• The Korean Journal of Zoology
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• v.30 no.1
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• pp.1-9
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• 1987

Cyclic AMP (cAMP) was known to play a key role in the regulation of cumulus expansion and oocyte maturation of mammalian cumulus-oocyte complexes (COC's) in vivo and in vitro. The present experiments were conducted to know how intracellular level of cAMP in these cells is controlled. Intracellular cAMP level was modulated by culturing mouse CGC's with an adenylate cyclase stimulator, forskolin, a phosphodiesterase inhibitor, 3-isobutyl-1-methyixanthine (IBMX), human chorionic gonadotrophin (HCG), or follicle stimulating hormone (FSH). The rate of cumulus expansion and germinal vesicle break-down (GVBD) was checked after culture and used as a biological end point. Forskolin in the medium began to stimulate the expansion of the complexes at 1 nM and induced maximum expansion (80~90%) at 0 1~10 $\mu$M. The expansion rate was reduced to 60% when forskolin concentration was increased to 100 $\mu$M. Oocyte GVBD occurred normally (75~82%) in the presence of 10 $\mu$M of forskolin, but partial suppression was appeared at 100 pM of the drug (40%). IBMX also stimulated the expansion from the concentration of 0.01 pM and induced full expansion (81~89%) between the concentration of 1-1000 $\mu$M. Meiotic resumption was occurred normally under 10 $\mu$M of IBMX, but suppressed drastically from the concentration of 100 $\mu$M. The minimum exposing time to hormone or drugs required to trigger cumulus expansion was two minutes with HCG, 15~30 minutes with FSH and fors kolin, and two hours with IBMX. The data presented here seemed to imply that intracellular cAMP level in cumulus cells is regulated by both adenylate cyclase and phosphodiesterase and cumulus expansion is induced by a peak of cAMP while meiotic arrest is maintained by continuous presence of cAMP.

• 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.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.79-86
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• 1997

This study was designed to investigate the effects of superovulation on the growing and mature follicles following gonadotrophin treatments in mature rat by immunohistochemical methods. Eighteen mature rats (Sprague-Duwely, 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone(FSH) / rat, and third PMS and HCG-treated group was injected intramuscularly with 20~25IU of pregnant mare serum(PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotropin(HCG) / rat. Half the number of rats were administrated intraperitoneally with bromodeoxyuridine(Brdur, 0.2mg/gm BW once) at 2 hours before exanguination and the remainder of rats were sacrified without Brdur administration. The investigation by immunohistochemical methods using paraffin sections of ovaries was performed by using anti-Brdur antibody and PCNA(proliferating cell nuclear antigen) antibody for labeling proliferating cells in follicles. In immunohistochemical findings, follicles squeezed by peripheral corpus luteum or follicles large follicles with loosly and irregularly distributed granulosa cells and although with compacted granulosa cells, middle follicles with dilated round or oval follicular antrum were confirmed as atretic follicles. The proportions of atretic follicles in control group were 29.8%, 21.7% and 14.2% respectivley at large, middle and small follicles and mean proportions of these all 3 grade follicles were 26.7%. The proportions of atretic follicles in FSH-treated group were 35.4%, 24.9% and 10.4% respectively at large, middle and small follicles and mean proportions of these all 3 grade follicles were 28.1%. The proportions of atretic follicles in PMS and HCG-treated group were 44.7%, 24.0% and 12.7% respectively at large, middle and small follicles, and mean proportions of these all 3 grade follicles were 29.7%. The above findings reveal that the group with higher proportion of atretic follicles were ordered as large, middle and small follicles in size, and these proportions were increased in hormone treated two groups with more number of more growing and mature follicles when compared with control group.

• Journal of Aquaculture
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• v.10 no.4
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• pp.463-472
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• 1997

Floating and hatching rates, size and survival activity index(SAI) of spawned eggs of red spotted grouper, Epinephelus akaara were studied to understand egg qualities during the spawning periods in 1992, 1993 and 1995. The fish spawned from mid-July to mid-August when temperature ranged 23~$28^\circC$. Floating, fertilization and hatching rates were higher in the eggs from early to middle than those from late during the spawning period. SAI ranged 1.8~9.6, and it was closely related to survival rate until mouth operning time (SMOT). Egg diameter decreased gradually with the time of spawning, while diameter of oil globule was constant during the spawning periods. Induced spawnings were carried out with one injection of HCG over 5, 000 IU/kg body weight. Floating, fertilization and hatching rates of the induced spawning eggs were higher than those of untreated ones. Thus, induced spawning method with HCG was recommended for collecting fertilized eggs in bulk. Optimal time for the collection of high quality eggs was supposed to be from early to middle during the spawning periods. Considering that egg qualities were vulnerable within the period, hatchability, incidence of malformed larvae, SMOT and SAI were factors to be considered together to get good quality of eggs.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.333-340
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• 2003

Objective: This study was performed to examine the maturation and the development to the blastocyst stage of immature oocytes collected from patients with high risk of ovarian hyperstimulation syndrome (OHSS). Materials and Methods: Cumulus-oocyte complexes (COCs) were collected following only HCGpriming for non stimulated IVF-ET cycles of the patients. At the time of oocyte collection, COCs were classified into three groups in accordance with their appearance (Group I: oocytes with dispersed cumulus cells; Group II: oocytes with compacted cumulus cells; Group III: oocytes with sparse cumulus cells). The in vitro maturation and blastocyst development rates of the COCs were compared among these groups. From August 2001 to June 2002, 48 IVM/IVF-ET cycles from 42 patients (mean age: $32.4{\pm}3.8$ years) were performed. To prevent the occurrence of OHSS, the patients were primed with 10, 000 IU HCG alone 36 h before oocyte collection without gonadotropin stimulation. Oocytes were aspirated on cycle days from 7 to 13. The normal COCs were classified into three groups according to their appearance. The aspirated immature oocytes were cultured in YS maturation medium containing 30% (v/v) human follicular fluid (HFF), 1 IU/ml FSH, 10 IU/ml HCG and 10 ng/ml rhEGF. Fertilization was induced by intracytoplasmic sperm injection (ICSI). All zygotes were co-cultured with cumulus cells in $10{\mu}l$ YS medium containing 10% HFF until day 7 after oocyte collection. Blastocyst transfer was performed on day 5 after ICSI. Results: Th e mean number of oocytes cultured in the IVM/IVF cycles was $24.7{\pm}10.6$. Of 1185 COCs, those assigned to Group I, II and III were 470 (39.7%), 414 (35.0%) and 301 (25.4%), respectively. The maturation rate (94.5%, 444/470, p<0.05) in Group I was significantly higher than those of Group II (62.8%, 260/414) and Group III (73.1%, 220/301). Especially, 30.9% of COCs in Group I (145/470) was matured on the day of oocyte aspiration. There were no differences in the rates of fertilization and cleavage among the three groups. The development rate to the blastocyst stage in Group I (54.6%, 206/377, p<0.05) was also significantly higher than those in Group II (33.0%, 68/206) and Group III (30.1%, 52/173). Twenty-four clinical pregnancies (50.0%) was obtained and 22 pregnancies (45.8%) are ongoing. Implantation rate in the present study was 24.6%. Conclusion: These results suggest that there is a positive correlation between the appearance of COCs and the developmental competence of the immature oocytes in non stimulated IVM/IVF cycles.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.219-219
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• 2004

Human erythropoietin (hEPO) 유전자가 도입된 형질전환돼지는 수태와 임신율을 일반돼지와 비교를 해 보았을 때 매우 낮은 경향을 보이는 것으로 나타났으며, 이러한 경향의 원인을 구명하기 위하여 인위적인 발정동기화 및 호르몬을 이용한 과배란 유기시 배란과 수정(수태)시 성호르몬 변화를 비교하고자 실시하였다. 먼저 발정동기화를 위하여 PG 600 (PMSG 400 IU, HCG 200IU)을 주사한 후 10일 후에 Regumate porcine을 6일간 급여한 후에 PMSG 주사하고 2일 후에 hCG 주사한 다음 인공수정을 실시하였다. (중략)

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.282-283
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• 2000

The egg development of various fishes is studied by various methods including electron microscopy(Matsuyama et al, 1991) and hormone treatment(Schoonbee et al. 1980). In addition to attempt to induce final maturation and spawning, the present experiments were conducted to investigate the influence of hCG upon reproductive cycles in catfish(Silurus asotus) with developing gonads and to stimulate the reproductive system during the rapid phase of gonadal development. (omitted)

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.276-277
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• 2000

어류의 성숙 및 배란을 유도하기 위해 어류 및 개구리의 뇌하수체 추출물, 인간의 태반성생식소자극호르몬(HCG), 어류의 생식소자극호르몬(GTH), 포유류의 황체형성호르몬방출호르몬(LHRH) 그리고 이들의 유사체(analogue)들이 주로 이용되고 있다. 특히 시상하부의 신경분비 세포에서 분비되는 LHRH를 이용한 어류의 배란 및 산란에 대한 연구는 여러 어종에 대해서 시도되어 왔다. (중략)

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.331-333
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• 1994

A 23-year-old twin brother visited our hospital for the evaluation of anosmia and delayed secondary sexual maturation. They show eunuchoid apperance, gynecomastia, micropenis and scanty pubic hair. On hormonal study, they show findings of hypogonadotropic hypogonadism. So they were treated with HCG for 3 months and thereafter with testosterone. Herein we report a case of Kallman's syndrome in twin brother.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.51-51
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• 2003

대농갱이는 체형이 동자개와 비슷하나 지금까지 인공부화는 물론 산란습성에 관해서도 잘 알려져 있지 않은 미개발 어종으로 양식품종의 다양화 및 경쟁력 있는 새로운 품종 개발을 위하여 인공종묘생산 시험을 시도하였다. 본 연구에 사용한 대농갱이 친어는 금강 중류역에서 서식하고 있는 자연산 친어로 암컷 210마리, 수컷 148마리를 사용하였다. 호르몬주사에 의한 인공 배란효과 조사를 위해 암컷 한 마리당 HCG 1,000 IU 주사군(75마리)과 2,000 IU 주사군(135마리)으로 나누어 배란율을 조사하였으며, 수컷은 주사하지 않고 정소를 적출하여 막자사발에 갈아서 등조액과 혼합하여 사용했다. 수정 방법은 난을 착출하면서 정자 등조액을 동시에 흘리는 방법, 난을 그릇에 받은 후 정자 등조액을 혼합 수정하는 방법, 채란용기에 정자 등조액을 미리 넣고 암컷의 난을 짜 넣어 수정시키는 방법 등 3가지 방법을 사용하였다. 부화된 자어는 초기먹이로 알테미아(3일 급이), 실지렁이(6일 급이), 배합사료 순으로 급이하였으며 수온 25~28$^{\circ}C$에 유지관리하면서 성장도 시험을 실시하였다. 호르몬 주사에 의한 배란효과는 HCG 2,000 IU 주사군(채란율 88.9%)이 1,000 IU 주사군(37.3%)에 비해 월등히 높았으며 난의 성숙상태도 좋았다. 수정방법에 있어서는 채란과 동시에 정자 등조액을 흘려 수정시키는 방법과 채란용기에 미리 정자 등조액을 넣고 암컷의 난을 한 마리씩 짜서 수정한 후 바로 부화지로 이동하여 고르게 뿌려 준 경우가 수정율 및 부화율이 좋았다. 부화수온은 23$\pm$1$^{\circ}C$에서 96시간, 26$\pm$1$^{\circ}C$에서는 72시간만에 부화되었으며 부화여건이 좋으면 단시간에 부화할 수 있도록 하는 것이 수생균 발생을 억제할 수 있어서 효과적이었다. 자치어의 성장도 실험에서는 사육수온 25~28$^{\circ}C$에서 직선식으로 나타낼 수 있어서 평균체장은 Y=0.9943X+0.7867, 평균체중은 Y=0.8800-0.6800로 각각 표시되었다.