes, bough-like processes and finger-like processes are observed in the middle and apical portion of seminiferous tubule. All Sertoli cell processes are originated from Sertoli cell column. Just before spermiation, the apical sheet-like processes are shifted from their position at the spermatid head, and bough-like processes covered the disengaged residual body, after which the residual body was no longer evident in the tubule. Though the mechanism for this elimination is not known, the process suggests a reciprocity between the Sertoli and germ cells.