遗传 ›› 2018, Vol. 40 ›› Issue (2): 155-161.doi: 10.16288/j.yczz.17-096

• 研究报告 • 上一篇    下一篇

中华蜜蜂酪胺受体基因克隆及表达分析

张丽珍(),张永(),胡景华,王子龙,曾志将   

  1. 江西农业大学蜜蜂研究所,南昌 330045
  • 收稿日期:2017-03-21 修回日期:2017-12-12 出版日期:2018-02-20 发布日期:2018-01-11
  • 作者简介:作者简介: 张丽珍,博士,实验师,研究方向:蜜蜂行为及分子生物学。E-mail: zlzcg@126.com|张永,硕士研究生,专业方向:蜜蜂分子生物学。E-mail: 502315963@qq.com|通讯作者: 曾志将,博士,教授,博士生导师,研究方向:蜜蜂生物学。E-mail: bees1965@sina.com|张丽珍和张永为并列第一作者。
  • 基金资助:
    国家自然科学基金项目(31260524,31602015,31402147);国家蜂产业技术体系项目资助(CARS-45-KXJ12)

Molecular cloning and expression analysis of the tyramine
receptor genes in Apis cerana cerana

Zhang Lizhen(),Zhang Yong(),Hu Jinghua,Wang Zilong,Zeng Zhijiang   

  1. Honeybee Research Institute, Jiangxi Agricultural University, Nanchang 330045, China
  • Received:2017-03-21 Revised:2017-12-12 Online:2018-02-20 Published:2018-01-11
  • Supported by:
    the National Natural Science Foundation of China(31260524,31602015,31402147);the Earmarked Fund for the China Agriculture Research System(CARS-45-KXJ12)

摘要:

酪胺(tyramine)属于生物多聚胺类,是昆虫中枢神经系统内重要的神经递质、神经调质和神经激素,参与调控昆虫的多种行为和生理过程,如酪胺受体基因参与调控动物的学习与记忆。本研究首次克隆获得中华蜜蜂(Apis cerana cerana)酪胺受体基因Actyr1Actyr2的全长cDNA序列,利用qRT-PCR方法鉴定了Actyr1Actyr2在中华蜜蜂不同组织器官中的表达谱,采用地高辛原位杂交技术对Actyr1Actyr2在大脑中的表达进行了定位。中华蜜蜂Actyr1Actyr2的cDNA全长序列分别为1241 bp(GenBank登录号:KC814693)和1270 bp(GenBank登录号:KC814694),分别编码297、399个氨基酸残基。qRT-PCR分析结果表明,Actyr1Actyr2在不同组织中的表达量为头部最高,其次是腹部表皮,触角和胸部肌肉的表达量最低,并且头部的表达量显著高于其他组织的表达量;原位杂交结果显示,Acytr1Actyr2在中华蜜蜂大脑蘑菇体的凯尼恩细胞、触角叶周围的细胞处均有较强阳性着色。这些研究表明,Acytr1Actyr2基因可能参与了蜜蜂的学习记忆,并且在相同的细胞中互相作用,共同调控蜜蜂的生物学功能。

关键词: 中华蜜蜂, 酪胺受体基因, 分子克隆, 表达分析

Abstract:

Tyramine is a biological polyamine, which serves important functions as neurotransmitters, neuromodulators and neurohormone of the central nervous system. It participates in the regulation of various behavior and physiological processes in insects. For example, tyramine and its receptor genes are involved in the regulation of learning and memory in the animals. In this study, the full-length cDNA sequences of the tyramine receptor genes (Actyr1 and Actyr2) of the Chinese honeybee, Apis cerana cerana, were cloned and sequenced for the first time. Their expression patterns were examined in different tissues by qRT-PCR and localized in the head by in situ hybridization with digoxigenin (DIG)-labeled RNA probes. The full-length cDNAs of Actyr1 and Actyr2 are 1241 bp (GenBank accession no. KC814693) and 1270 bp (GenBank accession no.KC814693) in length and encode 297 amino acids and 399 amino acids, respectively. qRT-PCR results showed that the expression levels of both Actyr1 and Actyr2 were the highest in the head, followed by the abdomen, then the antennae and the lowest in the thorax. The expression level in the head was significantly higher than that in other tissues. Moreover, in situ hybridization showed that the expression of Actyr1 and Actyr2 genes were mainly localized to the Kenyon cells of the mushroom bodies and cells around the antennal lobes. These observations suggest that some interactions between these two genes in certain cells could be important in regulating various biological functions, such as learning and memory, in the honeybee.

Key words: Apis cerana cerana, tyramine receptor genes, molecular cloning, expression analysis