Cell-Based Odorant Sensor Array for Odor Discrimination Based on Insect Odorant Receptors
Cell-Based Odorant Sensor Array for Odor Discrimination Based on Insect Odorant Receptors
复制标题
DOI：
10.1007/s10886-016-0726-7
复制
发表时间：
2016-07-01
影响因子：
2.3
通讯作者：
Kanzaki, Ryohei
中科院分区：
文献类型：
--
作者：
Termtanasombat, Maneerat;Mitsuno, Hidefumi;Kanzaki, Ryohei
生物的嗅觉系统通过识别几种气味受体（ORs）的激活模式，可以准确地辨别多种气味。因此，基于多个ORs的气味传感器阵列的开发提供了模仿生物气味识别机制的可能性。最近，我们基于昆虫or表达的Sf21细胞开发了一种新的高灵敏度和选择性的气味传感器元件，该元件通过显示增强的荧光强度来响应目标气味。本文介绍了一种气味传感器阵列的开发，该阵列由几种表达不同ORs的Sf21细胞系组成。在本研究中，利用聚二甲基硅氧烷膜模板和称为生物相容性膜锚（BAM）的细胞固定化试剂，成功构建了表达Or13a、Or56a、BmOR1和BmOR3的四种细胞系的阵列模式。我们证明BAM可以在不影响其气味感知性能的情况下创建Sf21传感器细胞的清晰模式。我们的传感器阵列显示了对气味混合物和单一气味刺激的气味特异性响应模式，使我们能够将1-辛烯-3-醇、土臭素、蚕蜂素和蚕蜂素的存在可视化为Or13a、Or56a、BmOR1和BmOR3细胞系区域的荧光强度增加。因此，我们成功地开发了一种新的方法来创建一个基于细胞的气味传感器阵列，使我们能够区分多个目标气味。我们的方法可以扩展到一种能够检测大范围环境气味的气味传感器的开发，这可能成为一种有前途的工具，用于各种应用，包括昆虫半化学物质和食品污染的研究。
The olfactory system of living organisms can accurately discriminate numerous odors by recognizing the pattern of activation of several odorant receptors (ORs). Thus, development of an odorant sensor array based on multiple ORs presents the possibility of mimicking biological odor discrimination mechanisms. Recently, we developed novel odorant sensor elements with high sensitivity and selectivity based on insect OR-expressing Sf21 cells that respond to target odorants by displaying increased fluorescence intensity. Here we introduce the development of an odorant sensor array composed of several Sf21 cell lines expressing different ORs. In this study, an array pattern of four cell lines expressing Or13a, Or56a, BmOR1, and BmOR3 was successfully created using a patterned polydimethylsiloxane film template and cell-immobilizing reagents, termed biocompatible anchor for membrane (BAM). We demonstrated that BAM could create a clear pattern of Sf21 sensor cells without impacting their odorant-sensing performance. Our sensor array showed odorant-specific response patterns toward both odorant mixtures and single odorant stimuli, allowing us to visualize the presence of 1-octen-3-ol, geosmin, bombykol, and bombykal as an increased fluorescence intensity in the region of Or13a, Or56a, BmOR1, and BmOR3 cell lines, respectively. Therefore, we successfully developed a new methodology for creating a cell-based odorant sensor array that enables us to discriminate multiple target odorants. Our method might be expanded into the development of an odorant sensor capable of detecting a large range of environmental odorants that might become a promising tool used in various applications including the study of insect semiochemicals and food contamination.