Functional validation of the carbon dioxide receptor in labial palps of Helicoverpa armigera moths
Functional validation of the carbon dioxide receptor in labial palps of Helicoverpa armigera moths
复制标题
棉铃虫唇须二氧化碳受体的功能验证
DOI：
10.1016/j.ibmb.2016.04.002
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发表时间：
2016-06-01
影响因子：
3.8
通讯作者：
Wang, Chen-Zhu
中科院分区：
文献类型：
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作者：
Ning, Chao;Yang, Ke;Wang, Chen-Zhu
成虫的下唇须上有一个器官，称为下唇须窝器官，专门用来感应二氧化碳。它们利用二氧化碳作为线索来探测健康的植物，并在上面寻找食物或产卵。果蝇和埃及伊蚊的CO2受体的分子基础已被报道，但鳞翅目昆虫CO2受体的分子机制仍不清楚。本研究首先对棉铃虫CO2味觉受体基因HarmGr1、HarmGr2和HarmGr3进行了重新检测，并对其表达模式进行了分析。RT-PCR结果表明，该基因主要表达于黑腹果蝇的下唇须。armigera。因此，我们使用原位杂交定位三个基因在唇须中的表达。我们发现，所有三个基因共表达在相同的细胞的唇须。接下来，我们利用非洲爪蟾卵母细胞表达系统和双电极电压钳记录技术研究了这三个基因的功能。结果表明，只有共表达HarmGr1和HarmGr3或共表达HarmGr1，HarmGr2和HarmGr3的卵母细胞对NaHCO3产生了强烈的反应。最后，我们证实，感觉细胞在女性和男性的唇须显示剂量依赖性反应的CO2刺激，通过使用单感器记录。我们的工作揭示了HarmGr1和HarmGr3是梭子蟹唇须CO2传感器不可缺少的充分条件。armigera。(C)2016爱思唯尔有限公司版权所有
Adult moths possess an organ in their labial palps, the labial-palp pit organ, which is specialized for sensing carbon dioxide (CO2). They use CO2 as a cue to detect healthy plants and find food or lay eggs on them. The molecular bases of the CO2 receptor in Drosophila melanogaster and Aedes aegypti have been reported, but the molecular mechanisms of the CO2 receptor in Lepidoptera remains elusive. In this study, we first re-examined three putative Helicoverpa armigera CO2 gustatory receptor genes (HarmGr1, HarmGr2, and HarmGr3), and then analyzed expression patterns of them. RT-PCR results verified they were predominantly expressed in the labial palps of H. armigera. Thus, we used in situ hybridization to localize the expression of three genes in the labial palps. We found that all three genes were co-expressed in the same cells of the labial palps. Next, we employed the Xenopus laevis oocyte expression system and the two-electrode voltage-clamp recording to study the function of the three genes. Results showed that only oocytes co-expressing HarmGrl and HarmGr3 or co-expressing HarmGr1, HarmGr2 and HarmGr3 gave robust responses to NaHCO3. Finally, we confirmed that the sensory cells in labial palps of both females and males show dose dependent responses to CO2 stimuli by using single sensillum recording. Our work uncovers that HarmGrl and HarmGr3 are indispensable and sufficient for CO2 sensing in labial palps of H. armigera. (C) 2016 Elsevier Ltd. All rights reserved.