• Korean Journal of Fisheries and Aquatic Sciences
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• v.22 no.5
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• pp.266-280
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• 1989

Gonadal maturation of the Korean pomfrets, Pampus echinogaster (Basilewsky) and Pampus argenteus (Euphrasen) were histologically investigated based on the samples captured in the East China Sea from January 1987 to December 1988. Gonadosomatic index (GSI) of P. echinogaster began to increase from March, and reached maximum between May and July. It began to decrease from July and reached mini-mum between August and February. P. argenteus had a similar cycle, however, P. argenteus has higher values in April than P. echinogaster. Hepatosomatic index (HSI) were positively related to GSI. HIS of P. echinogaster and P. argenteus reached maximum in $April\~July$ and $April\~August$, respectively, Fatness coefficient of two Pampus species were low in the summer, and high in the winter. Ovary is of saccular structure, and testis is of lobular structure. From February, the early oocyte (ca. $100\mu$ in diameter grows) rapidly at the germinal epithelium of ovarian sacs. From March to April the oocytes grew up to cu $400\~500\mu$ in diameter. At this stage, the yolk globules are accumulated rapidly in the cytoplasmic layer. From May, the oocytes roached ca. $650\~850\mu$ in diameter, and they are spawned in $May\~July$. After spawning the residual follicles and remained ripe eggs degenerate. From February, spermatogonia grows into spermatocyte on the epithelium of the testicular lobuli. From May, spermatozoa appeared and spawning occurs. After spawning, the epithelium is thickened and the remained spermatozoa degenerate. Annual reproductive cycle of two Pampus species could be divided into four successive stages: Growing stage ($March\~April$), Mature stage ($April\~May$), Ripe and spent stage ($June\~July$) and Recovery and resting stage ($August\~January$). Absolute fecundity of P. echinogaster was $9,441\~135,294$, and that of P. argenteus was $50,678\~221,894$. Absolute fecundity of two Pampus species were positively related to body length and total weight. Relative fecundity was positively related to body length, while it was reversely related to total weight. The increasing rate of absolute fecundity of P. echinogaster was lower than P. argenteus. In P. echinogaster half of female and male reached first maturity at body length of $15.0\~$17.9cm and $12.0\~14.9cm$, respectively. All of females and males reached first maturity at body length of $18.0\~20.9cm$ and $21.0\~23.9cm, respectively. In P. argenteus all of females and males reached first maturity at body length of 18.6cm and 16.7cm$, respectively.

• Journal of Aquaculture
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• v.4 no.2
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• pp.97-110
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• 1991

This study was undertaken to understand the seasonal changes of hormone levels such as luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol-$17{\beta}$, serum components such as albumin, total protein and triglycerides, and GSI of rainbow trout (Onco-rhynchus mykiss) under the normal-light circumstances. LH levels were the highest in November whereas FSH levels were decreased progressively from October to February. Serum GTH, such as LH and FSH, levels were low immediately after egg-stripping and were significantly increased in the summer in association with the initiation of vitellogenesis. Serum albumin, total protein, and triglycerides were significantly increased in August in association with initiation of vitellogenesis and were peak in November prior to egg-stripping. Oocyte growth, measured by oocyte diameter and GSI, was significantly increased in August and showed the highest value in January. Correlation coefficients of serum LH with FSH and estradiol-$17{\beta}$ were +0.844 and +0.947, respectively. Correlation coefficients of estradiol-$17{\beta}$ with albumin and total protein were +0.634 and +0.859, respectively. Correlation coefficients of serum estradiol-$17{\beta}$ with triglycerides and GSI were +0.673 and +0.694, respectively. Probably by positive feedback mechanism, LH, FSH and estradiol-$17{\beta}$ appeared to stimulate to liver to secrete albumin, total protein and triglycerides for vitellosenesis.

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.119-124
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• 2006

The present study compared the developmental potential of cloned porcine embryos with mesenchymal stem cells (MSCs), fetal fibroblasts (FFs) and cumulus cells (CCs) by assessing the cleavage and blastocyst rate, total cell number, inner cell mass (ICM) ratio and apoptosis. MSCs were isolated by ficoll gradients from femur of -6 month old female pig, and maintained for primary cultures. FFs from a female fetus at ${\sim}30$ day of gestation were established, and CCs were obtained from cumulus oocyte complexes (COCs) aspirated from $3{\sim}6$ mm follicles in diameter. Donor cells at $3{\sim}4$ passage were employed for nuclear transfer (NT). COCs were matured and fertilized in vitro(IVF) as control. Cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs, FFs and CCs ($82.7{\pm}8.9%\;vs\;70.6{\pm}5.4,\;68.7{\pm}5.1\;and\;63.4{\pm}5.6%$, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs ($24.5{\pm}2.8\;and\;20.4{\pm}8.3%$) did not differ, but were significantly (P<0.05) higher than NT derived from FFs and CCs ($10.6{\pm}2.7\;and\;9.8{\pm}2.1%$). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs ($35.4{\pm}5.2\;and\;0.40{\pm}0.09%$, respectively) were significantly (P<0.05) higher than those from FFs and CCs ($24.9{\pm}6.2%\;vs\;0.19{\pm}0.16,\;23.6{\pm}5.5\;and\;0.17{\pm}0.16%$, respectively). Proportions of TUNEL positive cells in NT embryos from FFs and CCs ($6.9{\pm}1.5\;and\;7.4{\pm}1.7%$, respectively) were significantly (P<0.05) higher than in MSCs ($4.8{\pm}1.4%$) and IVF ($2.3{\pm}0.9%$). The results demonstrate that MSCs have a greater potential as donor cells than FFs and CCs in achieving enhanced production of cloned porcine embryos.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.79-86
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• 2001

Objective: Mouse pre-antral follicles require the addition of gonadotropins (Gns) to complete maturation and ovulation of oocyte and antrum formation in vitro. However, we tried examination of in vitro growth of mouse pre-antral follicles in medium without Gns and/or phygiological factors. And also, pre-antral follicles were isolated from ovaries by mechanical method. Our present studies were conducted to evaluate on the growth of follicles and intra-follicular oocytes and antrum formation in vitro of mouse pre-antral follicles in two different media. Methods: Pre-antral follicles ($91{\sim}120{\mu}m$) were isolated mechanically by fine 30G needles not using enzymes from ovaries of 3-6 week-old female ICR mice. Isolated pre-antral follicles were cultured in $20{\mu}l$ droplets of TCM (n=17; follicles: $107.8{\pm}1.58{\mu}m$; oocytes: $57.9{\pm}1.2{\mu}m$) or MEM (n=12; follicles: $109.3{\pm}2.53{\mu}m$; oocytes: $55.4{\pm}1.6{\mu}m$) under mineral oil on the 60 mm culture dish. All experimental media was supplemented with 10% FBS without Gns and/or physiological factors. Pre antral follicles were individually cultured for 8 days. Antram formation and growth of pre-antral follicles and intra-follicular oocytes were evaluated using precalibrated ocular micrometer at X200 magnifications during in vitro culture. Results were analyzed using combination of Student's t-test and Chi-square, and considered statistically significant when p<0.05. Results: Antrum formation had started in two culture media on day 2. On day 8, antrum formation had occurred in 58.3% of pre-antral follicles cultured in DMEM, but only in 23.5% of those cultured in TCM (p=0.0364). Growth of pre-antral follicles and intra-follicular oocytes were observed on day 4 and 8. On day 4, follicular diameter was similar (p=0.1338) in TCM ($119.4{\pm}2.58{\mu}m$) and MEM ($125.4{\pm}4.52{\mu}m$). However, on day 8, diameters of pre-antral follicles cultured in MEM ($168.9{\pm}17.29{\mu}m$) were significantly bigger (p=0.0248) than that in TCM ($126.7{\pm}4.28{\mu}m$). On day 4 and 8, diameters of intra-follicular oocytes were similar in TCM ($67.1{\pm}1.3$ and $72.4{\pm}0.9{\mu}m$) and MEM ($65.2{\pm}1.7$ and $73.3{\pm}1.5{\mu}m$), respectively. Conclusion: We can conform that medium without Gns and/or physiological factors can be used for in vitro antrum formation and growth of pre-antral follicles and intra-follicular oocytes in mouse. In conclusion, MEM supplemented with FBS can be used for growth in vitro of mouse pre-antral follicles isolated mechanically.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.239-244
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• 1998

The purpose of this study is compare IVF cycle outcome in poor responders between clomiphene citrate (CC) stimulated and controlled ovarian hyperstimulation (COH) protocol. A total of 94 patients responding poorly in previous IVF cycles (estradiol<600 pg/ml or less than 3 oocytes retrieved) subsequently underwent either COH (COH group: 122 cycles, 68 patients) or CC-stimulated cycles (CC group: 43 cycles, 26 patients). CC was administered for five consecutive days starting on cycle day 3 at a dose of 100 mg daily. Serial transvaginal ultrasound examination was done from cycle day 8. Urine was collected $3\sim4$ times before hCG injection for the detection of LH surge. The hCG was administered when serum estradiol reached greater than 150 pg/ml and mean follicle diameter>16 mm. In COH group, ovarian stimulation was done using short protocol (GnRH-a/FSH/HMG/hCG). No difference in age or number of transferred embryos was found between CC group and COH group. COH group had significantly (p<0.05) higher mean peak level of $E_2$ ($810{\pm}112$ vs $412{\pm}55$ pg/ml) and greater number of retrieved oocytes ($3.0{\pm}0.2$ vs $2.0{\pm}0.2$) than CC group. CC group had transferred embryos $(1.8{\pm}0.2)$ compared with $(2.1{\pm}0.2)$ in COH group. However, CC group had higher pregnancy rate than COH group per retrieval [26.9% (7/26) vs 6.2% (6/97)], or per transfer [31.8% (7/22) vs 7% (6/86)]. Although cycle cancellation rate in CC group (48.8%) was higher than that of COH group (21.3%), the pregnancy rate per cycle in CC group was still higher (16.3%) than COH group (4.9%). In addition, implantation rate in CC group was 17.5% (7/40), which was significantly (p<0.01) higher than 3.9% (7/180) in COH group. These data suggest that oocyte and embryo quality are lower in COH cycles of poor responders than CC cycles. We suggest that clomiphene citrate stimulated IVF cycle may be more efficient than COH IVF cycle in poor responders in terms of lower costs and higher pregnancy performance.

• Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.293-298
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• 2009

Several studies have reported that nonylphenol (NP) and 2,2', 4,6,6'-pentachlorobiphenyl (PCB104) exhibit estrogenic activity. To investigate the estrogenic potency of NP and PCB104 during oocyte maturation, fully vitellogenic oocytes (0.76 mm diameter in average) of yellow fin goby, Acanthogobius flavimanus, were exposed in vitro to these chemicals at different concentrations (0.1, 1, 10, 100, and 1,000 ng/mL) with the exogenous precursor $17\alpha$-hydroxyprogesterone ($17{\alpha}OHP$) 50 ng/mL in the presence or absence of human chorionic gonadotropin (HCG). The production of testosterone (T), estradiol-$17\beta$ (E2), and $17\alpha,20\beta$-dihydroxy-4-pregnen-3-one ($17\alpha20{\beta}OHP$) in response to NP or PCB104 were measured by radioimmunoassay. Steroid levels were also expressed as E2/T and E2/$17\alpha20{\beta}OHP$ ratios. In the absence of HCG, no significant differences in either NP or PCB104 treatment groups were observed. In the presence of HCG, NP treatment did not show significant differences in the production of T, E2, and $17\alpha20{\beta}OHP$ at any concentrations tested, but E2/T ratios were decreased at concentrations of 0.1, 1, 10, and 1,000 ng/mL compared with the control group. PCB104 decreased E2 production at concentrations of 0.1, 10, and 1000 ng/mL, but did not show significant differences in the production of T and $17\alpha20{\beta}OHP$ at any concentration tested. While E2/T ratios were decreased at PCB104 concentrations of 0.1, 1, 10, and 1,000 ng/mL, E2/$17\alpha20{\beta}OHP$ ratios were also decreased at 0.1, 10, and 1,000 ng/mL compared with the control. Results indicate that both NP and PCB104 appeared to have antiestrogenic effects during this phase.

• Journal of Life Science
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• v.13 no.5
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• pp.732-739
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• 2003

The purpose of this study was to determine if the addition of spermatozoa into the culture medium could influence the nuclear maturation of denuded porcine germinal vesicle (GV) oocytes in vitro. Cumulus-oocyte complexes were collected from follicles of 3 to 5 mm in diameter, The cumulus and corona cells were removed from oocytes. Porcine denuded oocytes were cultured in tissue culture medium containing spermatozoa. After 48 h culture, oocytes were examined for the evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II (M II). The proportion of oocytes reaching M II stage was significantly (P<0.01) increased in the oocytes cultured in media containing spermatozoa compared to those in media without spermatozoa $(31.9\pm1.8%\; vs\; 14.9\pm1.0%)$.No differences in the rates of M II were observed among the different period of spermatozoa exposure nor among the spermatozoa from different species. The proportion of oocytes reaching M II stage was significantly different between high and low concentrations of spermatozoa. The present study suggests that mammalian spermatozoa contain a substance(s) that improves nuclear maturation in vitro of GV oocytes. Enhancing effect of spermatozoa for oocytes maturation in vitro is a highly dose-dependent.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.22 no.5
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• pp.342-350
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• 1989

Maturity and spawning of Crangon affinis were studied based on the histological observation. The samples were monthly collected in Nakdong estuary from June, 1988 to May, 1989. The gonad lies on the dorsal side of the thorax. The cavity in which it lies is located below heart and above hepatopancreas. Anterior part of ovary is fused roundly, and the posterior part shows a pair of tubule-like structure. Testis is bilaterally symmetrical; the anterior part shows convoluted tubule, and the posterior part consists of a pair of tubule. Seminal vesicle is connected and opened to the base of the fifth pereiopod. Gametogenesis of ovum and spermatozoon is being repeated in short period without seasonality, and it is formed simultaneously in the whole parts of germinal epithelium. Diameter of a ripe oocyte is ca. $430{\mu}m$. Spermatozoa is oval with distinct nucleoplasm. Reproductive cycle is performed in ca. 40 days. Crangon affinis spawns all year round except November. Incubation period of brooded eggs took $12\~14$ days before hatching in the aquarium at $21^{\circ}C\~24^{\circ}C\;and\;33\%_{\circ}$.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.83-88
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• 2000

This study was conducted to determine the effect of repeated ultrasound-guided transvaginal retrieval of oocytes recovery, estrous cycle and ovarian adhesion in Korean native, Hanwoo heifers. Heifers were at unknown stages of the estrous cycle at the start of experiments in which all follicles $\geq$6mm in diameter were ablated. The results obtained in this study were as follows; Follicle developing number and oocytes collected number were no effected to repeated OPU to nine session, 4 e.a range oocytes collected to repeated OPU session. Oocytes were observated follicles were 8.7$\pm$4.2 e.a, collected oocytes were 4.1$\pm$3.4 e.a to two times collected per week and observated follicles was 10.2$\pm$6.1 e.a, collected oocytes were 4.3$\pm$2.9 e.a to one times collected per week, but no difference significantly(P＜0.05). Ovaries adhesive percentage to repeated OPU was eight ovaries adhisived(20%) of forty ovaries, three ovaries(7.5%) to 1~3 times oocytes collected, four ovaries(l0%) to 4~6 times, one ovaries(2.5%) to 7~9 times oocytes collected session. To repeated OPU effection, ovaries adhisive heifers were long estrous cycle(＞25 day) to 7 heads(87.5%) of 8 head, non-adhesive ovaries heifers were 5 heads(41.7%) were long estrous cycle to repeated OPU 12 heads. Although, now unknown about the dynamics of follicles wave and about functional changes to repeated OPU ovaries, more question about ovaries adhesive cause remain.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.233-240
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• 1994

This study was conducted to compare the endocrine milieu, and pregnancy rates in In Vitro Fertilization and Embryo Transfer(IVF-ET) program employing combined with gonadotropin releasing hormone agonist(GnRH-a) and pergonal(LH 75lU+FSH 75lU) when either human chorionic gonadotropin(HCG) or progesterone were used for luteal phase support. A total number of 40 IVF-ET treatment cycles were prospectively studied. Ovarian hyperstimulation method was modified ultrashort protocol using GnRH-a. All patients started Decapeptyl at menstrual cycle day # 2, and HMG was started at # 3 days. When leading follicle was ${\geqq}$18mm or at least two follicles were ${\geqq}$14mm in diameter, HCG 10000lU intramuscularly was injected. After 36 hours HCG administration, oocytes were retrieved as usual guided by transvaginal ultrasound. Embryo were transfered 36-48 hours later. The patient's cycles were prospectively randomized to receive HCG(20cycles) or Progesterone (20cycles) for luteal support. The progesterone group received 25mg 1M starting from the day of ET. The HCG group received 1500IU 1M. on days 0, +2, +5 after ET. Estadiol($E_2$) and Progesterone($P_4$) were measured on the day of oocyte aspiration, ET day, and every 6 days thereafter. Results were follows as; 1. Estradiol, progesterone and LH levels on the day of HCG trigger, retrieved oocytes and number of transfered embryo were not significantly different in both groups. 2. On the day of aspiration and embryo transfered day, $E_2$, $P_4$ level were significantly higher in progesterone group than HCG group(p<0.01). 3. $E_2$, $P_4$ level on 6 days after ET were significantly higher in progesterone group than HCG group(p<0.01). But, $P_4/E_2$ ratio was not different in both groups. 4. $E_2$, $P_4$ level 12 days after ET were decreased abruptly in both groups and higher hormonal level appeared in HCG group(P<0.01). 5. The total pregnancy rate in the HCG group was 40% (8/20) and in the progesterone group 15%(3/20). 6. Comparing the pregnant and nonpregnant cases progesterone group was not different the hormonal status. In HCG group, pregnant cases appeared in higher $P_4$, $P_4/E_2$ ratio than nonpregnanct cases(P<0.01).