亚甲基蓝对黄颡鱼鱼卵高卵壳裂解酶(HCE)表达的影响

鱼卵高卵壳裂解酶(HCE)是鱼类孵化酶中的一种,在水生生物胚胎发育过程中起非常重要的作用,其表达可能会受到化学药物的影响。以黄颡鱼(Pelteobagrus fulvidraco)受精卵为研究对象,采用免疫组织化学方法研究了亚甲基蓝对黄颡鱼鱼卵高卵壳裂解酶表达的影响。以15 mg/L亚甲基蓝浸泡黄颡鱼受精卵,同时设置不做任何药物处理的空白对照组,分别在受精后0、1、2、4、8、12、24 h取黄颡鱼受精卵或胚胎,4%多聚甲醛固定,石蜡切片,免疫组织化学方法染色,倒置显微镜观察,Image-Pro Plus6.0软件分析每张照片的累积光密度值。结果表明,亚甲基蓝实验组与空白对照组中均有HCE表达,但是亚甲基蓝实验组中HCE的相对表达量显著高于其相应的空白对照组。不同受精时间的对照组和实验组之间HCE的相对表达量无统计学差异。亚甲基蓝明显促进HCE的分泌,可能与鱼卵的代偿或应激反应有关,由此推测亚甲基蓝对胚胎有一定的毒性。

The effect of methylene blue on the expressions of high choriolytic enzyme in the embroyo of Pelteobagrus fulvidraco

High choriolytic enzyme (HCE) plays an important role during the fish artificial propagation,which could be affected by chemical drugs.In this study,the embryo of Pelteobagrusfulvidraco was chosen to measure the effect of methylene blue on expressions of HCE by using immunohistochemical method.The zygotes of P.fulvidraco was steeped with 15 mg/L methylene blue,meanwhile the blank control group was steeped with no drug conduct.The zygotes or embryos of P.fulvidraco which have fertilized 0 h,1 h,2 h,4 h,8 h,12 h,24 h were selected,immobilized by 4％ paraformaldehyde and make paraffin section.Each section was observed under the microscope after immunohistochemical staining.The integral optical density(IOD) of each photograph was analyzed by Image-Pro Plus6.0 software.The results revealed that hatching enzyme distributed both in methylene blue experimental group and blank control group.However,the relative expression of HCE in methylene blue experimental group was significantly higher than the blank control group.There was no statistics difference between the relative expression of HCE in blank control group and experimental group.The results of showed that methylene blue could significantly promote the secretion of HCE.It may be associated with the compensatory or stress response of fish embryos.So we can infer that methylene blue has toxicity to embryos.