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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Functional identification and reconstitution of an odorant receptor in single olfactory neurons
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Functional identification and reconstitution of an odorant receptor in single olfactory neurons

机译：嗅觉神经元中气味受体的功能鉴定和重构

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The olfactory system is remarkable in its capacity to discriminate a wide range of odorants through a series of transduction events initiated in olfactory receptor neurons. Each olfactory neuron is expected to express only a single odorant receptor gene that belongs to the G protein coupled receptor family. The ligand-receptor intcraction, however, has not been clearly characterized. This study dem- onstrates the functional identification of olfactory receptor(s) for specific odorant(s) from single olfactory neurons by a combination of Caz+-imaging and reverse transcription- coupled PCR analysis. First, a candidate odorant receptor was cloned from a single tissue-printed olfactory neuron that displayed odorant-induced Caz+ increase. Next, recombinant adenovirus-mediated expression of the isolated receptor gene was established in the olfactory epithelium by using grcen fluorescent protein as a marker. The infected neurons elicited cxternal Caz+ entry when exposed to the odorant that origi- nally was used to identify the receptor gene. Experiments performed to determine ligand specificity revealed that the odorant receptor recognized specific structural motifs within odorant molecules. The odorant receptor-mediated signal transduction appears to be reconstituted by this two-step approach: the receptor screening for given odorant(s) from single neurons and the functional expression or the receptor via recombinant adenovirus. The present approach should enable us to examine not only ligand specificity of an odorant receptor but also receptor specificity and diversity for a part
机译：嗅觉系统在嗅觉受体神经元中引发的一系列转导事件中，能够区分各种气味的能力非常出色。期望每个嗅觉神经元仅表达属于G蛋白偶联受体家族的单个气味受体基因。然而，配体-受体整合尚不明确。这项研究通过结合Caz +成像和逆转录偶联PCR分析，证明了单个嗅觉神经元中特定气味的嗅觉受体的功能鉴定。首先，从单个组织印刷的嗅觉神经元中克隆出一个候选的气味受体，该嗅觉神经元显示出气味诱导的Caz +增加。接下来，通过使用grcen荧光蛋白作为标记，在嗅觉上皮中建立了分离的受体基因的重组腺病毒介导的表达。当被感染的神经元暴露于最初用来识别受体基因的气味时，会引起Cazternal Caz +进入。确定配体特异性的实验表明，气味受体能识别气味分子内的特定结构基序。气味受体介导的信号转导似乎可以通过两步方法进行重构：从单个神经元中筛选给定的气味，然后通过重组腺病毒筛选功能性表达或受体。本方法应使我们不仅可以检查气味受体的配体特异性，还可以检查零件的受体特异性和多样性

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