Analyzing responses of mouse olfactory sensory neurons using the air-phase electroolfactogram recording.
Analyzing responses of mouse olfactory sensory neurons using the air-phase electroolfactogram recording.
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DOI：
10.3791/1850
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发表时间：
2010-03-02
期刊：
影响因子：
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通讯作者：
Zhao, Haiqing
中科院分区：
文献类型：
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作者：
Cygnar, Katherine D;Stephan, Aaron B;Zhao, Haiqing
动物依靠嗅觉进行许多重要的行为，如寻找食物来源，避免捕食者，识别交配和其他社会互动的同种。嗅电图（EOG）是一种信息丰富、操作简便、可靠的在嗅上皮水平检测嗅觉功能的方法。自1956年Ottoson描述青蛙的眼电图以来，眼电图记录已被应用于许多脊椎动物，包括蝾螈、兔子、大鼠、小鼠和人类（Scott和Scott-Johnson，2002，参考文献2）。最近在小鼠遗传修饰方面的进展重新点燃了人们对记录敲除和敲入小鼠嗅觉功能生理特征的EOG的兴趣。眼电图记录已成功地应用于证明嗅觉信号转导成分的中心作用，最近的特点，某些监管机制的贡献OSN响应。气味检测发生在OSN的纤毛上的嗅上皮的表面，其中信号转导级联导致离子通道的打开，产生流入纤毛并使膜去极化的电流。EOG是在气味刺激时在嗅上皮表面细胞外记录的负电位，其由记录场中的个体响应性OSN引起的电位变化的总和产生。因此，EOG的振幅和动力学的比较提供了关于基因修饰和其他实验操作如何影响OSN对气味的响应的分子信号传导的有价值的信息。在这里，我们描述了一个空气相位眼电图记录的准备小鼠嗅鼻甲。简言之，在处死小鼠后，通过沿中线沿着平分头部并去除中隔来暴露嗅鼻甲。然后将鼻甲骨制备物置于记录装置中，并将记录电极置于其中一个内侧鼻甲骨上的嗅上皮表面。参比电极通过缓冲溶液电连接到组织。一股连续的加湿空气流吹过上皮细胞的表面以保持其湿润。气味剂溶液的蒸气被喷入加湿空气流中以刺激上皮。对答复进行记录和数字化，以便进一步分析。
Animals depend on olfaction for many critical behaviors, such as finding food sources, avoiding predators, and identifying conspecifics for mating and other social interactions. The electroolfactogram (EOG) recording is an informative, easy to conduct, and reliable method to assay olfactory function at the level of the olfactory epithelium. Since the 1956 description of the EOG by Ottoson in frogs, the EOG recording has been applied in many vertebrates including salamanders, rabbits, rats, mice, and humans (reviewed by Scott and Scott-Johnson, 2002, ref. 2). The recent advances in genetic modification in mice have rekindled interest in recording the EOG for physiological characterization of olfactory function in knock-out and knock-in mice. EOG recordings have been successfully applied to demonstrate the central role of olfactory signal transduction components, and more recently to characterize the contribution of certain regulatory mechanisms to OSN responses. Odorant detection occurs at the surface of the olfactory epithelium on the cilia of OSNs, where a signal transduction cascade leads to opening of ion channels, generating a current that flows into the cilia and depolarizes the membrane. The EOG is the negative potential recorded extracellularly at the surface of the olfactory epithelium upon odorant stimulation, resulting from a summation of the potential changes caused by individual responsive OSNs in the recording field. Comparison of the amplitude and kinetics of the EOG thus provide valuable information about how genetic modification and other experimental manipulations influence the molecular signaling underlying the OSN response to odor. Here we describe an air-phase EOG recording on a preparation of mouse olfactory turbinates. Briefly, after sacrificing the mouse, the olfactory turbinates are exposed by bisecting the head along the midline and removing the septum. The turbinate preparation is then placed in the recording setup, and a recording electrode is placed at the surface of the olfactory epithelium on one of the medial turbinates. A reference electrode is electrically connected to the tissue through a buffer solution. A continuous stream of humidified air is blown over the surface of the epithelium to keep it moist. The vapor of odorant solutions is puffed into the stream of humidified air to stimulate the epithelium. Responses are recorded and digitized for further analysis.