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Deep sequencing of the murine olfactory receptor neuron transcriptome.

Kanageswaran N, Demond M, Nagel M, Schreiner BS, Baumgart S, Scholz P, Altmüller J, Becker C, Doerner JF, Conrad H, Oberland S, Wetzel CH, Neuhaus EM, Hatt H, Gisselmann G - PLoS ONE (2015)

Bottom Line: In OE samples, nearly all OR and trace amine-associated receptor (TAAR) genes involved in the perception of volatile amines were detectably expressed.To identify OE-specific genes, we compared olfactory neuron expression profiles with RNA-Seq transcriptome data from different murine tissues.The quantitative and comprehensive transcriptome data provide a virtually complete catalogue of genes expressed in the OE and present a useful tool to uncover candidate genes involved in, for example, olfactory signaling, OR trafficking and recycling, and proliferation.

View Article: PubMed Central - PubMed

Affiliation: Ruhr-University Bochum, Department of Cell Physiology, Bochum, Germany.

ABSTRACT
The ability of animals to sense and differentiate among thousands of odorants relies on a large set of olfactory receptors (OR) and a multitude of accessory proteins within the olfactory epithelium (OE). ORs and related signaling mechanisms have been the subject of intensive studies over the past years, but our knowledge regarding olfactory processing remains limited. The recent development of next generation sequencing (NGS) techniques encouraged us to assess the transcriptome of the murine OE. We analyzed RNA from OEs of female and male adult mice and from fluorescence-activated cell sorting (FACS)-sorted olfactory receptor neurons (ORNs) obtained from transgenic OMP-GFP mice. The Illumina RNA-Seq protocol was utilized to generate up to 86 million reads per transcriptome. In OE samples, nearly all OR and trace amine-associated receptor (TAAR) genes involved in the perception of volatile amines were detectably expressed. Other genes known to participate in olfactory signaling pathways were among the 200 genes with the highest expression levels in the OE. To identify OE-specific genes, we compared olfactory neuron expression profiles with RNA-Seq transcriptome data from different murine tissues. By analyzing different transcript classes, we detected the expression of non-olfactory GPCRs in ORNs and established an expression ranking for GPCRs detected in the OE. We also identified other previously undescribed membrane proteins as potential new players in olfaction. The quantitative and comprehensive transcriptome data provide a virtually complete catalogue of genes expressed in the OE and present a useful tool to uncover candidate genes involved in, for example, olfactory signaling, OR trafficking and recycling, and proliferation.

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OR detection using RNA-Seq: Bar chart showing the percentage of detected OR genes in female and male OE tissue and FACS-sorted ORNs.Percentages were calculated based on the 1,125 OR genes annotated in the Refseq based gene model.
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pone.0113170.g001: OR detection using RNA-Seq: Bar chart showing the percentage of detected OR genes in female and male OE tissue and FACS-sorted ORNs.Percentages were calculated based on the 1,125 OR genes annotated in the Refseq based gene model.

Mentions: To analyze the transcriptome in more detail, we first focused on chemoreceptor gene families starting with OR genes. The high sensitivity of RNA-Seq was demonstrated by the detection of nearly all ORs. Of 1,125 annotated OR genes, we detected 1,075 in female and 1,001 in male CD1 mice and 1,044 in female C57BL/6J mice with FPKMs > 0.1 (Fig. 1, S5 Table, S2 Fig.).


Deep sequencing of the murine olfactory receptor neuron transcriptome.

Kanageswaran N, Demond M, Nagel M, Schreiner BS, Baumgart S, Scholz P, Altmüller J, Becker C, Doerner JF, Conrad H, Oberland S, Wetzel CH, Neuhaus EM, Hatt H, Gisselmann G - PLoS ONE (2015)

OR detection using RNA-Seq: Bar chart showing the percentage of detected OR genes in female and male OE tissue and FACS-sorted ORNs.Percentages were calculated based on the 1,125 OR genes annotated in the Refseq based gene model.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4295871&req=5

pone.0113170.g001: OR detection using RNA-Seq: Bar chart showing the percentage of detected OR genes in female and male OE tissue and FACS-sorted ORNs.Percentages were calculated based on the 1,125 OR genes annotated in the Refseq based gene model.
Mentions: To analyze the transcriptome in more detail, we first focused on chemoreceptor gene families starting with OR genes. The high sensitivity of RNA-Seq was demonstrated by the detection of nearly all ORs. Of 1,125 annotated OR genes, we detected 1,075 in female and 1,001 in male CD1 mice and 1,044 in female C57BL/6J mice with FPKMs > 0.1 (Fig. 1, S5 Table, S2 Fig.).

Bottom Line: In OE samples, nearly all OR and trace amine-associated receptor (TAAR) genes involved in the perception of volatile amines were detectably expressed.To identify OE-specific genes, we compared olfactory neuron expression profiles with RNA-Seq transcriptome data from different murine tissues.The quantitative and comprehensive transcriptome data provide a virtually complete catalogue of genes expressed in the OE and present a useful tool to uncover candidate genes involved in, for example, olfactory signaling, OR trafficking and recycling, and proliferation.

View Article: PubMed Central - PubMed

Affiliation: Ruhr-University Bochum, Department of Cell Physiology, Bochum, Germany.

ABSTRACT
The ability of animals to sense and differentiate among thousands of odorants relies on a large set of olfactory receptors (OR) and a multitude of accessory proteins within the olfactory epithelium (OE). ORs and related signaling mechanisms have been the subject of intensive studies over the past years, but our knowledge regarding olfactory processing remains limited. The recent development of next generation sequencing (NGS) techniques encouraged us to assess the transcriptome of the murine OE. We analyzed RNA from OEs of female and male adult mice and from fluorescence-activated cell sorting (FACS)-sorted olfactory receptor neurons (ORNs) obtained from transgenic OMP-GFP mice. The Illumina RNA-Seq protocol was utilized to generate up to 86 million reads per transcriptome. In OE samples, nearly all OR and trace amine-associated receptor (TAAR) genes involved in the perception of volatile amines were detectably expressed. Other genes known to participate in olfactory signaling pathways were among the 200 genes with the highest expression levels in the OE. To identify OE-specific genes, we compared olfactory neuron expression profiles with RNA-Seq transcriptome data from different murine tissues. By analyzing different transcript classes, we detected the expression of non-olfactory GPCRs in ORNs and established an expression ranking for GPCRs detected in the OE. We also identified other previously undescribed membrane proteins as potential new players in olfaction. The quantitative and comprehensive transcriptome data provide a virtually complete catalogue of genes expressed in the OE and present a useful tool to uncover candidate genes involved in, for example, olfactory signaling, OR trafficking and recycling, and proliferation.

Show MeSH