Biosensor National Special Laboratory, Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China.
Biosens Bioelectron. 2013 Feb 15;40(1):401-6. doi: 10.1016/j.bios.2012.08.035. Epub 2012 Sep 12.
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.
获得具有特定嗅觉受体 (OR) 的功能细胞对于开发基于细胞的生物传感器以进行气味检测是非常可取的。在这项研究中,我们探索了生物工程化的初级嗅觉感觉神经元 (OSN) 作为仿生嗅觉生物传感器的传感元件的可行性,其中光寻址电位传感器 (LAPS) 用于监测生物工程化 OSN 对气味分子的膜电位响应。通过瞬时转染,将秀丽隐杆线虫的嗅觉受体 ODR-10 作为模型受体表达在 OSN 的质膜上。通过分析频率和时域中的细胞外电势激发特征,研究了生物工程化 OSN 对气味分子的响应特征。结果表明,生物工程化 OSN 可以特异性地响应二乙酰,这是 ODR-10 的天然配体。此外,生物工程化 OSN 在响应不同浓度的二乙酰时表现出不同的时间发射模式。所有结果表明,生物工程化 OSN 可用作生物传感器的新型传感元件,用于特定气味分子的检测。建议生物工程技术可以为制备敏感元件提供新方法,并促进实用的基于嗅觉的生物传感器的发展。