华东师范大学学报(自然科学版) ›› 2007, Vol. 2007 ›› Issue (4): 95-100.

• 生命科学 • 上一篇    下一篇

HgCl2与CdCl2对河蟹精子DNA损伤的单细胞凝胶电泳检测研究

范丽君, 周忠良, 陈东华, 赵云龙, 孙菊香, 王 群   

  1. 华东师范大学 生命科学学院, 上海 200062
  • 收稿日期:2006-03-03 修回日期:2006-03-30 出版日期:2007-07-25 发布日期:2007-07-25
  • 通讯作者: 王 群

Study on DNA Damage Induced by HgCl2 and CdCl2 Using SCGE in Eriocheir Sinensis Sperms (In Vitro)(Chinese)

FAN Li-jun, ZHOU Zhong-liang, CHEN Dong-hua , ZHAO Yun-long, SUN Ju-xiang, WANG Qun
  

  1. School of Life Science, East China Normal University, Shanghai 200062,China
  • Received:2006-03-03 Revised:2006-03-30 Online:2007-07-25 Published:2007-07-25
  • Contact: WANG Qun

摘要: 采用单细胞凝胶电泳技术,研究了HgCl2和CdCl2对河蟹(Eriocheir sinensis)精子DNA损伤的影响.HgCl2和CdCl2各设置6个相同的浓度梯度,分别为0 mmol/L,0.01 mmol/L,0.05 mmol/L,0.1 mmol/L,1 mmol/L,5 mmol/L,处理温度为37 ℃(水浴),处理时间为1 h;SCGE的检测指标为DNA拖尾长度和DNA损伤率.结果表明,对照组DNA未受损伤,在电场中核DNA几乎不泳动,染色后呈圆形的荧光团,无拖尾现象;各处理组精子DNA均不同程度受损,电泳染色后均呈现彗星状荧光团,且随着HgCl2和CdCl2浓度的升高,河蟹精子DNA迁移距离和损伤率亦随之增加,DNA损伤程度与其浓度之间存在明显的“剂量-效应”关系.此外,本实验亦证实SCGE可用于河蟹精子DNA损伤的检测.

关键词: 中华绒螯蟹, 河蟹, 单细胞凝胶电泳, 氯化汞, 氯化镉, 精子DNA, 中华绒螯蟹, 河蟹, 单细胞凝胶电泳, 氯化汞, 氯化镉, 精子DNA

Abstract: DNA damage induced by mercuric chloride (HgCl2) and cadmium chloride (CdCl2) in Eriocheir sinensis sperms (in vitro) was studied using single-cell gel electrophoresis (SCGE). Separated DNAs were dealt with six HgCl2 and CdCl2 scalar solutions (for each) (0 mmol/L, 0.01 mmol/L, 0.05 mmol/L, 0.1 mmol/L, 1 mmol/L, 5 mmol/L) for one hour at 37 ℃ (water bath) respectively. The length of cometic tail and damage rate of DNAs were betaken as SCGE indices. Results showed that DNAs in control group are free of damage, static in electric field and manifested as a round fluorophoreu without any cometic tails. While those dealt DNAs are damaged to various degrees and demonstrated as cometic fluorophoreu in electric field after Ethidium bromide (EB) stain. Furthermore, with the incremental concentration of HgCl2 and CdCl2, damage rate and migration distance also enlarge accordingly, that′s to say, dose-response relationship exist between SCGE indices and concentration of HgCl2 and CdCl2. Therefore, it is concluded that certain concentration of HgCl2 and CdCl2 does induce DNA damage in Eriocheir sinensis sperms (in vitro) and the damage degree is related to the concentration of HgCl2 and CdCl2, which have been attested using SCGE.

Key words: sperms DNA, SCGE, HgCl2, CdCl2, Eriocheir sinensis, sperms DNA, SCGE, HgCl2, CdCl2

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