Research Center for Advanced Science and Technology, The University of Tokyo, Komaba 4-6-1, Tokyo 153-8904, Japan.
Analyst. 2012 Aug 7;137(15):3452-8. doi: 10.1039/c2an35058k. Epub 2012 Jun 20.
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.
人工气味传感器的性能通常不如生物体内的嗅觉系统。生物气味系统之所以具有优异的性能，是因为其气味受体具有分子识别能力，以及紧随其后的神经元信息处理能力。为了利用这一点，我们通过在啮齿动物分离神经元培养物中表达昆虫的离子型气味受体，提出了一种新型混合气味生物传感器。这种材料的结合具有易于功能表达、寿命延长和放大气味受体弱离子电流的能力。在本工作中，通过脂质体转染，在神经元培养物中表达了家蚕的信息素受体和共受体，即 BmOR1 和 BmorOrco。结果表明，BmOR1 和 BmorOrco 在 8%的神经元细胞中共表达，并且这两种受体都定位于细胞膜上。在 Ca++成像实验中，在 BOL 呈现时，转染细胞和未转染细胞中的钙信号呈剂量依赖性同步增加。这些结果为所提出的混合气味生物传感器提供了概念验证。
