Dissociated neuronal culture expressing ionotropic odorant receptors as a hybrid odorant biosensor – proof-of-concept study –
Dissociated neuronal culture expressing ionotropic odorant receptors as a hybrid odorant biosensor – proof-of-concept study –
复制标题
表达离子型气味受体的分离神经培养物作为混合气味生物传感器 – 概念验证研究 –
作者：
Chib V;Yun K;Takahashi H;Shimojo S;Norio Tanada
人工气味传感器通常比生物体中的嗅觉系统表现差。活体气味系统的优异性能是通过气味受体的分子识别能力和随之而来的神经元信息处理来实现的。为了利用这一点，在这里，我们提出了一种新的混合气味生物传感器，通过表达昆虫的离子型气味受体到啮齿动物的解离神经元培养物。这种材料的组合带来了显着的优势，例如易于功能表达、延长寿命以及放大气味受体弱离子电流的能力。本工作研究了家蚕的信息素受体和辅助受体，通过脂质体转染在神经元培养物中表达BmOR 1和BmorOrco。因此，BmOR 1和BmorOrco在8%的神经元细胞中共表达，并且两种受体共定位于细胞膜上。在Ca++成像实验中，在BOL呈现时，在转染细胞和未转染细胞中均发现以剂量依赖性方式的钙信号同步增加。这些结果提供了所提出的混合气味生物传感器的概念验证。
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.