• Clinical and Experimental Reproductive Medicine
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• v.31 no.2
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• pp.83-94
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• 2004

Objective: To investigate whether GnRH-agonist (GnRH-Ag) using in IVF-ET affects apoptosis of human granulosa-luteal cells and expression of peripheral benzodiazepine receptor (PBR) protein involved in the apoptosis of the cells. Methods: Granulosa-luteal cells obtained during oocyte retrieval were cultured and treated with $10^{-5}M$ GnRH-Ag. Apoptosis of the cells by the treatment was confirmed using DNA fragmentation analysis 24 h after culture. The presence of PBR protein within the cells was examined by immunofluorescence staining and the expression of the protein was analyzed by Western blotting. In addition, it was measured for progesterone and nitric oxide (NO) produced by granulosa-luteal cells after GnRH-Ag treatment. To evaluate the relationship between NO production and PBR expression, sodium nitroprusside (SNP) as a NO donor was added in media and investigated the expression of PBR protein by Western blotting. Results: Apoptosis increased in the granulosa-luteal cells 24 h after GnRH-Ag treatment, whereas the expression of PBR protein significantly decreased. Furthermore, the production of progesterone and nitric oxide (NO) by the cells significantly fell from 12 h after the treatment. In the results of Western blotting after SNP treatment, the expression of PBR protein increased in the treatment with SNP alone to the granulosa-luteal cells, but was suppressed in the treatment with GnRH-Ag and SNP. Additionally, the staining result of PBR protein in the cells showed the even distribution of it through the cell. Conclusion: These results demonstrate that GnRH-Ag treatment induces apoptosis, decreasing expression of PBR protein and NO production in human granulosa-luteal cells. The present study suggests that one of the apoptosis mechanism of human granulosa-luteal cells by GnRH-Ag might be a signal transduction pathway via NO and PBR.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.433-436
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• 2004

Polycystic ovarian syndrome (PCOS) is a very common endocrine disorder in women of reproductive age. There are some evidences that nerve growth factor (NGF) is involved in the pathogenesis of PCOS. In this study, we investigated the effect of Korean red ginseng total saponin (GTS) on the ovarian morphology and NGF expressions in the ovaries, pituitary and hippocampus. The oil control animals were injected with 0.2 ml oil/rat. Animals in estradiol valerate (EV) control group were injected i.m. with 4 mg EV in 0.2 ml oil/rat. The GTS was administered (100 mg/kg) i.p. every other day for 60 days, beginning 1 day after the EV injection. PCO was induced by a single injection of EV (4 mg, i.m.). At day 60, the expressions of NGF were examined by immunohistochemistry. The main findings were as follows; PCO was fully developed with a single i.m. injection of EV, and PCO showed the increased expression of NGF, and GTS administration decreased NGF expressions in the ovaries without affecting pituitary and hippocampus significantly. The present results demonstrate that GTS attenuates PCOS by the stimulation of NGF expression.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.265-274
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• 2009

Objective: MicroRNAs (miR) are known to repress target genes at post-transcriptional level and play important roles in development and maturation of cell. However, the expression profiles of miR during ovarian follicle maturation have not been fully elucidated. Here, we designed this study to investigate the expression profiles of miR in oocytes and granulose cells (G-cells) after in vitro culture according to gonadotropins and adding hCG. Methods: Ovaries from 12-day-old mice (C57BL6) were removed and preantral follicles were isolated and cultured in $20\;{\mu}L$-drop of culture media with supplementation of either rFSH, rLH, or rFSH+rLH. After their full maturation, follicles were incubated with rhCG and rEGF. RNA was isolated from oocytes and G-cells, and real-time PCR were performed with primers of miR known to be expressed in the mouse ovary (mmu-miR-16, -miR-27a, -miR-126, -miR-721). Results: FSH+LH group showed the highest ovulation and MII rates among gonadotropin groups. The profiles of miRs in oocytes and G-cells differed according to gonadotropin groups and adding hCG. The profiles of miRs showed divergent changes between oocytes and G-cells. Conclusion: miR expression profiles are altered by gonadotropins and supplementation of hCG during in vitro maturation of murine follicles. Target gene study must be necessary to validate these findings.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.395-406
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• 2000

The present study was conducted to examine the developmental capacity of mouse oocytes within prenatal follicles cultured various concentrations of FSH and LH and the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc) and cytochrome P450 17 $\alpha$ -hydroxylase (P450)$_{17{\alpha}}$ mRNA, as luteinization and atretic marker, in these culture conditions. In addition, we investigated the concentrations of progesterone and testosterone in culture medium. The developmental potential up to blastocyst of the oocytes grown in vitro was higher in the FSH alone (30.2%) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (28.0%) groups than in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (22.0%). And the mean numbers of cell per blastocyst was higher in the FSH alone (50.9$\pm$26.1) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (51.0$\pm$21.1) groups when compared to the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (45.2$\pm$15.1). The expressions of P450scc and P450$_{17{\alpha}}$ mRNA in the oocyte -cumulus complexes were increased with increasing of LH concentration, and also the secretions of progesterone and testosterone were increased. Especially, in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group, the expression of P450scc and P450$_{17{\alpha}}$ were significantly increased, and the secretion of progesterone and testosterone were significantly increased. Therefore, these data show that gonadotrophins are essential for the in vitro culture of preantral follicles, but that increasing of LH concentration is reduced the developmental capacity of oocytes. The cause of these findings may be due to increasing of progesterone and testosterone secretion by the enhance of P450scc and P450$_{17{\alpha}}$ mRNA expressions, as markers of luteinization and atresia. Conclusively, this study suggest that supplementation of 100 $m\ell$U/$m\ell$ FSH or 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH may be optimal condition for the culture of mouse pre antral follicles.

• Korean Journal of Agricultural Science
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• v.9 no.1
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• pp.303-313
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• 1982

The study was conducted to find out the concentrations of sex hormones and the contents of metabolites in serum, and the changes of weights and tissues on ovary, thyroid gland and adrenal gland according to gestation period in rabbit. The results were summarized as follows: 1. The ovary weights were increased significantly with the time e lapse after gestation and recovered normally at 5 days after parturition. In the histological changes of ovary, the lutein cells were hypertrophied and the secretory granules were increased actively until 3 weeks after gestation, and then a trophied thereafter. 2. The thyroid gland weights at all observation times were higher than those in control group, and the significance was recognized at 1, 3 and 4 weeks after gestation. The histological features of the secretory epithelium were the hyper trophic and columnar condition stimulating the functional state from 1 week after gestation. 3. The adrenal gland weights in experimental group were recognized significantly at 4 weeks after gestation, but showed higher than those in control group at all observation times. The zona fasciculata and zona reticularis of the gland showed the slight hypertrophic condition, but the zona glomeerulosa and adrenal medulla did not find out any particular changes. 4. The serum concentrations of progesterone and LH reached a peak level at 2 weeks and 1 week after geestation respectively, and rapidly began to decline thereafter. 5. The serum concentrations of estradiol-$17{\beta}$ and FSH were not detected below 20.0 pg/ml and 1.3 mIU/ml respectively. 6. The contents of total protein and non-protein nitrogen nitrogen were decreased gradually with the time elapse after gestation, but the significant differences were recognized from 3 weeks. 7. The total lipids were not changed markedly until 3 weeks, but increased significantly at 4 weeks after gestation and at 5 days after parturition. 8. The serum cholesterol tended to be decreased until 3 weeks, but increased at 4 weeks after gestation and at 5 days after parturition. 9. The serum calcium showed a continuous decrease during the gestation period, but the significant differences were recognized at 3 and 4 weeks. The serum phosphorus also had a significant decrease at 4 weeks after gestation.

• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.149-154
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• 2013

The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89~93 in ovulation phase were transferrin, and spot No.94~98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC-E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.

• Korean Journal of Environmental Biology
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• v.35 no.2
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• pp.160-168
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• 2017

Tscherskia triton is widely distributed in Northern China, Korea, and the adjacent areas of Russia. Except its distribution, reproduction, and growth development related to life history, reproductive cycle and reproductive organs of T. triton are rarely studied in Korea. The purpose of this study was characterized the estrous cycle of T. triton captured in Jeju Island in order to provide information to a better information of captive breeding of the species when long-day (16L : 8D) and short-day (8L : 16D) photoperiod. Then, histological study of the ovaries and uterus with five females in each photoperiod was performed. The duration of the estrus cycle was 4~5 days and it showed regular cycle pattern. Results of the vaginal cytology examination showed four characteristic phase of the estrous cycle in long-day photoperiod (16L : 8D): proestrus, estrus, metestrus and diestrus. However, in short-day photoperiod, the diestrus phage of the estrus cycle was maintained from the $6^{th}$ to $12^{th}$ day. In the long-day photoperiod, females had many Graafian follicles and corpus luteums in large ovaries, and developed uterine glands in the thick endometrium. But they had some primary, secondary and tertiary follicles, and undeveloped uterine glands in the thin endometrium during short-day photoperiod. These results were identified difference of the estrus cycle and histological characteristics of reproductive tracts according to the photoperiod. These results are very important clues to the reproductive biology of T. triton, and it will be widely used as date for maintaining biodiversity.

• Korean Journal of Environmental Biology
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• v.20 no.3
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• pp.224-231
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• 2002

Photoperiod (length of light per day) is a major factor in regulating reproductive function in golden hamsters. The information of photoperiod is transmitted to the reproductive endocrine system by melatonin. Thus the effects of melatonin aye investigated in male golden hamsters exposed to photoperiods. Paired testicular weights were markedly reduced in the animals housed in short photoperiod $(SP,\le{12\;hours\;day^{-1})$ and injected with melatonin in the evening, but not in long photoperiod $(LP,\le{12.5}\;hours\;day^{-1})$ and injected with melatonin in the morning. The histological examination of regressed testes showed reduction of tubular lumen diameter including the numbers of cells and Leydig cell number. The mean values of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) were also lowered in the sexually inactive animals than in the sexually active animals. Melatonin receptor was identified by reverse-transcription polymerase chain reaction (RT-PCR) and its expression was examined in various tissues to scrutinize the action site of melatonin. It turned out 309 nucleotides and was definitely expressed in hypothalamus and pituitary including spleen, retina, and epididymis. And gonadotropin releasing hormone (GnRH) gene, which is a key element in regulating reproduction, was identified by RT-PCR but the expression of GnRH was not modified by the treatment of melatonin. Taken together, photoperiod via melatonin indirectly affects reproductive endocrine system, possibly through the release of GnRH, not the synthesis of GnRH.

• Clinical and Experimental Pediatrics
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• v.52 no.1
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• pp.111-118
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• 2009

Puopose : Exposure to dietary phytoestrogens such as genistein during early childhood is a growing public health concern. We examined the effect of early exposure to genistein on sexual maturation in immature rats. Methods : Weaning (3wk-old) Sprague-Dawley female rats were assigned to three groups (n=6 for each): fed by high dose of genistein (100 mg/kg/d), low dose of genistein (10 mg/kg/d) and control group. First vaginal opening (VO) day was observed. Structural alterations in the ovary and uterus were assessed by histologically. Expression of genes of $ER{\alpha}$, $ER{\beta}$, and progesterone receptor (PR) in the ovary and uterus were investigated by RT-PCR. Results : High genistein group had earlier VO than control and low genistein group. Graafian follicles and corpora lutea were observed from the ovary of genistein-treated groups, while primary, secondary follicles and small atretic follicles were observed in the control group. Hypertrophy of luminal and glandular uterine epithelia were found in the genistein-treated groups while poor development of gland and fewer myometrial cell layers were evident in control group. In ovary, the transcriptional activities of $ER{\alpha}$ and $ER{\beta}$ were higher in high genistein group than in controls. In uterus, the transcriptional activities of $ER{\alpha}$, $ER{\beta}$ and PR were higher in low genistein group than in controls. Conclusion : Acute exposure to genistein during the prepubertal period could activate the reproductive endocrine system resulting in the early onset of puberty in female rats. Further clinical investigation on the effect of genistein on the sexual maturation in children is warranted.

• Journal of Embryo Transfer
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• v.18 no.2
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• pp.157-161
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• 2003

The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes in vitro maturation of cats oocytes and development of IVM/IVF embryos. The results were summarized as follows : 1. The fertilization and developmental rate of fresh and salts-stored oocytes with and whithout cumulus cells were 65.7%, 17.1% and 28.6%, 8.6% and 57.1%, 13.3%, 23.3%, 3.3%, respectively. The rate of oocytes with cumulus cells(13.3%∼65.7%) was higher than that of denuded oocytes(3.3%∼28.6%). 2. The fertilization and developmental rate of oocytes recovered from ovaries collected at different stages of the reproductive cycle were 68.9%, 44.4%, 48.9% and 17.8%, 8.9%, 12.8%, respectively. 3. The fertilization and developmental rate of oocytes in vitro cultured at different time of incubation(24, 36 and 48 h) were 66.7%, 46.7%, 48.9% and 17.8%, 11.1%, 8.5%, respectively. respectively. The rate of oocytes incubated 24 h(66.7%) was higher than that oocytes incubated 36 and 48 h(46.7%∼48.9%). 4. The fertilization and developmental rate of oocytes treated activation and non-activation oocytes were 57.4%, 31.4% and 22.9%, 11.4%, respectively. The rate of oocytes treated activation was higher than that oocyte treat non-activation.