• Development and Reproduction
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• v.19 no.4
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• pp.209-215
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• 2015

Red spotted grouper, Epinephelus akaara is a popular aquaculture species in many Asian countries. This species is a protogynous hermaphrodite that first differentiates into female and changes to male later. Due to this reproductive characteristic, stable supply of male and female gametes is a key to the success of seed production in this species. Thus, understanding early gonadal differentiation is required to develop effective sex control techniques. Red spotted grouper were reared in indoor tanks and sampled every 5 days from 40 days post-hatch (DPH) to 130 DPH. Changes of gonadal tissues were examined and analyzed by means of histology. A pair of gonadal primordium has already existed underneath the kidney in the posterior part of the body cavity at 38 DPH when this study began. Gonadal primordia of 38, 40 DPH consisted of germ cells surrounded by a few somatic cells. The blood vessel was observed in the gonadal primordium at 45 DPH. The number of somatic cells and size of gonadal primordium increased age-dependently up to 60 DPH. Formation of ovarian cavity was obvious by two protuberant aggregations of somatic cells at 65 DPH. Completed ovarian cavity and oogonia were first observed in the gonad of one fish sample at 105 DPH. Based on these histological observations, it can be suggested that induction of primary male differentiation could be more successfully applied at around 60 DPH in this species.

• Molecules and Cells
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• v.37 no.1
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• pp.31-35
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• 2014

Induced pluripotent stem cells (iPSCs) are capable of unlimited self-renewal and can give rise to all three germ layers, thereby providing a new platform with which to study mammalian development and epigenetic reprogramming. However, iPSC generation may result in subtle epigenetic variations, such as the aberrant methylation of the Dlk1-Dio3 locus, among the clones, and this heterogeneity constitutes a major drawback to harnessing the full potential of iPSCs. Vitamin C has recently emerged as a safeguard to ensure the normal imprinting of the Dlk1-Dio3 locus during reprogramming. Here, we show that vitamin C exerts its effect in a manner that is independent of the reprogramming kinetics. Moreover, we demonstrate that reprogramming cells under 2i conditions leads to the early upregulation of Prdm14, which in turn results in a highly homogeneous population of authentic pluripotent colonies and prevents the abnormal silencing of the Dlk1-Dio3 locus.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.385-391
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• 2002

Panax ginseng (family- Araliaceae) is a native plant of Korea and has been used for past several years among oriental people. To evaluate the radioprotective potential of P. ginseng on the formation of giant cells in the testis of Swiss albino mice, the animals were divided into four groups: -(I)-Only vehicle was administered. (II)P. ginseng treated group: -The animals received 10 mg/kg body weight P. ginseng root extract (in DDW) i.p. continuously for 30 days. (III) Irradiated group: -The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 ems. (IV) Combined treatment group: -Animals were given P. ginseng extract for four days and on fourth day they were irradiated to 8 Gy gamma radiation after 30 minute of extract administration. The animals of these three groups were autopsied on day 1,3, 7, 14 and 30 days. In ginseng treated group, active spermatogenesis was observed without any toxic effect. Histopathological studies of irradiated group (II) revealed reduction in germ cell count, loss of sperms and formation of multinucleated giant cells on day 7th. These giant cells were formed by round nuclei of early or late spermatids. In combination group (III), although germinal epithelium was still disorganized with loss of cells in few tubules, but no giant cell formation was observed. In order to know the mechanism of radioprotection of ginseng, LPO and GSH were estimated. It was observed that pretreated irradiated animals showed inhibition of LPO and increase in GSH. Thus the present study suggests ginseng protects male gonads. This may be attributed to the inhibition of LPO and increase synthesis of GSH byginseng.

• 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.1
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• pp.19-30
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• 2024

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.236-245
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• 2016

This study investigated the anti-inflammatory effects of wheat germ oil (WGO) on RAW 264.7 cells. It was shown that WGO had no cytotoxicity against the treated cells or negative effect on their proliferation. WGO suppressed nitric oxide (NO) secretion considerably and had inhibitory effects on the production of LPS-induced NO and pro-inflammatory cytokines (IL-6, TNF-α, and IL-1β). In particular, the IL-6 and TNF-α inhibition activities were over 90% at 100 μg/ml concentration of the oil. WGO also inhibited the LPS-induced expression of cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor-kappa B (NF-κB), and reduced the expression of phosphorylated ERK and JNK. Moreover, the croton-oil-induced edema in mouse ears was reduced by WGO, and no mortalities occurred in mice administered 5,000 mg/kg body weight of WGO over a 2-week observation period. In conclusion, these results provide evidence for the anti-inflammatory effect of WGO that likely occurs via modulation of NF-κB and the JNK/ERK MAPK signaling pathway.

• The Korean Journal of Malacology
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• v.31 no.4
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• pp.273-277
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• 2015

This study was conducted to provide the reproductive biological information and basic data on the artificial sex control of Haliotis discus hannai. The morphological sex differentiation process of H. discus hannai could be classified into following five phases: 1) formation of gonad outer membrane (FGOM) (${\leq}SL\;10.0{\pm}1.0mm$), 2) primordial germ cells (PGCs) appearance in the connective tissue between intestine and hepatopancreas (PAC), and formation of gonadal cavity (FGC) (SL $15.0{\pm}2.0mm$), 3) PGCs appearance in the epithelial layer of gonadal cavity (PAG) (SL $18.0{\pm}2.0mm$), 4) formation of gametogenic follicle and appearance of early oocytes and spermatogonia (FGOC) (SL $21.0{\pm}2.0mm$), 5) morphological sex differentiation (MSD) (${\geq}SL\;23.0{\pm}2.0mm$). From histological analysis sex differentiation rate in SL 24.1-25.0 mm of H. discus hannai was 90.0% and sex ratio (female : male) was 1:0.8.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.38-38
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• 2001

Spermatogonial stem cells은 sperrnatogenesis에서 중요한 역할을 하며, 곡세정관의 기저막에 위치하고 있는 것으로 알려져 있다. 그러나, 그 동안 이 세포에 특이하게 발현되는 marker가 거의 알려져 있지 않아 spermatogonial stem cell의 연구에 많은 어려움을 가져왔다. 최근 일반적인 stem cell이 갖는 특성 중, 기저막과 상호작용을 하는 surface protein으로 integrin이 존재한다는 사실을 이용하여, anti-$\alpha$$_{6}$/ 또는 anti-$\beta$$_1$ integrin항체로 germ cell을 선발하여 정소에 이식한 결과, 높은 효율로 이식세포유래의 정자발생이 가능하다는 결과가 보고되었다 (Shinohara et al., 1999). 한편, 항암제의 일종인 busulfan을 마우스에 투여(40mg/kg)한 후 4-5주가 경과하면 세정관의 기저막에 위치하는 spermatogonia를 제외하고 대부분의 생식세포는 소멸한다 본 실험의 목적은 이러한 사실들을 이용하여 spermatogonial stem cell의 특성을 밝히고, 이 생식세포를 보다 간편하고 손쉽게 선발할 수 있는 시스템을 확립하는데 있다. Busulfan처리 후 5주가 경과된 마우스와 정상적인 13주령의 마우스 testis로부터 세포를 분리한 후 FITC-conjugated anti-IgG를 이용한 면역형광법으로 측정.분석한 결과, 형광표식된 세포비율이 대조군과 비교하여 busulfan을 처리한 경우에서 유의적인 증가를 보였다.(17$\pm$3.8%. 0.7$\pm$0.3% busulfan vs control). 또한, IgG와 결합한 단백질이 존재하는 이들 세포들은 곡세정관의 기저막을 따라 위치하며, 단백질과 복합체를 형성한 IgG는 anti-Ig $G_{2a}$와 반응하지 않는다는 사실을 관찰했다. 이러한 IgG 복합체를 형성한 세포들의 특성을 이용하여, IgG와 반응을 하지 않는 것으로 확인된 이차 항체인 an1i-Ig $G_{2}$와 일차 항체인 anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체를 이용하여 측정.분석하였다. Busulfan을 처리한 마우스 정소에서 분리한 세포를 다시 laminin으로 코팅된 dish에서 선발.회수해서, anti-lgG, anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체로 각각 표식된 세포비율을 비교하였다. Laminin으로부터 선발.회수한 세포에서는 IgG복합체가 $\alpha$$_{6}$ 또 는 $\beta$$_1$integrin과 거의 같은 수준에서 높은 비율로 표식되었다. 결론적으로, busulfan에 의해 유도된 IgG와 결합가능한 단백질은 $\alpha$$_{6}$나 $\beta$$_1$ integrin과 마찬가지로 immunoglobulin G를 이용하여 spermatogonial stem cell의 선발을 가능하게 했다. 따라서, busulfan처리시 IgG는 미분화된 정조세포의 선발을 위한 하나의 marker로서 사용가능함을 시사한다.다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.1
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• pp.44-50
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• 1996

Sex differentiation in the rockfish, Sebastes schlegeli, was studied by using a histological method for the appearance of primordial germ cell, formation of primitive gonad and differentiation of female and male. The primordial germ cells were buried under fibrous mesenchymal tissue between gut and mesonephric duct of pre-larva with a total length (TL) of 6.3 mm at 2 days after parturition. In juvenile of TL $5.2\~5.9cm$ at 65 days after parturition, the gonad composed of a large number of genial cell and formed of cavity along the lateral side of the gonad, differentiated to the ovary. At this time, the gonad formed seminiferous tubules by somatic cells, differentiated to the testis. In juvenile of TL $7.0\~7.2cm$ cm at 115 days after parturition, gonads divided into testis contained pigment cell and ovary absent pigment cell. S. schlegeli differentiated directly into male or female without an intermediate female phase at early indifferentiated stage. Therefore, S. schlegeli belongs to the differentiated type of gonochoristic teleosts. At 350 days after parturition, sex ratio was approximately 1 : 1(p>0.05).

• Development and Reproduction
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• v.2 no.1
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• pp.45-52
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• 1998

An enhncer detector line(EDL) having P[1ArB] insertion in X chromosome with expression of reporter gene (lacZ) in the polar cells and border cell of egg chamber was established and used to monitor the differentiation and migration of border cells during the oogenesis of Drosophila. differentiation of border cell from the anterior polar follicle cells was evident in stage-9 egg chamber of EDL149 which was characterized by migration of columnar follicle cells toward posterior of egg chamber surrounding the oocyte. Migration of border cells was observed in the stage-9 and -10 egg chambers. \beta -galactosidase activities were rapidly increased during the first 4 days after eclosion, and it coincided with the timing of border cell differentiation in the ovary during adult life. Homozygote of EDL149 showed some retardation of border cell migration , resulting absence of migration of some border cells in the anterior part of egg chamber or delayed migration of some border cells in the stage-10 egg chamber. These results suggest that the P[1ArB] of EDL149 is inserted at the locus of the structural gene required for the border cell migration. In addition to the expression in egg chambers, lacZ expression was also detected in the meiotic germ cells of testis and antenna, suggesting the possible requirement of the trapped gene function in these organ. this EDL and enhancer trapped gene might be useful for the study of developmentally regulated cell migration.