›› 2014, Vol. 57 ›› Issue (2): 194-203.

• 研究论文 • 上一篇    下一篇


吴艳艳, 周婷*, Abebe Jenberie WUBIE, 王强, 代平礼, 贾慧茹   

  1. (中国农业科学院蜜蜂研究所, 农业部授粉昆虫生物学重点实验室, 北京 100093)
  • 出版日期:2014-02-20 发布日期:2014-02-20

Apoptosis in the nerve cells of adult honeybee (Apis mellifera ligustica) brain induced by imidacloprid

WU Yan-Yan, ZHOU Ting*, Abebe Jenberie WUBIE, WANG Qiang, DAI Ping-Li, JIA Hui-Ru   

  1. (Key Laboratory of Pollinating Insect Biology, Ministry of Agriculture, Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China)  
  • Online:2014-02-20 Published:2014-02-20

摘要: 【目的】蜜蜂是重要的授粉昆虫, 但一直受到新烟碱类杀虫剂(如吡虫啉)的危害, 该类杀虫剂主要作用于蜜蜂脑神经细胞。本研究旨在明确吡虫啉对意大利蜜蜂Apis mellifera ligustica脑神经细胞致凋亡作用。【方法】利用亚致死剂量吡虫啉饲喂成年意大利蜜蜂, 通过原位末端标记法(TUNEL)检测脑神经细胞的凋亡情况, 利用免疫荧光法检测Caspase-1激活情况, 通过透射电镜观察脑神经细胞的超微结构。【结果】在对选取的脑部7个部位进行总体观测后, 摄入亚致死剂量的吡虫啉(9.90 ng/蜜蜂)诱导成年意蜂工蜂脑神经细胞凋亡率随时间递增而增加, 在处理9 d和12 d时, 处理组与空白对照组差异极显著(P<0.01); Caspase-1阳性细胞率也随时间递增而增加, 在处理3, 6, 9和12 d时, 处理组与空白对照组差异极显著(P<0.01)。凋亡率和Caspase-1阳性细胞率均表现出时间效应, 且两者成正相关性。超微结构表明凋亡的神经细胞呈现凋亡和自噬双重特征, 包括: 细胞固缩、 染色质浓缩、 凋亡小体和自噬体出现; 线粒体肿胀, 有的被自噬泡包裹后进行线粒体自噬。【结论】本研究从细胞水平证实了亚致死剂量吡虫啉对成年意蜂工蜂脑神经细胞具有致凋亡作用, 并且其凋亡途径与Caspase-1和细胞自噬有关, 这为利用细胞凋亡法评价杀虫剂对非靶标生物的神经毒性提供一定的理论基础。

关键词: 意大利蜜蜂, 新烟碱类杀虫剂, 吡虫啉, 细胞凋亡, 自噬, Caspase依赖型, 超微结构

Abstract: 【Aim】 Honeybees are important pollinators, but they are endangered by neonicotinoid insecticides like imidacloprid which mainly impair nerve cells of honeybee brain. Our research aimed to explore the apoptosis in the nerve cells of Apis mellifera ligustica brain induced by imidacloprid. 【Methods】 Apoptosis was detected in the nerve cells of honeybee brain by TUNEL labeling, cleaved Caspase-1 was detected by immunofluorescence, and the ultrastructure of nerve cells in the brain of adult worker honeybees was observed by transmission electron microscope after treatment with the sublethal dose of imidacloprid (9.90 ng/individual). 【Results】 An overall observation at seven selected sites of the brain indicated that the apoptotic rate induced by orally administrated imidacloprid at sublethal dose (9.90 ng/individual) increased with treatment time and showed significant differences between the treatment group and the control group at 9 and 12 d after treatment (P<0.01). The proportion of Caspase-1 positive cells also increased with the treatment time, and significant differences were observed at 3, 6, 9 and 12 d after treatment (P<0.01). The apoptotic rate and the proportion of Caspase-1 positive cells changed in a timedependent manner, and both showed a positive correlation. The observed ultrastructure of apoptotic nerve cells demonstrated important characteristics of apoptosis and autophagy including cell shrinkage, chromatin condensation, presence of apoptotic bodies and autophagosomes, and swollen mitochondria, some of which were enclosed by autophagic vacuoles and mitophagy occurred. 【Conclusion】 These findings suggest that at the cellular level, a sublethal dose of imidacloprid can induce nerve cell apoptosis in the brain of adult worker honeybees, and the apoptotic pathway is related to Caspase-1 and autophagy. This study provides a useful theoretical basis for estimating the harm of pesticides on non-target organisms.

Key words: Apis mellifera ligustica, neonicotinoid, imidacloprid, apoptosis, autophagy, Caspase-dependent type, ultrastructure