559 0

Bisphenol A 와 유사화학물질에 의한 Xenopus laevis 배아의 발생기형

Title
Bisphenol A 와 유사화학물질에 의한 Xenopus laevis 배아의 발생기형
Other Titles
Bisphenol A and their structual analogues induces developmental deformities in Xenopus laevis embryos
Author
장지현
Alternative Author(s)
Ji-Hyun Jang
Advisor(s)
계명찬
Issue Date
2021. 2
Publisher
한양대학교
Degree
Master
Abstract
The bisphenols are a group of chemicals which often used in combination with other chemicals to manufacture plastics and resins. Bisphenol A (BPA) and S (BPS) have been shown to be endocrine disruptors. Due to its high production volumes, leading to its spreading and potential accumulation in a variety of environmental matrices, even though it has a fairly short half-life. In the present study, developmental toxicity of bisphenol F (BPF) and BPS were investigated, and compared with those of BPA, using frog embryo teratogenesis assay-Xenopus (FETAX). The LC50 values of BPA, BPF and BPS were calculated to be 22.3, 50.2 and 1640.5 ìM, respectively. In surviving tadpoles, head malformation, abnormal gut coiling and ventral bilster significantly increased by BPA at 22 ìM. For BPF, head malformation significantly increased at 10 ìM. The EC50 values of BPA and BPF for malformation were 20.5 and 4.1 ìM, respectively. The teratogenic indices (LC50/EC50) of BPA and BPF were calculated to be 1.09 and 12.24, respectively. Suggest the teratogenic potential of BPF. Considering the high proportion of head malformation in surviving tadpoles, we examined the effects of bisphenols on X. laevis embryo development. X. laevis embryos at blastula stage were treated with bisphenols until neurula stage (Nieuwkoop and Faber stage 22). In bisphenols treated neurula, abnormalities such as neural tube deformity, abnormal notochord, and asymmetry were observed. The neural tube deformity was significantly increased by 22 ìM BPA. In histological analysis, the neural tube area was significantly increased by 10 mg/L BPF. The lumen area of neural tube was significantly decreased by 22 ìM BPA. The notochord area was significantly increased by 22 ìM BPA and 1 ìM BPF. The neural tube epithelia area was significantly increased by 50 ìM BPF and 1 mM BPS. We could also find an asymmetric structure at some embryo. In order to find out the reason of neural tube deformity, we determined a series of genes that regulate embryonic differentiation such as n-cadherin, e-cadherin. Cadherin, a calcium-dependent cell-cell adhesion molecule with pivotal roles in embryonic development. As the key molecule of the cadherin-catenin-cytoskeleton complex, it is important for neurulation. We tested the mRNA expression of e-cadherin and n-cadherin using RT-qPCR. As the results, the e-cadherin mRNA expression was significantly increased by 22 ìM BPA and decreased by 1 ìM BPF. In BPS treated embryo, the expression of e-cadherin mRNA was not significantly changed. The n-cadherin mRNA expression was significantly increased by 1 ìM BPA and BPS. In BPF treated embryo, the n-cadherin mRNA was significantly decreased by 50 ìM BPF. Also, since we confirmed the asymmetry of the embryos, we divided the embryos in half and sampled them separately to test the mRNA expression of xnr1, which is involved in left and right asymmetry using RT-qPCR. In bisphenols treated embryo, xnr1 mRNA expression showed abnormal expression compared to the control group. Since the previous results confirmed the abnormal expression of the xnr1 gene, the pH of the embryo was measured at st11, which determines the left and right pattern. In bisphenols treated st12 embryo had a higher pH compared to the control group using BCECF/AM. We speculated that this phenomenon was the activation of V-ATPase and measured the activity of hydrogen ions with acridine orange using microplate reader. As a result, it was confirmed that the concentration of hydrogen ions on the embryo surface was higher than that of the control group. This will lead to a series of abnormalities such as head cartilage abnormalities. As the above results can lead to deformities caused by BPF or BPS as well as BPA in the future, further research is needed. keyword : Bisphenol, Xenopus laevis, BPA, BPF, BPS, FETAX, Embryo toxicity
URI
https://repository.hanyang.ac.kr/handle/20.500.11754/159588http://hanyang.dcollection.net/common/orgView/200000485388
Appears in Collections:
GRADUATE SCHOOL[S](대학원) > LIFE SCIENCE(생명과학과) > Theses (Master)
Files in This Item:
There are no files associated with this item.
Export
RIS (EndNote)
XLS (Excel)
XML


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE