Differential expression of olfactory genes in the southern house mosquito and insights into unique odorant receptor gene isoforms
Differential expression of olfactory genes in the southern house mosquito and insights into unique odorant receptor gene isoforms
复制标题
DOI：
10.1073/pnas.1316059110
复制
发表时间：
2013-11-12
影响因子：
11.1
通讯作者：
Ueira-Vieira, Carlos
中科院分区：
文献类型：
--
作者：
Leal, Walter S.;Choo, Young-Moo;Ueira-Vieira, Carlos
致倦库蚊（Culex quinquefasciatus）是南方的一种蚊子，它的嗅觉系统是迄今为止研究过的所有蚊子中最敏锐和最折衷的。在这里，我们使用Illumina测序来鉴定主要在触角（蚊子的主要嗅觉器官）中表达的嗅觉基因。由高质量文库生成的修剪读段中，不到50%与转录本对齐，但其中约70%与基因组对齐。通过对一组基因进行定量实时PCR验证的差异表达分析表明，48个气味结合蛋白基因中约有一半富集在触角中，另一半主要在腿中表达。类似的模式，观察到与化学感觉蛋白，“加C”气味结合蛋白，感觉神经元膜蛋白。多达43个离子型受体的转录本在雌性触角中富集，从而使离子型受体家族成为触角中最大的嗅觉基因，仅次于气味受体（OR）基因。目前已鉴定出多达177个OR基因，其中包括36个独特的转录本。独特的OR基因不同于以前注释的OR在内部序列，剪接变体，和延长的N或C末端。通过克隆和功能表达验证了以前未知的转录本之一。当用大量生理学相关化合物激发时，CquiOR 95 b以剂量依赖性方式对2-苯乙酸乙酯（其被证明能驱除库蚊）和香茅醛（一种已知的驱虫剂）产生响应。这项转录组研究鉴定了关键分子组分和南方蚊子的驱避剂。
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.