| Project/Area Number |
09670121
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | KOBE UNIVERSITY (1998)
The University of Tokyo (1997) |
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Principal Investigator |
TOUHARA Kazushige Kobe University Biosignal Research Center Joshu, バイオシグナル研究センター, 助手 (00280925)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1998: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | olfaction / odorant / receptor / cloning / reconstitution / adenovirus / expression / カルシウム / PCR |
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| Research Abstract |
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 interaction, however, has not been clearly characterized. This study is the first to demonstrate the functional identification of olfactory receptor(s) for specific odorant(s) from single olfactory neurons by a combination of Ca^<2+>-imaging and RT-PCR analysis. First, a candidate odorant receptor was cloned from a single tissue-printed olfactory neuron that displayed odorant-induced Ca^<2+>-increase. Next, recombinant adenovirus-mediated expression of the isolated receptor gene was established in the olfactory epithelium using green fluorescent protein as a marker. The infected neurons elicited external Ca^<2+>-entry when exposed to the odorant that was originally utilized 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 of the receptor via recombinant adenovirus. The present approach will enable us to examine not only ligand specificity of an odorant receptor but also receptor specificity and diversity for a particular odorant of interest.
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