• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2003년도 International Symposium on Clean Environment
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• pp.159-164
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• 2003

Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r＝-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

• 한국발생생물학회지:발생과생식
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• 제9권2호
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• pp.135-142
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• 2005

무척추동물에서 척추동물에 이르기까지 대부분의 난생동물들의 난황 단백질의 전구체를 vitellogenin(VTG)이라 한다. 난생 척추동물에서 VTG는 간에서 합성되어 혈액을 통해 난세포로 전이된다. 암컷 어류는 정상적인 생식주기에서 난황단백전구체 형성이 시작되면 혈중 농도는 급격히 증가하게 된다. 수컷도 VTG의 유전자를 가지고 있기 때문에 낮은 수준의 내인성 에스트로겐으로 아주 적은 양의 단백질이 있을 수 있다. 그렇지만 수컷에 외인성에스트로겐 유사물질에 노출되면 그 양은 증가하게 된다. 따라서 어류에서 VTG는 내분비계 장애물질 효과를 조사하는 유용한 생물학적 추적자로 사용할 수 있다. 이 연구에서는 에스트로겐 유사물질에 오염된 지역에 서식하는 망둑어류의 혈중 VTG의 수준을 정량할 수 있는 방법을 제공하고자 한다. 이러한 목적을 위해 꾹저구(Chaenogobius annularis)의 VTG를 분리하고, 이에 대한 단클론 항체와 다클론 항체를 제작하여 혈중 VTG 수준을 정량할 수 있는 sandwich 경합 ELISA system을 만들었다. 난황 단백질에 대한 단클론 항체와 다클론 항체를 사용한 ELISA system에 대한 타당성 검토를 하였다. 순차적으로 희석한 암 컷의 혈청에 대한 흡광도 곡선은 VTG 표준농도의 곡선과 평행하였으나 수컷의 혈청에 대한 흡광도 곡선은 평행하지 않았다. 이 VTG에 대한 sandwich ELISA system으로 꾹저구(C. annularis)의 혈중 VTG의 수준을 정량뿐만 아니라 문절망둑(Acanthogobius flaviman)과 풀망둑(A. hasta)의 혈중 VTG 수준을 정량할 수 있다.

• Fisheries and Aquatic Sciences
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• 제7권3호
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• pp.99-108
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• 2004

Vitellogenin (VTG) was purified from serum of $estradiol-l7{\beta}-treated rockfish$(Sebastes schlegeli) by precipitation with $EDTA-Mg^{2+}$ and ammonium sulfate and two step chromatography (anion exchange chromatography and gel permeation chromatography) was performed on FPLC system. Rockfish VTG (rfVTG) was characterized and its properties were determined. The monomers have apparent, molecular mass of about 188 kDa as indicated by SDS-PAGE. Amino acid composition analysis of rfVTG was similar to VTG from other oviparous teleosts. Cysteine and lysine were present at relatively high level. Leucine was present at relatively lower level than in other species. The N-terminal amino acid sequence was evaluated to identify rfVTG. Western blot analysis using an antibody against the purified VTG showed that the antibody reacted with both plasma of $estradiol-l7{\beta}-treated rockfish$ treated male and purified VTG, whereas there was no reaction with male serum of the control. An ELISA was developed using monoclonal and polyclonal antibodies against rfVTG. The assay range was 3.2 ng/mL and 1,000 ng/mL and the value of the intra and inter assay variations were within $9.7{\%}$ and $11.2{\%}$, respectively. Recovery rate was $96.8{\%}$. The sandwich ELISA could be useful for the detection of VTG and could be good for screening of estrogenic compounds.

• 한국양식학회지
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• 제9권1호
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• pp.83-92
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• 1996

농어목 어류, C. diagramma 암컷의 vitellogenin과 난황단백의 면역화학적 특성을 서로 비교하였다. 면역화학적 방법에 의해 C. diagramma 암컷의 혈청에서 vitellogenin 1 (VTG1)과 vitellogenin 2 (VTG2)를 동정하였으며, sephacryl S-300 gel 여과법에 의하여 측정된 분자량은 각각 560,000과 410,000이었다. 암컷의 난소 추출액에서는 두 가지 형태의 난황단백인 E2와 E3가 분리되었으며, 이들의 분자량은 각각 410,000과 170,000이었다. $17\beta$-estradiol ($E_2$)을 주사한 수컷의 혈청과 간조직을 abidine-biotin complex (ABC) 방법으로 분석한 결과, vitellogenin과 유사한 물질이 검출되었다. 이상의 결과로 부터, 외인성 $E_2$ 에 의해 간에서 합성된 vitellogenin은 혈액 속으로 방출된 후 난소로 이동되는 것으로 추정되었다. 암컷의 $E_2$, 혈중농도는 산란기인 6월에 최고치를 나타내었고, vitellogenin 상대농도도 $5\~6$월에 최고치를 나타내었다. $E_2$와 vitellogenin의 변화는 본 종의 난소성숙과 밀접한 관계가 있으며, 난황형성에 있어서도 중요한 역할을 하고 있음을 알 수 있었다. 암컷 혈청 VTG2와 난소추출액 E2의 분자량이 서로 같고, vitellogenin이 간에서 합성되어 난소내로 이동하는 점으로 보아, 난소내 난황단백 성분의 전구물질은 혈청내의 vitellogenin인 것으로 추정되었다.

• Fisheries and Aquatic Sciences
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• 제4권4호
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• pp.186-191
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• 2001

Vitellogenin (VTG) and VTG mRNA induction by $estradiol-17\beta\;(E_2)$ were examined in the primary cultures of hepatocyte in the rainbow trout. Hepatocytes were precultured for 2 days, then $E_2$ was added and cultured for another 5 days. Media and hepatocytes were then analyzed by electrophoresis and Northern blotting for VTG and VTG mRNA, respectively. The hepatocytes were formed a few aggregates within 5 days without further spreading to a monolayer. Cell viability and high DNA content were maintained during the incubation. The hepatocyte culture with E2 induced a weak VTG band at a molecular weight of 175kDa on Day 2 after $E_2$ addition. The relative amount of VTG was expressed in percentage of total protein concentrations. VTG was gradually increased as $1.9\%$ on Day 2, $6.3\%$ on Day 4 and $7.3\%$ on Day 5. VTG mRNA band was detected at about 6.6 kb in the culture with $E_2$ at day 1 of culture. The level of VTG mRNA expression linearly increased with time until Day 5 (r=0.97).

• 한국정보처리학회:학술대회논문집
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• 한국정보처리학회 2001년도 추계학술발표논문집 (상)
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• pp.535-538
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• 2001

Since 1990s, many literatures have shown that connectionist models, such as back propagation, recurrent network, and RBF (Radial Basis Function) outperform the traditional models, MA (Moving Average), AR (Auto Regressive), and ARIMA (Auto Regressive Integrated Moving Average) in time series prediction. Neural based approaches to time series prediction require the enough length of historical measurements to generate the enough number of training patterns. The more training patterns, the better the generalization of MLP is. The researches about the schemes of generating artificial training patterns and adding to the original ones have been progressed and gave me the motivation of developing VTG schemes in 1996. Virtual term is an estimated measurement, X(t+0.5) between X(t) and X(t+1), while the given measurements in the series are called actual terms. VTG (Virtual Tern Generation) is the process of estimating of X(t+0.5), and VTG schemes are the techniques for the estimation of virtual terms. In this paper, the alternative VTG schemes to the VTG schemes proposed in 1996 will be proposed and applied to multivariate time series prediction. The VTG schemes proposed in 1996 are called deterministic VTG schemes, while the alternative ones are called stochastic VTG schemes in this paper.

• Environmental Analysis Health and Toxicology
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• 제17권3호
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• pp.239-243
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• 2002

Changes of plasma vitellogenin (VTG) and glutamate pyruvate transaminase (GPT) were examined for determining whether hepatocyte was damaged during the process of VTG induction in the juvenile rockfish, Sebastes schlegeli exposed to exogenous estrogen (estradiol-l7$\beta$, E$_2$). Rockfishes were intraperitoneally injected with E$_2$(5 mg/kg B.W.) in 70% ethanol and plasma sampling were extracted at 0, 1, 3, 6, 9, 12, 15 days af-ter E$_2$administration. VTG and GPT were then analyzed by SDS -PAGE and Reitman -Frankel method, respectively. VTG band was detected at a molecular weight position of 175 kDa on Day 3 after E$_2$administration. This band became more distinct at 6 days, but its was gradually thinned with time -course, and not detected at 15 days. GPT was suddenly increased at 1 days after 22 administration and highest GPT was detected at 3 days. However. GPT was gradually decreased with time -course as the change of VTG. These results suggest that the process of VTG induction by exogenous E$_2$damage to hepatocyte, and plasma GPT was temporarily increased in the juvenile rockfish.

• 한국양식학회지
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• 제11권2호
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• pp.241-248
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• 1998

Vitellogenin(VTG) 합성에 미치는 A23187의 영향을 무지개송어의 배양 간세포를 이용하여 실험을 행하였다. 간세포는 2일간 배양한 후, Estradiol-$17^{\beta}$($E_2$, $2{\times}10^{-6}$M) 및 A23187 ($10^{-7)$~$10^{-5}$M)을 첨가하여 7일간 배양하였다. 그리고, $E_2$에 의한 VTG 합성시의 A23187이 미치는 영향에 대해서도 조사하였다. A23187이 미치는 영향에 대해서도 조사하였다. A23187 ($10^{-7)$~$10^{-5}$M)의 첨가에 의해 간세포에서의 VTG 합성은 농도의 증가에 따라 감소하였다.$E_2$에 의해 합성된 VTG는 A 23187 ($10^{-5}$M)의 첨가에 의해 대조군($E_2$만의 첨가)의 약 18%로 유의하게 감소하였다. 그러나,$E_2$제거로는 대조군의 약 47% 밖에 감소되지 않았다. 이러한 결과로 보아, 세포내의 저장 Ca은 번역 단계 또는 번역 후 단계를 조절함으로써, VTG 합성을 조절하는 것으로 추정된다.

• Journal of Applied Biological Chemistry
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• 제52권4호
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• pp.170-173
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• 2009

Vitellogenin (Vtg) is found in the serum of both female and male fish that have been exposed to environmental endocrine disrupters or estrogen hormones, and is used as a biomarker for such contamination. In our current study antibodies raised against the purified flatfish Vtg were tested for their reactivity on immunoblots to flatfish and carp Vtg, and also to a Vtg protein fragment produced in E. coli. Polyclonal antibodies raised against purified flatfish Vtg reacted well with Vtg in the serum of flatfish and carp induced with $17{\beta}$-estradiol, but not with the Vtg protein fragment produced in E. coli.

• 한국동물학회지
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• 제38권1호
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• pp.136-144
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• 1995

참개구리의 난모세포에서 난자형성단계에 따른 vitellogenin(VTG)의 이동이 난모세포막에 의하여 조절될 것이란 생각에 기초하여 단계에 따른 VTG 이동량의 변화와 난모세포막의 변경이 VTG 이동에 미치는 영향을 조사하였다 VTG의 이동은 난자형성과정중 특히 초기난모세포에서 가장 활발하였고, 중기 및 말기에는 감소하는 경향을 나타내었다. 반면. bovine serum albumin은 모든 단계의 난오세포에서 이동을 보이지 않았다 이러한 결과로 보아 난자형성과정중 난모세포내로의 VTG 이동은 단계특이적 이고 선택적임을 알 수 있었다. 난모세포막의 막단백질을 trypsin으로 제거시키거나 막단백질의 당잔기를 wheat germ agglutinin(WGh)으로 포화시키든가 혹은 endoglycosidase $\boxDr$로 당잔기를 제거시킨 결과 VTG의 이동이 급격히 억제되었다 이와 같은 결과들은 난모세포의 막단백질이 VTG 이동에서 수용체로 작용한다는 사실을 강하게 암시하였다.