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Xenopus laevis 초기 배의 동물극 분리배양에서 bFGF와 HGF 혼합처리에 의한 기관유도
진정효,윤춘식,이호선,박용욱,정선우 한국생명과학회 2004 생명과학회지 Vol.14 No.3
FGFs는 Xenopus의 초기 배발생에서 중배엽 형성, 전후축패턴형성, scatter factor로서 낭배기의 기관형성에 관여하는 등 다양한 기능을 가지고 있는 것으로 알려져 있다 그 중 bFGF는 배양 분리편으로부터 다양한 기관을 유도해낼 수 있으며 그 효과는 처리시간 및 농도 의존적이라고 알려져 왔다. 본 연구는 Xenopus의 예정표피역을 분리하여 bFGF와 HGF을 단독 및 복합처리 하였을 때 기관분화 및 유도효과를 검토하기 위하여 실시하였다. 단독처리 및 복합 처리된 배양액에 동물극 분리편을 정상배가 st. 43에 이를 때까지 2$0^{\circ}C$에서 3일간 배양하여 조직학적 및 면역조직화학적 방법으로 조직의 분화양상을 확인하였다. 성장인자는 bFGF를 0, 0.5, 1. 10. 50 ng/ml의 농도와 HGF를 0, 1, 10, 50, 100ng/ml의 농도로 조합하여 처리한 결과 bFGF 단독처리 때보다 HGF와의 혼합처리에서 기관분화율의 상승효과가 관찰되었다. 분화된 기관은 1 ng/ml의 bFGF 와 10ng/ml의 HGF, 10ng/ml의 bFGF와 1ng/ml의 HGF의 농도에서 매우 다양한 것으로 나타났다. 눈은 1과 10ng/ml의 bFGF ,그리고 1과 10 ng/ml의 HGF 농도조합에서 높은 비율로 분화하였다. 또한 분리편 배양에 의해 유도된 눈과 정상 배의 눈에서 RPE65를 인식하는 단일클론 항체 40All, 25F5를 사용하여 AP 반응이 강하게 나타나 눈의 유도를 확인할 수 있었다. Fibroblast growth factors (FGFs) are known to induce multiple functions in early development, including mesoderm formation, gastrulation movement and antero-posterior patterning. The induction of mesoderm from Xenopus presumptive ectoderm and the combination effect on inducing organs of bFGF(basic FGF) and HGF (Hepatocyte Growth Factor) were studied. Explants were cultured in the combined solution for 3 days to normal embryo arrive at St. 43. These effects on combined dose were examined by histological experiment and by immunohistochemical method. The concentrations of growth factors were tested in 0, 0.5, 1, 10 and also tested in 50 ng/ml of bFGF, and 0, 1, 10, 50 and 100ng/ml of HGF respectively. The synergistic effects were seen in the combined-dose of bFGF and HGF rather than in single dose. Various organs were differentiated and highest inducing effects were seen at the combined concentration of 1 ng/ml of bFGF and 10ng/ml of HGF, and at the concentration 10ng/ml of bFGF and 1 ng/ml of HGF. The bFGF induces various organs from cultured animal cap explants and the effects are time and dose-dependent. HGF is also a potent mitogen for renal tubular cells and for mature hepatocytes in primary culture. Eyes were developed in high percentage at the combined concentration of 1 and 10ng/ml of bFGF, and 1 and 10 ng/ml of HGF. From the induced eye and normal embryonic eye, RPE65 was commonly detected by monoclonal antibodies 40All and 25F5 and the localization of RPE65 was seen by AP reaction.
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정선우,진정효,전주아 昌原大學校 基礎科學硏究所 2000 基礎科學硏究所論文集 Vol.12 No.-
The insect fauna from the district of Mt. Hwaak was studied. Specimens were collected from April to October of 1998. The total specimens were classified into 9 orders, 70 families, and 300 species. In Dermaptera, 1 families 1 species were identified. In Odonata, 1 families 3 species were collected, and 7 families 13 species were contained in Homoptera. In Hemiptera, 11 families 22 species were classified, and 5 families 10 species were classified in Orthoptera. Diptera held 6 families 9 species, Hymenoptera had 8 families 20 species, and Coleoptera had 8 families 44 species. And we could classified 23 families 176 species in Lepidoptera.
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과량의 아연에 의한 아프리카 발톱개구리 (Xenopus Laevis)의 배발성 이상과 독성
윤춘식 ( Chun Sik Yoon ),진정효 ( Jung Hyo Jin ),정선우 ( Seon Woo Cheong ) 한국하천호수학회 2003 생태와 환경 Vol.36 No.1
Concentrated releases of zinc into water usually results from discharges associated with industrial purpose. The released zinc into soil is corroded and released into water. In aquatic environment, exess zinc is toxic to the organisms and causes the growth inhibition and malformation of them as a heavy metal. In this study, excess zinc toxicity was tested by FETAX (frog embryo teratogenetic assay with Xenopus) as in vivo system. Xenopus embryos at st. 9 were exposed to 100~900μM of zince for 7 days and 81% of individuals were survived in 100μM, and 25% were survived in 1000 M of zinc solution. In external malformations, swelled belly and intestinal dysplasia were common, and all of tested individuals showed these malformations in 200μM or higher concentration of zinc. In 400μM or higher concentration, all of tested tadpoles showed faded heart. Also, hypo-pigmentation, lens hernia and loose digestive track were very frequently found in 100μM of zinc. The histological study with paraffin section of zinc treated tadpoles showed following abnormalities; regeneration of photoreceptor on retina, reduced vitreous chamber in eye, reduction of red blood cells in heart, abnormal liver, swelling of pronephric cell, muscle dysplasia and palatal papilloma. These abnormalities may be caused by the degeneration of mitochondria, inhibition of cell adhesion, and the formation of leghemoglobin by zinc due to the substitution of Ca^2+ by Zn^2+. The body length was reduced due to the excess zinc. From a statistical result, body lengths of 300μM or higher concentrative groups was significantly reduced comparing that of control group. Recently, many spontaneous malformations and reduction of amphibians are reported. Form the results of present study, excess zinc might be a factor of amphibian reduction, and the control of zinc discharges is very important.
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한국산 도롱뇽 ( Hynobius leechii ) 의 농경지에서의 배 발생 이상과 살균제 Benomyl 의 독성효과
최영주(Yeoung Ju Choi),윤춘식(Chun Sik Yoon),박주홍(Joo Hung Park),진정효(Jung Hyo Jin),정선우(Seon Woo Cheong) 한국하천호수학회 2002 생태와 환경 Vol.35 No.3
A numerical variation and abnormalities were studied on egg bags and embryos of Korean salamander, Hynobius leechii from agricultural habitat. The teratogenic and toxic effects of fungicide benomyl were also investigated with early embryos from non-agricultural habitat. We collected 144 egg bags from agricultural region, and 3418 of early embryos were contained. The lengths of egg bags were varied from 10 to 23㎝ and the most frequent length was 19㎝. The number of embryos was varied from7 to 43, and the most frequent range was 22 to 26. Spontaneous abnormalities were occurred in 406 embryos among 116 egg bags, and 24 kinds of external abnormalities were found. Individuals showing severe external defect were histologically studied and they showed optic dyspalsia, thyroid carcinoma, somatic muscular dysplasia, partial biaxial structure, decrease of red blood cells in the heart, cephalic degeneration and intestinal dysplasia. 385 embryos from non-agricultural region were exposed to 200nM∼1μM of benomyl at blastula or gastrula for 12 days. All embryo were dead in the concentration of 1μM(LD_100) and 75% of embryos were dead in 800 nM of benomyl. Specific effect due to benomyl was acrania or cephalic dysplasia and this result suggests that the benomyl inhibit strongly to the development of neural tissue. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of the movement of neural crest cells.
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