• Journal of Environmental Health Sciences
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• v.33 no.6
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• pp.513-516
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• 2007

This investigation was intended to find out the estrogenic effect of chlortetracycline (CTC) on vitellogenin induction in adult male Japanese medaka (Oryzias latipes). Vitellogenin (Vtg) produced in male fish has been used to as one of a biomarker of endocrine disrupters. The positive control was $17{\beta}-estradiol$ (E2) that induced Vtg in male fish. As a result, male and female fish were exposed to 0.1, 1, 10 and 100 ppm of CTC. Western blot results showed approximately 205 kDa, that is similar to myosin at high molecular weight range Sigma maker. Vtg band was showed fainted to 10 and 100ppm for chlortetracycline. Vtg concentration of CTC was qunatified by total protein quantification and ELISA. Exposure of the male fish to CTC of 0.1, 1, 10 and 100 ppm produced Vtg concentrations of 0.24, 0.12, 7.61 and 40.02%o, respectively, that value was elevated than control male fish (0.14%o). CTC exerted as a Vtg inducer in male fish from 10 ppm, but it was a reducer in female fish from 0.1 ppm level. The results say that vitellogenin induction patterns alter in male medaka treated with CTC, and that CTC may caused endocrine disruption in fish.

• Korean Journal of Environmental Biology
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• v.22 no.1
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• pp.206-212
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• 2004

In an effort to develop the tools for monitoring the contamination of xenoestrogen in the aquatic environment of Korea, reverse transcription-polymerase chain reaction (RT-PCR) analysis of vitellogenin (VTG) mRNA expression were optimized in Synechogobius hastus. Based on the partial VTG cDNA sequence VTG mRNA level in livers from male fishes was analyzed by RT-PCR. As an internal control beta actin mRNA was amplified. 3 ${\mu}g$ of total RNA was reverse transcribed in 20 $\mu$l reaction using murine leukemia virus 〔MuLV〕 reverse transcriptase. Subsequent PCR using the 1 ${\mu}g$ of cDNA resulted in linear increase in PCR product of VTG in female liver cDNA from 10 to 30 cycles of amplification. On the contrary, in male, PCR product first detected at 28 cycles of amplification and linearly increased during 38 cycles of amplification, suggesting that male S. hastus expresses minute amount of VTG mRNA which is $2^{-18}$ equivalent of female. In conclusion, the optimized protocol of VTG mRNA expression in the liver of male S. hastus will be promising the environmental monitoring the xenoestrogen contamination in the western coast and estuaries in Korea.

• Korean Journal of Ecology and Environment
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• v.37 no.1 s.106
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• pp.122-129
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• 2004

In an effort to develop the biomarker for monitoring the contamination of xenoestrogen in the freshwater environment of Korea, reverse transcription-polymerasechain reaction (RT-PCR) analysis of vitellogenin (VTG) gene expression was optimized in Hearisarsus Iaseo, Based on the homology of the VTG cDNA sequences between the common carp and zebra fish, a set of PCR primers for VTG mRNA amplification for H; labo was designed. VTG mRNA level in livers from female and male fishes was analyzed by RT-PCR following single injection of 17 beta estradiol($E_2$ 10 mg $kg^{-1}$ B.W.). As an internal control, beta actin mRNA was amplified. One us of total liver RNA was subjected to RT-PCR. In female the amount of PCR productof VfC gradually increased in the range from 16 to 34 cycles of amplification. On the contrary, in control male, PCR product first detected at 32 cycles of amplification and linearly increased up to 40 cycles of amplification. In $E_2$ injected male liver, the VTC mRNA level was similar to that in the female. Taken together, this result suggests that liver of male H. labo expresses minute amount of VTG mRNA which are2-l6 equivalent of female and that induction of VTG mRNA occurs in male liver after estrogen treatment. In conclusion, the optimized protocol for RT-PCR analysis of VTG mRNA expression in liver of male H. labo will provide the environmental monitoring method for the xenoestrogen contamination in the rivers in Korea.

• Environmental Analysis Health and Toxicology
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• v.28
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• pp.16.1-16.8
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• 2013

Objectives Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system. Methods Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations. Results The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was $99.5{\pm}5.5%$. Conclusions A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.

• Journal of Aquaculture
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• v.15 no.1
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• pp.31-37
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• 2002

Effect of 2,4-Dichlorophenoxy acetic acid (2,4-D) on vitellogenin (VTG) production and estrogen ($E_2$)-estrogen receptor (ER) binding affinity were examined in primary hepatocyte cultures of rainbow trout, Oncorhynchus mykiss. Hepatocytes were pre-cultured for 2 days; subsequently, $E_2$( 2$\times$$10^{-6}$/ M) and 2,4-D ($10^{-9}~10^{-6}/M$) were simultaneously added to the incubation medium. They were cultured for more than 5 days. VTG and $E_2$-ER binding affinities were analyzed by SDS-PAGE and ELISA, respectively. 2,4-D concentration used had no appreciable effect on the morphology, viability, and DNA content of hepatocytes in culture. It had also no effect on VTG production. However, it interfered with $E_2$-ER binding affinity, which was reduced with increasing concentration of 2,4-D. The affinity was inhibited by 25 and 30% at $10^{-7}$ M and $10^{-6}$ M of 2,4-D, respectively. This result suggested that although 2,4-D had no effect on VTG production, it acted as reno-estrogenic contaminant in ER.

• Fisheries and Aquatic Sciences
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• v.5 no.4
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• pp.251-257
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• 2002

Effects of bisphenol-A (BPA) on vitellogenin (VTG) synthesis and estrogen-estrogen receptor $(E_2- ER)$ binding activity were examined in primary hepatocyte cultures of rainbow trout, Oncorhynchus mykiss. Hepatocytes were precultured for 2 days and then $estradiol-17\beta\;(E_2,\;2\times10^{-6}M)\;BPA\;(10^{-5}-10^{-8}M)$ and/or 4-hydroxy-tamoxifen $(4-OHT,\;10^{-6} M)$ were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days and then spent medium was analyzed by SDS-PAGE for VTG production. The addition of BPA to the incubation medium had no effect on the viability of hepatocytes in the culture. On the other hand, BPA increased VTG production in a concentration-dependent way and a significant increment occurred at BPA concentrations greater than $10^{-6}$M. Although VTG was increased by the addition of $E_2\;(2\times10^{-6}\;M)\;or\;BPA\;(10^{-5}M)$, its were reduced by a simultaneous 4-OHT $(10^{-6}\;M)$ addition. BPA inhibited $E_2-human$ ER binding activity by $72\%$ at $10^{-5}$ M of BPA. These results suggested that BPA induced VTG synthesis by BPA-ER binding activity in the hepatocyte of rainbow trout.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2003.05a
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• pp.156-157
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• 2003

In teleost, vitellogenin (Vtg) is synthesised in the liver in response to estrogens and transported by the blood to the growing oocytes where it is incorporated by micropincytosis (Selman and Wallace, 1982). Generally, Vtg is not induced in normal male fish but male fish are capable of synthesis Vtg in reponse to exogenous estrogen and xenoestrogens. (omitted)

• Journal of Aquaculture
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• v.11 no.2
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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.

• Journal of Korea Multimedia Society
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• v.22 no.3
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• pp.357-365
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• 2019

In this paper, we suggest the tracking method of moving multi-objects in maritime environments. The image acquisition is conducted using IR(InfraRed) camera sensors on an airborne platform. Under the circumstance of maritime, the qualities of IR images can be significantly degraded due to the clutter influence, which directly gives rise to a tracking loss problem. In order to reduce the effects from the clutters, we introduce a technical approach under Man-In-The-Loop(MITL) system for enhancing the tracking performance. To demonstrate the robustness of the proposed approach based on VTG(Valid Tracking Gate), the simulations are conducted utilizing the airborne IR video sequences: Then, the tracking performances are compared with the existing Kalman Filter tracking techniques.

• Environmental Analysis Health and Toxicology
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• v.22 no.4
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• pp.321-327
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• 2007

4-nonylphenol (4-NP)이 해산어류인 조피볼락, Sebastes schlegeli의 혈장 vitellogenin (VTG), alkaline-labile protein phosphorus (ALPP), calcium(Ca), glutamate pyruvate transaminase (GPT) 및 hepatosomatic index (HSI)에 미치는 영향을 조사하였다. 실험어에 3일간격으로 $estradiol-17{\beta}$ ($E_2$, 5 mg/kg B.W.) 또는 4-NP(0, 10, 50, 100및 200 mg/kg B.W.)을 복강에 2번 주사한 후, 7일째에 채혈과 적출을 통해 혈장과 간장을 수집해 분석이 실시되었다. 대조 실험어에는 용매로 사용된 70% 에탄올만이 투여되었다. $E_2$ 투여 실험어의 혈장 단백질을 전기 영동상으로 분석한 결과 약 170 kDa의 위치에서 짙은 VTG 밴드가 관찰되었으나, 용매만 투여한 대조 실험어의 혈장에서는 동일 밴드가 관찰되지 않았다. 4-NP 투여한 모든 실험어의 혈장 단백질에서는 $E_2$ 투여 실험어와 동일한 VTG 밴드가 관찰되었다. 혈장 ALPP와 Ca 농도도 4-NP 투여 실험어에서 $E_2$ 투여 실험어와 유사하게 증가하였으며, 이들 농도 변화는 VTG 합성과 더불어 증가하는 경향을 나타냈다. 혈장 전위효소인 GPT와 HSI도 $E_2$ 투여 실험어와 유사하게 4-NP가 투여된 모든 실험어에서 급격히 증가하였다. 이상의 결과로부터 연안생태계 내에서 서식하는 어류가 4-NP과 같은 내분비 장애물질(Endocrine Disrupting compounds, EDCs)에 의해 영향을 받는지를 규명하기 위한 생물학적 지표로서 VTG와 더불어 혈장 ALPP와 Ca이 사용가능 할 것으로 판단된다. 또한, 조피볼락과 같은 해산어가 EDCs에 노출되어 VTG가 합성될 때 간장 기능의 손상으로 혈장 전위효소인 GPT가 일시적으로 종가하고 간장도 비대해져 HSI가 높아지는 것으로 판단된다.