摘要
【目的】miR-124与神经系统发育相关,在多细胞动物脑中特异表达。本研究拟验证miR-124是否在西方蜜蜂3种蜂型的脑中特异表达以及在3种蜂型当中表达差异与功能分析,为西方蜜蜂中miR-124功能研究奠定基础。【方法】设计茎-环状引物反转录miRNA,定量PCR(称为stem-loop RT-qPCR检测法)检测miR-124在西方蜜蜂3种蜂型头部及其他部位表达情况;以邻位相连法构建miR-124系统进化树;利用BioEdit软件对miR-124成熟序列进行同源性分析;查找miR-124在西方蜜蜂当中的靶基因并对靶基因进行功能分析。【结果】miR-124在3种蜂型头部高量表达,其中工蜂最高,雄蜂次之,蜂王最低;miR-124在3种蜂型的其他部位微量表达,相对于3种蜂型头部几乎不表达;多物种中,miR-124同源性最低达到82.6%,最高达到100%;在西方蜜蜂中获得96个miR-124靶基因,45个靶基因获得基因注释,功能归类显示多个miR-124靶基因参与神经发育与调节。【结论】miR-124在3种蜂型头部特异表达,其中在工蜂头部表达最高,雄蜂次之,蜂王最低,而在三型蜂中,由于工蜂的脑最发达,雄蜂的次之,蜂王的最小,推测miR-124与西方蜜蜂3种蜂型的脑发育和调控3种蜂型分工相关。
[Objectives] miR-124 is related to the development of the nervous system and is expressed specifically in the brains of multicellular animals. This study was designed to verify whether expression of miR-124 is restricted to the brain in all three castes of the honeybee Apis mellifera, to analyze the expression of this gene in the three castes, and to provide a foundation for studying the function of miR-124 in the honeybee. [Methods] Real-time Polymerase Chain Reaction （known as stem-loop RT-qPCR assay） was used to detect the expression of miR-124 in the head and other parts of honeybees from each caste using stem-loop primer reverse transcription miRNA. Neighbor joining miR-124 phylogenetic trees were constructed and the software package BioEdit used to analyze the homology of mature miR-124 sequences. A search for the target gene of miR-124 in the honeybee was then conducted and the function of the putative target gene analyzed. [Results] miR-124 was highly expressed in the heads of honeybees from all three castes but was more highly expressed in workers than drones and least expressed in queens. miR-124 expression in body parts other than the head was negligible in all three castes. In many species, the minimum and maximum homology of miR-124 reached 82.6% and 100%, respectively. Ninety-six potential miR-124 target genes were identified in the honeybee, forty-five of which were annotated. Functional classification revealed that multiple miR-124 target genes are involved in neural development and regulation. [Conclusion] miR-124 expression is predominantly confined to the head in all three honeybee castes where it is more highly expressed in workers than drones and least expressed in queens. Worker＆#39;s brains are more developed than those of drones and the brains of queens are the least developed. We suspect that the observed variation in the expression of miR-124 among honeybee castes is related to the level of brain development in each caste, and ultimately to the different roles that each caste plays in the hive.
出处
《应用昆虫学报》
CAS
CSCD
北大核心
2017年第4期568-575,共8页
Chinese Journal of Applied Entomology
基金
国家蜂产业技术体系专项(CARS-45-SYZ 17)
云南省科技厅重点新产品开发计划项目(2015BB010)
云南省农业科学院蚕桑蜜蜂研究所青年创新基金项目(QC2015006)
