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PLoS One. 2014 May 6;9(5):e96333. doi: 10.1371/journal.pone.0096333. eCollection 2014.

Profiling of olfactory receptor gene expression in whole human olfactory mucosa.

Author information

1
Department of Otorhinolaryngology, Erasme University Hospital, Brussels, Belgium.
2
ChemCom S.A., Brussels, Belgium.
3
Institute of Interdisciplinary Research in human and molecular Biology, Free University of Brussels, Brussels, Belgium.
4
Laboratory of Pathophysiological and Nutritional Biochemistry, Department of Biochemistry, Free University of Brussels, Brussels, Belgium.

Abstract

Olfactory perception is mediated by a large array of olfactory receptor genes. The human genome contains 851 olfactory receptor gene loci. More than 50% of the loci are annotated as nonfunctional due to frame-disrupting mutations. Furthermore haplotypic missense alleles can be nonfunctional resulting from substitution of key amino acids governing protein folding or interactions with signal transduction components. Beyond their role in odor recognition, functional olfactory receptors are also required for a proper targeting of olfactory neuron axons to their corresponding glomeruli in the olfactory bulb. Therefore, we anticipate that profiling of olfactory receptor gene expression in whole human olfactory mucosa and analysis in the human population of their expression should provide an opportunity to select the frequently expressed and potentially functional olfactory receptors in view of a systematic deorphanization. To address this issue, we designed a TaqMan Low Density Array (Applied Biosystems), containing probes for 356 predicted human olfactory receptor loci to investigate their expression in whole human olfactory mucosa tissues from 26 individuals (13 women, 13 men; aged from 39 to 81 years, with an average of 67±11 years for women and 63±12 years for men). Total RNA isolation, DNase treatment, RNA integrity evaluation and reverse transcription were performed for these 26 samples. Then 384 targeted genes (including endogenous control genes and reference genes specifically expressed in olfactory epithelium for normalization purpose) were analyzed using the same real-time reverse transcription PCR platform. On average, the expression of 273 human olfactory receptor genes was observed in the 26 selected whole human olfactory mucosa analyzed, of which 90 were expressed in all 26 individuals. Most of the olfactory receptors deorphanized to date on the basis of sensitivity to known odorant molecules, which are described in the literature, were found in the expressed olfactory receptors gene set.

PMID:
24800820
PMCID:
PMC4011832
DOI:
10.1371/journal.pone.0096333
[Indexed for MEDLINE]
Free PMC Article

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