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. 2013 Nov 12;110(46):18704-9.
doi: 10.1073/pnas.1316059110. Epub 2013 Oct 28.

Differential expression of olfactory genes in the southern house mosquito and insights into unique odorant receptor gene isoforms

Affiliations

Differential expression of olfactory genes in the southern house mosquito and insights into unique odorant receptor gene isoforms

Walter S Leal et al. Proc Natl Acad Sci U S A. .

Abstract

The southern house mosquito, Culex quinquefasciatus, has one of the most acute and eclectic olfactory systems of all mosquito species hitherto studied. Here, we used Illumina sequencing to identify olfactory genes expressed predominantly in antenna, mosquito's main olfactory organ. Less than 50% of the trimmed reads generated by high-quality libraries aligned to a transcript, but approximately 70% of them aligned to the genome. Differential expression analysis, which was validated by quantitative real-time PCR on a subset of genes, showed that approximately half of the 48 odorant-binding protein genes were enriched in antennae, with the other half being predominantly expressed in legs. Similar patterns were observed with chemosensory proteins, "plus-C" odorant-binding proteins, and sensory neuron membrane proteins. Transcripts for as many as 43 ionotropic receptors were enriched in female antennae, thus making the ionotropic receptor family the largest of antennae-rich olfactory genes, second only to odorant receptor (OR) genes. As many as 177 OR genes have been identified, including 36 unique transcripts. The unique OR genes differed from previously annotated ORs in internal sequences, splice variants, and extended N or C terminus. One of the previously unknown transcripts was validated by cloning and functional expression. When challenged with a large panel of physiologically relevant compounds, CquiOR95b responded in a dose-dependent manner to ethyl 2-phenylacteate, which was demonstrated to repel Culex mosquitoes, and secondarily to citronellal, a known insect repellent. This transcriptome study led to identification of key molecular components and a repellent for the southern house mosquito.

Keywords: RNA-Seq; citronellal polymorphism isoforms.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Differential expression of currently annotated odorant receptors (141 CquiOR genes). One hundred and seven CquiOR genes were significantly expressed in antennae, whereas 8 genes were enriched in legs, and transcripts for 3 genes were not detected.
Fig. 2.
Fig. 2.
Transcript levels (FPKM) for unique CquiORs. Bars in blue are for unique isoforms of currently annotated OR genes (green) and those in red represent unique CquiOR genes. Largely, the transcript levels in legs (Dataset S5) were low and omitted for clarity.
Fig. 3.
Fig. 3.
DEET-induced and ethyl 2-phenylacetate-induced repellency of mosquitoes responding to physical stimuli. Combined results of 2 two-choice assays comparing solvent with DEET, and solvent with the best ligand for CquiOR95b, ethyl 2-phenylacetate (n = 5–6).

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