• 환경생물
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• 제22권2호
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• pp.300-307
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• 2004

The chloroacetanilide herbicide alachlor (2-chloro-2', 6'-diethyl-N-(methoxymethyl)-acetanilide) and organochlorine insecticide endosulfan (6, 7, 8, 9, 10, 10-hexachloro-l, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 3, 4-benzodioxathiepin-3-oxide)are the highly toxic agricultural chemicals. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. B. orientalis spawns in the farming regions at Spring when the massive application of agricultural chemicals occurs. These chemicals in farmland may threaten the reproduction of this frog. Therefore, we examined the embryotoxic and survival effects of alachlor and endosulfan at various concentrations in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with alachlor and endosulfan were decreased with concentration dependent manner. Also, developmental malformations appeared by alachlor and endosulfan in B. orientalis embryos. The malformations showed in order of frequency with bent trunk, tail dysplasia, bent tail, thick-set body and ventral blister in alachlor treated embryos. The exposure of endosulfan produced 7 types of severe external malformations with tail dysplasia, pectoral blister, bent trunk, bent tail, cephalic dysplasia, ventral blister, and thick-set body. Following exposure to alachlor and endosulfan the types of malformations were diverse, suggesting these chemicals target multiple events in embryonic and larval development in this species. These results suggested that alachlor and endosulfan were detrimental for survival and development of B. orientalis embryos.

• 환경생물
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• 제29권1호
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• pp.74-80
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• 2011

In this study we present a mammalian cell culture model that allows to study the effect of endocrine disruptors (EDCs) on aromatase activity of aquatic amphibian, Bombina orientalis. Bombina orientalis aromatase gene was subcloned into a mammalian expression vector and subsequently transfected to mammalian cells. Although the protein expression level of Bombina orientalis aromatase was low, it had a significant aromatase activity. When EDCs were added to aromatase transfected cells, aromatase activity was significantly decreased. We report here that this system may be used to monitor the effect of EDCs on aromatase activity of aquatic organisms.

• 대한화학회지
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• 제18권5호
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• pp.368-372
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• 1974

Bombina Orientalis(무당개구리)의 복피로부터 추출한 암적색의 색소를 TLC, PLC(preparative thin layer chromatography) 및 column chromatography로 분리 및 정제하여 물리, 화학적 성질 visible 및 IR-spectral 특성을 이용하여 세번째주색소인 ${\alpha}$-cryptoxanthin(3-hydroxy-${\alpha}$-carotene)을 확인하였다

• 한국환경생물학회:학술대회논문집
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• 한국환경생물학회 2004년도 학술대회
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• pp.19-19
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• 2004

• 대한화학회지
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• 제18권4호
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• pp.289-296
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• 1974

Bombina Orientalis(무당개구리)의 복피로부터 암적색의 색소를 추출하여 TLC 및 column chromatography 로 10여종의 carotenoid를 분리하여 visible 및 IR-spectrum을 조사하였다. 물리, 화학적 성질, chromatomatographic 성질 visible 및 IR-spectral 특성을 이용하여 주색소는 ${\beta}$-carotene과 4-hydroxy-echinenone임을 확인하였다.

• 환경생물
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• 제26권1호
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• pp.22-29
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• 2008

무당개구리(Bombina orientalis, Korean fire-bellied toad)는 양서강 개구리목 무당개구리과 무당개구리속(Class Amphinia, Order Anura, Family Bombinatoridae Genus Bombina)의 무미양서류이다. 소형으로 행동이 느리고 온순하여 취급이 용이하며 실험모델로써 많은 장점이 있으나 번식기를 제외한 개체를 대상으로 한 연구는 시료확보의 난점으로 무당개구리의 계절번식 특성에 대한 연구는 미진하다. 최근 지구온난화의 직간접적인 영향으로 전 세계적으로 무당개구리를 포함한 많은 양서류가 감소하고 있으며 야생채집개체를 활용하는 연구는 점차 규제를 받을 것으로 사료된다. 이전 연구에서 시행되었던 바이오톱 조성 목적과 달리 양서류 번식생물학적 연구를 위한 자연서식환경 조성은 시도되지 않았다. 본 연구는 무당개구리의 생식, 발생, 생태연구를 실험실 환경에서 수행하기 위한 목적으로 자연서식환경을 모방한 biotope 계획과 조성지침, 그리고 표준처리절차(SOP)를 제시하였다. 조성 후 시험적으로 방사한 개체연구결과로 는 실내사육실에서 키웠던 개체와는 달리 자연생태환경에서 관찰되는 계절적 적응현상을 관찰할 수 있었다. 본 지침서를 기초로 조성된 양서류 biotope을 활용하면 온대지방에 서식하는 양서류의 계절번식 생태를 실험실 환경에서 연구하는 데 유용할 것으로 사료된다.

• Applied Microscopy
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• 제28권3호
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• pp.329-343
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• 1998

Early wound healing responses in the skin of the Korean fire bellied toad, Bombina orientalis, were examined using histochemical , scanning and transmission electron microscopical techniques. Cutaneous wounds were induced by sharp razor blade , and maintained in special cages for up to 3 days. By this treatment whole parts of epithelium and dermis were profoundly affected. After injury, retraction of damaged surface of the skin was observed. At 3 hours after injury, poly-band was form by mucous substance. The wound skin consisted of mucous gland secreted mucous substance which flows into wound cavity and the wound area was sealed by mucous substance which form thick layer at 6 hours. It appeared that natural disposition showed acid and neutral mucous substance by reaction of PAS and Alcian blue (pH 2.5) staining. After formation of the poly-band, epithelial cells were formed by migration of regenerating epithelial cells. These cells moved over the wound surface by migration at 12 hours after wounding. Poly-band was much increased in thickness and migrating cells were completely formed wound closure at 24 hours post-wounding. After 72 hours regenerated epithelial layer consists of four or five cell layers which are similar to clear, granular, and spinous layer.

• Applied Microscopy
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• 제28권3호
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• pp.379-390
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• 1998

Dermal wound healing responses in the skin of the toad, Bombina orientalis, were examined using transmission electron microscopy. At 12 hours after wounding, debridement and collagenolysis occurred in damaged dermis. Histocyte has a large nucleus and long cytoplasm process. Phagocytic vesicle and lysosome were observed in the cytoplasm. Damaged blood cells were transformed spindle to irregular shape. Autolysis was observed in their cytoplasm. Histocytes are found in poly-band. The irregularly shaped nucleus is located peripheral region in cytoplasm. At 2 days after wounding, partial aggregation of blood cells is observed. Phagocytic, activity is observed in histocyte and collagenolytic collagen fibers are scattered. Fibroblast is observed in the dermis at 3 days after wounding. Clusters of ribosomes and some short cisternae of rough-surfaced endoplasmic reticulum are found in the cytoplasm. In histocyte at 7 days post wounding, various size granules composed of moderately dense material are found the cytoplasm. In this period histocyte is round to rod in profile, with slender processes projecting from the surface. At 7 day after injury, it was observed that formation of connective tissue fibers and amorphous ground substance in regenerating skin.

• 한국동물학회지
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• 제29권4호
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• pp.261-271
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• 1986

무당개구리(Bombina orientalis Boulenger)背皮의 청록색 발현에 관여하는 黃色素細胞(xanthophore)의 분화과정을 광학 및 전자 현미경으로 관찰하였다. 성체의 개구리에서 관찰되는 황색소세포는 pterinosome과 carotenoid vesicle등 2가지 색소과립을 함유하고 있으며, 진피의 다른 색소세포들과 複合體를 이루어 기저막을 따라 길게 세포질의 돌기를 뻗고 있었다. 최초의 황생소세포는 幼生期의 섭식시기를 전후하여 진피층에 나타나는 rER이 풍부한 세포와 Golgi complex가 풍부한 세포로부터 發生, 分化하며 변태의 각 단계를 거치면서 점진적으로 색소과립의 형성이 이루어진다. 변태가 완료되는 시기를 전후하여 pterinosnme의 급격한 증가현상이, 그리고 동면기 이후에는 carotenoid vesicle의 증가 현상이 관찰되는데, 이들 색소과립은 분화의 정도에 따라 여러 가지 구조를 나타내었다.

• 환경생물
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• 제22권2호
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• pp.329-335
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• 2004

To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription -polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using $1\;\mu{g}$ of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT- PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a $2^{-12}$ equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.