Dissociated neuronal culture of rats expressing insect odorant receptors as an odorant sensor

人工气味传感器的表现通常不如生物的嗅觉系统。生物气味系统的优异性能是由气味受体的分子识别能力和随之而来的神经信息处理来实现的。为了利用这一优势，我们提出了一种新的混合气味生物传感器，通过将昆虫的嗜离子性气味受体表达到啮齿动物的游离神经元培养物中。这种材料的组合带来了显著的优势，如易于功能表达，延长使用寿命，并能够放大气味受体的弱离子电流。在本研究中，通过脂质体转染，蚕的信息素受体和共受体BmOR1和BmorOrco在神经元培养物中表达。因此，BmOR1和BmorOrco在8%的神经元细胞中共表达，并且两种受体在细胞膜上共定位。在Ca++成像实验中，转染细胞和未转染细胞均发现在出现BOL时钙信号同步增加，且呈剂量依赖性。这些结果证明了所提出的混合气味生物传感器的概念。

Artificial odorant sensors generally perform poorer than olfactory systems in living organisms. The excellent performances of living odorant systems are achieved by the molecular recognition abilities of odorant receptors and the neuronal information processing that follows. To take advantages of this, here we propose a novel hybrid odorant biosensor by means of expressing ionotropic odorant receptors of insects into dissociated neuronal cultures of rodents. This combination of materials brings significant advantages such as easy functional expression, prolonged lifetime, and an ability to amplify the weak ionic currents of odorant receptors. In the present work, pheromone receptors and co-receptors of silkmoth, i. e., BmOR1 and BmorOrco, were expressed in neuronal cultures via liposome transfection. Consequently, BmOR1 and BmorOrco were co-expressed in 8% of neuronal cells, and both receptors were co-localized on a cell membrane. In Ca++imaging experiments, synchronous increase of calcium signals at the presentation of BOL was found in both transfected cells and non-transfected cells in a dose-dependent manner. These results provide the proof-of-concept of the proposed hybrid odorant biosensor.