Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase
Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase
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DOI：
10.3791/59446
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发表时间：
2019-04-01
影响因子：
1.2
通讯作者：
Matsunami, Hiroaki
中科院分区：
文献类型：
--
作者：
de March, Claire A.;Fukutani, Yosuke;Matsunami, Hiroaki
嗅觉感知开始于气味与嗅觉感觉神经元（OSN）表达的气味受体（OR）的相互作用。气味识别遵循组合编码方案，其中一个OR可以被一组气味剂激活，并且一种气味剂可以激活OR的组合。通过这种组合编码，生物体可以检测和区分无数的挥发性气味分子。因此，给定浓度下的气味可以通过OR的激活模式来描述，这对每种气味都是特异性的。从这个意义上说，破解大脑用来感知气味的机制需要理解气味-OR相互作用。这就是为什么嗅觉界致力于“去嗅化”这些受体。用于鉴定气味剂-OR相互作用的常规体外系统已经利用了将细胞培养基与气味剂一起孵育，这与通过蒸汽气味剂在与OR相互作用之前溶解到鼻粘膜中的气味的自然检测不同。在这里，我们描述了一种新的方法，允许实时监测或激活通过气相添味剂。我们的方法依赖于使用Glosensor测定通过发光测量cAMP释放。它弥补了目前在体内和体外方法之间的差距，并为仿生挥发性化学传感器提供了基础。
Olfactory perception begins with the interaction of odorants with odorant receptors (OR) expressed by olfactory sensory neurons (OSN). Odor recognition follows a combinatorial coding scheme, where one OR can be activated by a set of odorants and one odorant can activate a combination of ORs. Through such combinatorial coding, organisms can detect and discriminate between a myriad of volatile odor molecules. Thus, an odor at a given concentration can be described by an activation pattern of ORs, which is specific to each odor. In that sense, cracking the mechanisms that the brain uses to perceive odor requires the understanding odorant-OR interactions. This is why the olfaction community is committed to "de-orphanize" these receptors. Conventional in vitro systems used to identify odorant-OR interactions have utilized incubating cell media with odorant, which is distinct from the natural detection of odors via vapor odorants dissolution into nasal mucosa before interacting with ORs. Here, we describe a new method that allows for real-time monitoring of OR activation via vapor-phase odorants. Our method relies on measuring cAMP release by luminescence using the Glosensor assay. It bridges current gaps between in vivo and in vitro approaches and provides a basis for a biomimetic volatile chemical sensor.