意大利蜜蜂脑神经细胞钙离子浓度测定方法的研究  

The detection method of intracellular Ca^2+ ion concentration of brain neurons of honeybee Apis mellifera ligustica

作　　者：杨俊[1,2] 周玮[1] 周婷[1,2] 王强[1,2] 代平礼[1,2] 吴艳艳[1,2] 

机构地区：[1]中国农业科学院蜜蜂研究所,北京100093 [2]农业部授粉昆虫重点开放实验室,北京100093

出　　处：《应用昆虫学报》2012年第5期1167-1171,共5页Chinese Journal of Applied Entomology

基　　金：国家自然科学基金项目(31072094);"十二五"现代蜂产业技术体系建设专项;中国农业科学院北京畜牧兽医研究所基本科研业务费专项资助

摘　　要：本实验试图建立一套蜜蜂脑神经细胞游离钙离子浓度([Ca2+]i)的体外测定方法。采用Fura-2 acetoxymethyl ester(Fura-2 AM)作为荧光指示剂,对体外培养的意大利蜜蜂Apis mellifera ligustica Spinola脑神经细胞的[Ca2+]i测定方法进行了探索,研究了不同浓度的Fura-2 AM及不同的孵育时间对细胞钙离子测量ratio值的影响。结果测得细胞ratio值随Fura-2AM浓度的增大而降低,孵育时间也会影响ratio值,综合比较得到最佳孵育时间以及最佳染料浓度,并在此基础上制订了一套意大利蜜蜂蜜蜂脑神经细胞钙离子浓度的测定方法,这对进一步建立以蜜蜂神经细胞钙离子浓度变化作为衡量环境有毒物质对蜜蜂的风险研究具有重要意义。A prototype in vitro method to measure intracellular free calcium concentration （ [ Ca2 ＋] i） in honeybee brain cells was developed and tested. Fura-2 AM was used as a florescence indicator to measure intraeellular calcium ion concentration ratio values and incubation time ratio was also recorded and analyzed using META FLOUR software. The results revealed that cell ratio values decreased among groups with increased concentration of Fura-2 AM during eell incubation, and there were highly significant differences between groups （P 〈 0. 01 ）. Similarly, it was clear that incubation time can potentially affect ratio values ; for example only 10 mins incubation resulted in ratio values significantly lower than the other treatments.

关 键 词：蜜蜂 脑神经细胞 FURA-2 AM 钙离子浓度 测量 

分 类 号：S891[农业科学—特种经济动物饲养]