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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.We also identified other previously undescribed membrane proteins as potential new players in olfaction.

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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In situ hybridization for nGPCRs mRNA.A: Adipor1 (adiponectin receptor 1) B: Gpr178; C: Gpr155; D: Gpr177 (aka Wls); E: Ptgdr (prostaglandin D receptor). All transcripts were detected in the mature ORN cell layer as predicted by the expression levels observed in sorted ORNs. Scale bar = 30 µm.
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pone.0113170.g007: In situ hybridization for nGPCRs mRNA.A: Adipor1 (adiponectin receptor 1) B: Gpr178; C: Gpr155; D: Gpr177 (aka Wls); E: Ptgdr (prostaglandin D receptor). All transcripts were detected in the mature ORN cell layer as predicted by the expression levels observed in sorted ORNs. Scale bar = 30 µm.

Mentions: Finally, we constructed a quantitative ranking of the expression of all GPCRs detected in the OE and ORNs (S7 Table). We selected some of these new nGPCRs and verified their expression in ORNs with in situ hybridization (Fig. 7). All selected probes produced signals in the mature ORN layer as predicted by the FPKM values from ORNs. Additionally, we tested the most highly expressed GPCR, Adipor1 and two other identified GPCRs with RT-PCR and determined their relative expression levels (Fig. 8). As expected, we found that Adipor1 was most highly expressed followed by a lesser extent for Gpr177. These results match our RNA-Seq and in situ hybridization results.


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)

In situ hybridization for nGPCRs mRNA.A: Adipor1 (adiponectin receptor 1) B: Gpr178; C: Gpr155; D: Gpr177 (aka Wls); E: Ptgdr (prostaglandin D receptor). All transcripts were detected in the mature ORN cell layer as predicted by the expression levels observed in sorted ORNs. Scale bar = 30 µm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0113170.g007: In situ hybridization for nGPCRs mRNA.A: Adipor1 (adiponectin receptor 1) B: Gpr178; C: Gpr155; D: Gpr177 (aka Wls); E: Ptgdr (prostaglandin D receptor). All transcripts were detected in the mature ORN cell layer as predicted by the expression levels observed in sorted ORNs. Scale bar = 30 µm.
Mentions: Finally, we constructed a quantitative ranking of the expression of all GPCRs detected in the OE and ORNs (S7 Table). We selected some of these new nGPCRs and verified their expression in ORNs with in situ hybridization (Fig. 7). All selected probes produced signals in the mature ORN layer as predicted by the FPKM values from ORNs. Additionally, we tested the most highly expressed GPCR, Adipor1 and two other identified GPCRs with RT-PCR and determined their relative expression levels (Fig. 8). As expected, we found that Adipor1 was most highly expressed followed by a lesser extent for Gpr177. These results match our RNA-Seq and in situ hybridization results.

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.We also identified other previously undescribed membrane proteins as potential new players in olfaction.

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
Related in: MedlinePlus