Department of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA.
J Insect Physiol. 2013 Sep;59(9):961-6. doi: 10.1016/j.jinsphys.2013.07.004. Epub 2013 Jul 19.
The Southern house mosquito Culex quinquefasciatus has the largest repertoire of odorant receptors (ORs) of all mosquitoes and dipteran species whose genomes have been sequenced to date. Previously, we have identified and de-orphanized two ORs expressed in female antennae, CquiOR2 and CquiOR10, which are sensitive to oviposition attractants. In view of a new nomenclature for the Culex genome (VectorBase) we renamed these ORs as CquiOR21 (formerly CquiOR10) and CquiOR121 (CquiOR2). In addition, we selected ORs from six different phylogenetic groups for deorphanization. We cloned four of them by using cDNA from female antennae as a template. Attempts to clone CquiOR87 and CquiOR110 were unsuccessful either because they are pseudogenes or are not expressed in adult female antennae, the main olfactory tissue. By contrast, CquiOR1, CquiOR44, CquiOR73, and CquiOR161 were highly expressed in female antennae. To de-orphanize these ORs, we employed the Xenopus oocyte recording system. CquiORx-CquiOrco-expressed oocytes were challenged with a panel of 90 compounds, including known oviposition attractants, human and vertebrate host odorants, plant kairomones, and naturally occurring repellents. While CquiOR161 did not respond to any test compound in two different laboratories, CquiOR1 showed the features of a generic OR, with strong responses to 1-octen-3-ol and other ligands. CquiOR44 and CquiOR73 showed preference to plant-derived terpenoids and phenolic compounds, respectively. While fenchone was the best ligand for the former, 3,5-dimethylphenol elicited the strongest responses in the latter. The newly de-orphanized ORs may be involved in reception of plant kairomones and/or natural repellents.
南方家蚊库蚊(Culex quinquefasciatus)拥有所有测序基因组的蚊子和双翅目物种中最大的气味受体(OR)基因库。之前,我们已经鉴定并去除了两个在雌性触角中表达的 OR 的孤儿基因,CquiOR2 和 CquiOR10,它们对产卵引诱剂敏感。鉴于 Culex 基因组的新命名法(VectorBase),我们将这两个 OR 重新命名为 CquiOR21(以前称为 CquiOR10)和 CquiOR121(CquiOR2)。此外,我们从六个不同的系统发育群中选择 OR 进行去孤儿化。我们使用雌性触角的 cDNA 作为模板,克隆了其中的四个。尝试克隆 CquiOR87 和 CquiOR110 都没有成功,要么是因为它们是假基因,要么是因为它们在成年雌性触角(主要嗅觉组织)中不表达。相比之下,CquiOR1、CquiOR44、CquiOR73 和 CquiOR161 在雌性触角中高度表达。为了对这些 OR 进行去孤儿化,我们使用了非洲爪蟾卵母细胞记录系统。用一组 90 种化合物对 CquiORx-CquiOrco 表达的卵母细胞进行了挑战,包括已知的产卵引诱剂、人类和脊椎动物宿主气味、植物信息素和天然驱避剂。虽然 CquiOR161 在两个不同的实验室中对任何测试化合物都没有反应,但 CquiOR1 表现出通用 OR 的特征,对 1-辛烯-3-醇和其他配体有强烈的反应。CquiOR44 和 CquiOR73 分别对植物衍生的萜类化合物和酚类化合物表现出偏好。对于前者,fenchone 是最好的配体,而 3,5-二甲基苯酚在后者中引起最强的反应。新去孤儿化的 OR 可能参与植物信息素和/或天然驱避剂的接收。