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目的探讨铅对斑马鱼胚胎发育的毒性作用。方法野生型AB品系斑马鱼胚胎,自受精后1h(1hpf)起,分别用醋酸铅质量浓度为0.1、0.5、2.5、12.5μmol/L胚胎培养液染毒至胚胎孵出或死亡,每组胚胎50枚,相差显微镜下观察胚胎发育全程并记录毒性反应,于4、8、16、24、48、72hpf进行胚胎活体摄片,以不含铅培养液孵养的胚胎作为对照。另取各组36 hpf胚胎,电镜下观察间脑神经细胞超微结构。结果铅暴露组胚胎死亡率明显升高(P<0.05),在胚胎发育早期有1个高峰,而且随着铅暴露浓度增高,胚胎死亡率总体呈上升趋势(P<0.01);铅暴露胚胎发育异常主要见于胚胎发育早期,多表现为胚胎发育迟缓;铅暴露胚胎在发育过程中出现少数畸形,但无统计学意义;除12.5μmol/L铅暴露组外,其他铅暴露组胚胎的平均孵出时间较对照组滞后(P<0.05);电镜观察显示,铅暴露可引起胚胎神经细胞核固缩,细胞水肿,细胞器结构受损。结论铅对斑马鱼胚胎发育有明显的毒性作用,在胚胎发育早期尤为敏感;即使较低水平的铅暴露,仍可引起胚胎神经细胞的损害。
Objective To explore the toxic effect of lead on zebrafish embryo development. Methods The wild-type AB strain zebrafish embryos were incubated at 1h (1hpf) after fertilization and incubated with 0.1, 0.5, 2.5, 12.5μmol / L lead acetate solution for embryo incubation. The whole embryos were observed under phase contrast microscope and the toxicity was recorded. Live embryos were taken at 4, 8, 16, 24, 48 and 72 hpf. Embryos incubated with Pb-free medium were used as controls. Another group of 36 hpf embryos were observed under electron microscopy ultrastructure of cerebral neurons. Results The embryo mortality rate was significantly higher in lead exposed group (P <0.05), with a peak in early embryo development, and the embryo mortality rate increased with the increase of lead exposure concentration (P <0.01) Abnormalities were mainly found in the early embryo development, mostly in embryonic development retardation; lead exposed embryos in the development process of a small number of abnormalities, but no statistical significance; except 12.5μmol / L lead exposure groups, other lead exposure group embryos average hatched Compared with the control group, the time lagged (P <0.05). Electron microscopy showed that lead exposure induced nuclear pyknosis, cell edema and organelle structure damage in embryonic nerve cells. Conclusion Lead has significant toxic effects on the development of zebrafish embryos and is particularly sensitive to early embryo development. Even low levels of lead exposure may cause damage to embryonic nerve cells.