EE2对稀有鮈鲫和斑马鱼幼鱼体内卵黄蛋白原诱导的比较
COMPARATIVE WTELLOGENIC RESPONSES IN ZEBRAFISH(BRACHYDANIO RERIO) AND RARE MINNOW(GOBIOCYPRIS RARUS) EXPOSED TO 17α-ETHINYLESTRADIOL
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摘要: 利用卵黄蛋白原(Vtg)作为类雌激素污染的生物标志物,比较研究了不同浓度的17α-乙炔基雌二醇(EE2)对斑 马鱼(Brachydanio rerio)和稀有鮈鲫(Gobiocypris rarus)幼鱼体内Vtg的诱导。研究结果表明:5ng/L,20ng/L和100ng/L EE2分别暴露5d后,稀有鮈鲫幼鱼体内的Vtg即可显著诱导,并且其含量随暴露时间的增加而增加,在暴露15d时 达到最大值;而斑马鱼幼鱼虽然100ng/LEE2暴露5d时可显著诱导体内Vtg的生成,但20ng/L EE2在暴露10d后, 5ng/L EE2在暴露15d后,才可显著诱导斑马鱼体内Vtg的生成。这一结果说明EE2暴露下对稀有鮈鲫体内Vtg的 诱导要比斑马鱼敏感。Abstract: In this study, the response of zebrafish and rare minnow to 17α-ethinylestradiol(EE2) was studied using vitellogenin (Vtg)as a biomarker. Juvenile rare minnow and zebrafish were exposed to waterborne EE2 at 5ng/L,20ng/L and 100ng/L EE2 in laboratory aquaria. After the exposure of 5, 10, 15 days, the fish were sampled to measure Vtg in whole body homogenates (WBH) .The results showed that Vtg levels in juvenile rare minnow increased significantly at 5ng/L,20ng/L and 100ng/L EE2 after 5 days exposure and reached a maximal level on the day 15th. Although Vtg concentration in juvenile zebrafish increased significantly in 100 ng/L EE2 exposure group after 5 days,a significant increase of Vtg in 5ng/L and 20ng/L treatment was just found on the day 15th and on the day 10th respectively. These indicate that rare minnow is more sensitive than zebrafish in Vtg induction to 17α-ethinylestradiol. Zebrafish Vtg was identified in this study. After being separated by SDS-PAGE, zebrafish Vtg produced two protein bands with molecular weights of 140 kDa and 170kDa. Because the development of species-specific methods for its quantification has been hampered due to problems such as the small blood volume and the lack of sufficient pure antigen and specific antibodies, a polyclonal antibody raised against carp Vtg was used as first antibody to measure Vtg in whole body homogenates of zebrafish and rare minnow in this study.The results of western blotting showed that the carp Vtg antibodies cross-reacted with zebrafish Vtg and could be used to measure zebrafish Vtg.
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Keywords:
- Rare minnow /
- Zebrafish /
- 17α-ethinylestradiol /
- Vitellogenin
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