Bioengineered olfactory sensory neuron-based biosensor for specific odorant detection

It is highly desirable to obtain functional cells with specific olfactory receptors (ORs) for the development of cell-based biosensors towards odorant detection. In this study, we explored the feasibility of bioengineered primary olfactory sensory neurons (OSNs) as sensing elements of biomimetic olfactory-based biosensors, in which light addressable potentiometric sensor (LAPS) was used to monitor bioengineered OSNs membrane potential responses to odorant molecules. An olfactory receptor of C. elegances, ODR-10, as a model receptor, was expressed on the plasma membrane of OSNs by transient transfection. The response profile of bioengineered OSNs to odorant molecules was investigated by analyzing extracellular potential firings features in frequency and time domains. The results indicated that bioengineered OSNs can specifically respond to diacetyl, the natural ligand of ODR-10. In addition, bioengineered OSNs showed different temporal firing patterns in responding to different concentrations of diacetyl. All the results demonstrate that bioengineered OSNs are useful and promising to serve as novel sensing elements of biosensors for specific odorant molecule detection. It is suggested that bioengineering techniques could provide novel approaches for preparing sensitive elements as well as promoting the development of practical applicable olfactory-based biosensors. (C) 2012 Elsevier B.V. All rights reserved.

为了开发用于气味检测的基于细胞的生物传感器，获得具有特定嗅觉受体（ORs）的功能性细胞是非常理想的。在这项研究中，我们探讨了生物工程化的原代嗅觉感觉神经元（OSNs）作为基于仿生嗅觉的生物传感器的传感元件的可行性，其中使用光寻址电位传感器（LAPS）来监测生物工程化的OSNs对气味分子的膜电位响应。秀丽隐杆线虫的一种嗅觉受体ODR - 10作为模型受体，通过瞬时转染在OSNs的质膜上表达。通过分析细胞外电位发放在频率和时间域的特征，研究了生物工程化的OSNs对气味分子的响应特性。结果表明，生物工程化的OSNs能够特异性地对ODR - 10的天然配体双乙酰作出反应。此外，生物工程化的OSNs在对不同浓度的双乙酰作出反应时表现出不同的时间发放模式。所有结果都表明，生物工程化的OSNs作为用于特定气味分子检测的生物传感器的新型传感元件是有用且有前景的。这表明生物工程技术可以为制备敏感元件以及促进实用的基于嗅觉的生物传感器的发展提供新的方法。（C）2012爱思唯尔公司。保留所有权利。