Study on DNA Damage Induced by HgCl2 and CdCl2 Using SCGE in Eriocheir Sinensis Sperms (In Vitro)(Chinese)
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摘要: 采用单细胞凝胶电泳技术,研究了HgCl2和CdCl2对河蟹(Eriocheir sinensis)精子DNA损伤的影响.HgCl2和CdCl2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HgCl2和CdCl2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.Abstract: DNA damage induced by mercuric chloride (HgCl2) and cadmium chloride (CdCl2) in Eriocheir sinensis sperms (in vitro) was studied using single-cell gel electrophoresis (SCGE). Separated DNAs were dealt with six HgCl2 and CdCl2 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 HgCl2 and CdCl2, damage rate and migration distance also enlarge accordingly, that′s to say, dose-response relationship exist between SCGE indices and concentration of HgCl2 and CdCl2. Therefore, it is concluded that certain concentration of HgCl2 and CdCl2 does induce DNA damage in Eriocheir sinensis sperms (in vitro) and the damage degree is related to the concentration of HgCl2 and CdCl2, which have been attested using SCGE.
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Key words:
- Eriocheir sinensissperms DNASCGEHgCl2CdCl2 /
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