• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2001년도 정기총회 및 봄 학술발표회 초록집
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• pp.238-239
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• 2001

The egg-yolk precursor vitellogenin(VTG) is secreted by the liver of female and male fish, in response to estrogenic compound. Carp(Cryprinus carpio) vitellogenin of one major protein is 160kDa, two minor proteins is 110kDa and 170kDa. We were induced vitellogenin by inject of 17-estradiol and purified in a two step procedure by separating it from plasma protein precipitated by 35% saturated ${(NH_4)}_2SO_4$ and then from the remainder by Mono-Q chromatography. We found major carp(Cyprinus carpio) vitellogenin band at 160kDa. Effect of different concentration of oestrogen on vitellogenin synthesis in carp(Cyprinus carpio) exposed for 4 weeks. Show differential effects on vitellogenin synthesis 7 days after treatment. Plasma vitellogenin was measured 3 times for each by an enzyme linked immunosorbent assay(ELISA). ELISA was developed for the detection of the egg yolk precursor vitellogenin in plasma of carp(Cyprinus carpio). The ELISAS performance was optimized and characterized.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.355-361
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• 2011

Effects of aluminum (Al) on plasma vitellogenin (VTG), alkaline-labile phosphorus (ALPP), calcium (Ca), glutamate pyruvate transaminase (GPT), the hepatosomatic index (HSI), and hepatic Al concentration were examined in estradiol-$17{\beta}$ ($E_2$)-administered immature rockfish Sebastes schlegeli. Fish were injected intraperitoneally with $E_2$ (5 mg/kg body weight [BW]) and/or Al (0, 0.1, 1, 5, and 10 mg/kg BW) and plasma and liver samples were extracted 7 days later. After sodium dodecyl sulfate polyacrylamide gel electrophoresis, the relative amount of VTG was determined by integrated optical density. VTG accounted for 23.6% of the total proteins in the control group, but this value decreased with increasing Al administration. Al reduced the concentrations of ALPP and Ca in a concentration-dependent manner and significant reduction occurred at Al concentrations greater than 5 mg/kg. The concentration of GPT increased in a concentration-dependent manner in all Al-administered rockfish. The concentrations of Al in the liver also increased, and HSI was decreased, in a concentration-dependent manner. These results suggest that Al inhibits $E_2$-induced VTG production by being toxic to hepatocytes in marine fish.

• Asian-Australasian Journal of Animal Sciences
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• 제24권9호
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• pp.1204-1210
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• 2011

Objective of this study was to examine the effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor on steroid synthesis, growth and apoptosis of buffalo granulosa cells (GCs) in vitro. Follicular fluid of antral follicles (3-5 mm diameter) was aspirated and GCs were cultured in 0 (control), $10^{-3}$, $10^{-5}$, $10^{-7}$, $10^{-9}\;M$ of SNP for 48 h. To evaluate whether this effect was reversible, GCs were cultured in presence of $10^{-5}\;M$ SNP+1.0 mM $N^{\omega}$-nitro-L-arginine methyl ester (L-NAME) a NO synthase (NOS) inhibitor or hemoglobin (Hb, $1.0{\mu}g$) as NO scavenger. Nitrate/nitrite concentration was evaluated by Griess method, progesterone and estradiol concentrations by RIA and apoptosis by TUNEL assay. SNP ($10^{-3}$, $10^{-5}$, $10^{-7}\;M$) significantly (p<0.05) inhibited estradiol and progesterone synthesis, growth, disorganized GCs aggregates and induced apoptosis in a dose dependent manner. However, $10^{-9}\;M$ SNP induced the progesterone synthesis and stimulated GCs to develop into a uniform monolayer. Combination of SNP $10^{-5}$ M+L-NAME strengthened the inhibitory effect while, SNP+Hb together reversed these inhibitory effects. In conclusion, SNP at greater concentrations ($10^{-3}$, $10^{-5}$ and $10^{-7}\;M$) has a cytotoxic effect and it may lead to cell death whereas, at a lower concentration ($10^{-9}\;M$) induced progesterone synthesis and growth of GCs. These findings have important implications that NOS derived NO are involved at physiological level during growth and development of buffalo GCs which regulates the steroidogenesis, growth and apoptosis.

• 한국가축번식학회지
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• 제24권2호
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• pp.163-170
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• 2000

본 연구는 rBST, vitamin E 및 selenium (Se)의 투여가 한우 종모우의 혈액성분 및 호르몬의 농도변화에 미치는 영향을 검토하였다. 1. rBST 투여구, Vit. E 투여구 및 Se 투여가 한우 종모우의 혈액성분에 미치는 효과를 조사한 결과, 혈중 albumin의 함량은 Se 투여구가 대조구, rBST 투여구 및 Vit. E 투여구보다 통계적으로 유의하게 높은 함량을 나타났으며 (P＜0.05), 혼합투여구는 여타구 (대조구, rBST 투여구 및 vitamin E 투여구)보다 통계적으로 유의하게 높게 나타났다 (P＜0.05). 혈중 BUN (Blood Urea Nitrogen)과 creatinine의 함량은 rBST 투여구가 여타구보다 통계적으로 유의하게 높게 나타났다 (P＜0.05). 혈중 total protein 함량은 rBST 투여구, Se 투여구 및 혼합투여구가 대조구보다 통계적으로 유의하게 높은 함량을 나타냈다 (P＜0.05). 혈중 calcium, cholesterol, glucose, inorganic phosphoros 의 함량은 투여구간에 차이가 없었다 (P＞0.05). 2. rBST, Vit. E 및 Se 투여가 한우 종모우의 혈중내 호르몬의 변화에 미치는 효과를 조사한 결과, 혈중 estradiol의 농도는 Se 투여구와 혼합투여구가 여타구 (대조구, rBST 투여구 및 Vit. E 투여구 )보다 다소 높게 나타났지만 유의차는 인정되지 않았으며 (P＞0.05), 혈중 testeserone의 농도는 투여구간에 커다란 차이는 없었다 (P＞0.05).

• 한국양식학회지
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• 제11권2호
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• pp.279-282
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• 1998

Effects of high concentration of intracellular calcium on estradiol-induced vitellogenin(VTG) induction were examined using ouabain in Primary hepatocyte culture in the rainbow trout Oncorhynchus mykiss. Ouabain increases cytosolic free calcium as a result of inhibition of $Na^+ - Ca^{2+}$ exchanger. Ouabain markedly reduced VTG production to the control level, despite of calcium concentrations in the incubatin medium. Therefore, ouabain would reduce VTG production not by increasing intracellular calcium bt directly by inhibiting $Na^+ - K^+$ ATPase.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.164-164
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• 2002

The estrogenic activity of 47 plant extracts was assessed by reporter gene assay using MCF-7 breast cancer cell lines stably transfected with luciferase reporter gene. The estrogenic activity of food extracts was expressed as 17${\beta}$-estradiol(E2) equivalent concentration(EEQ), the concentration of E2 that resulted in the same relative luciferase unit(RLU) of the food extract of 0.2mg/$m\ell$.(omitted)

• 한국양식학회지
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• 제11권3호
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• pp.311-317
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• 1998

Estradiol-17${\beta}$($E_2$)에 의한 Vitellogenin (VTG) 함성에 미치는 Cu 및 Zn의 영향을 무지개송어의 배양 간세포를 이용하여 조사하였다. 간세포는 2일간 배양 한 후, E하(2) ($2{\times}10^6$ M)와 동시에 Cu ($2{\times}10^{-5}$~$10^{-4}$M) 또는 Zn ($10^{-5}$~$10^{-3}$M)을 배양액에 첨가하여 5일간 배양하였다. VTG 합성률은 총 단백질에 대한 VTG의 배율로 나타내었다. CU 및 Zn의 첨가는 배양 간세포의 생존율에는 영향을 미치지 않았다. 그러나, Cu의 첨가에 사용된 모든 농도에서, Zn의 첨가에는 농도 의존적으로 감소하여(10^{-3}$M에서 유의하게 감소하였다. 이러한 감소는, Zn의 제거시에는 회복되었으나, Cu에서는 회복되지 않았다. 그리고, Cu (10^{-4}$M의 첨가 시에 Ca (1.8 mM) 농도를 2.5 및 5.0 mM로 증가시켜도 Cu의 작용을 저해하지 못하였다. 이러한 결과로 보아, Cu 및 Zn은 간세포에서 합성되는 다른 단백질 보다 VTG 합성에 더 깊이 관여하는 것으로 추정된다.

• 한국가축번식학회지
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• 제14권3호
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• pp.175-182
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• 1990

Behavioural estrus and short estrous cycles were observed and serum concentrations of estradiol-17$\beta$(E2) before and after of estrous were measured following superovulation treatments in 30 pluriparous Korean native goats. The goats were divided into 2 groups. Fifteen goats were injected IM with 1,000IU PMSG on Day 12 of the estrous cycle followed by 10mg PGF2$\alpha$ 48h later(P4＋PMSG), and the other 15 goats were injected IM with 10mg progesterone(P4)in oil once daily for 10d beginning at any days of estrous cycle followed by 1,000IU PMSG and 10mg PGF2$\alpha$ at the 8th day of progesterone treatment(P4＋PMSG group). After injection of PGF2$\alpha$, onset of standing estrus occurred in 12 of 15 goats(80.0%) at 50.0$\pm$7.7h and in 11 of 15 goats(73.3%) at 135.6$\pm$10.1h in PMSG and group and P4＋PMSG group, respectively. The mean interval from PGF2$\alpha$ injection to first estrus was significantly(P<0.01) earlier in PMSG group than in P4＋PMSG group. This result indicate that the delayed infusion of P4 in P4＋PMSG group caused the later exhibition of their estrous behaviors. However, duration fo frist estrus(31.5$\pm$2.6h vs 26.2$\pm$2.3h), length of estrous cycle(14.1$\pm$3.3d vs 16.6$\pm$3.8d) and percentage of short estrous cycle(50.0% vs 45.5%) were not different between PMSG and P4＋PMSG group. The mean concentration of serum E2 in 4 goats showing normal estrous cycle in P4＋PMSG group(PP-NEC) was higher than in 6 goats showing normal(P-NEC) or in 6 goats showing short estrous cycle(P-SEC) in PMSG group. The peak level of serum E2 was observed at the time of onset of standing estrus in PP-NEC(67.6pg/ml), 6h earlier in P-NEC(53.1pg/ml) and 6h later in P-SEC(52.3pg/ml) than the onset of standing estrus. The profiles of serum concentration of E2 during the period of peri-estrus was similar in the goats of PMSG or P4＋PMSG and also in the goats showing the subsequent estrous cycle of normal or short length.

• 한국수정란이식학회지
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• 제12권1호
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• pp.27-36
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• 1997

The studies were carried out to investigate the effects of co-culture with cumulus cells and oviduct epithelial cells on the in vitro fertilization and cleavage rate of bovine follicular cocytes and to determine the optimum thawing temperature and equilibration time on in vitro developmental rate of frozen bovine embryos. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TGM-199 medium containing 10 IU /ml의 PM SG, 10 IU /ml의 hCG, ip g/ml의 $\beta$-estradiol and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquld nitrogen and thawed in 3$0^{\circ}C$ water. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are sunanarized as followes :1. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with cumulus cells in TCM499 medium were 75.0~76.8% and 17.3~27.6%, respect-ively. And in-vitro fertilization rates of cumulus-enclosed oocytes(55.4%)were significantly(p<0.05) higher than cumulus-denuded oocytes (23.1%). 2. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with l$\times$ l04cells /ml, 1 x l06cells /ml, lx l08cells /ml and 1 x l015cells /ml oviduct epithelial cells in TCM-199 medium were 74.5~77.8% and 15.7~21.20 respectively.3. The in-vitro fertilization and in vitro developmental rates of bovine oocytes cocultured in '1CM-199 media containing PMSG, hCG, PMSG+hCG. PMSG+$\beta$-estradiol, hCG+$\beta$-estradiol 0 to 40 hrs after insemination were 74.0~77.4% and l8.9~23.l%, re-spectiv ely.4.The survival rates of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol,DMSO and propanediol were 23.5~31.4% and 20.6~34.l%, respectively. 5. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$.6. The equilibration time on the survival rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20min.). (Key words : bovine embryos, co-culture, freezing, in vitro development)

• 생명과학회지
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• 제19권12호
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• pp.1764-1768
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• 2009

본 연구는 소의 과립막세포 배양상층액에서 스테로이드 호르몬의 농도와 생쥐 배의 체외 발달에 미치는 효과를 검토하기 위하여 수행되었다. 소의 성숙난포(직경 6~15 mm)와 미성숙 난포(직경 2~5 mm)로부터 과립막세포를 각각 분리하여 15% FCS가 첨가된 Ham's F-10 배양약에서 16일간 배양하였다. 과립막세포들은 쉽게 단층배엽을 형성 하였으며, 배양과정 중 비슷한 성장패턴을 나타내었다. 또한 성숙 과립막세포와 미성숙 과립막세포 간에 형태적인 차이가 없었다. 과립막세포 배양 중 progesterone과 estradiol 생산이 활발하게 이루어졌으며, progesterone은 배양후기에, estradiol은 배양초기에 분비량이 높았다. 성숙 과립막세포와 미성숙 과립막세포에서 내분비 양상은 비슷하였다. 생쥐배가 상실배, 배반포 및 부화배반포 단계로 체외발달된 비율은 Ham'F-10배양액(86.7%, 41.7%, 13.3%)에 비하여 성숙 과립막세포 배양상층액(92.7%, 78.1%, 34.5%)과 미성숙 과립막세포 배양상층액(96.4%, 78.5%, 26.8%)에서 유의적으로 높았다(p<0.05). 그러나 배의 발달에 대하여 성숙 과립막세포배양상층액과 미성숙 과립막세포 배양상층액 간에는 차이가 없었다.