heterologous-expression-functional-analysis-aedes-aegypti-odorant

Heterologous Expression and Functional Analysis of <em>Aedes aegypti</em> Odorant Receptors to Human Odors in <em>Xenopus</em> Oocytes

The mosquito Aedes aegypti (Linnaeus), a vector of many important human diseases including yellow fever, dengue fever and Zika fever, shows a strong ...

Auburn University

University of California, San Francisco

Quantitative characterization of magnetic particles is useful for analysis and separation of labeled cells and magnetic particles. A particle velocimeter is used to directly measure the magnetophoretic mobility, size, and other parameters of magnetic particle suspensions. The instrument provides quantitative video analysis of particles and their motion. The trajectories of magnetic particles in an isodynamic magnetic field are recorded using a high-definition camera/microscope system for image collection. Image analysis software then converts the image data to the parameters of interest. The distribution of magnetophoretic mobility is determined by combining fast image analysis with velocimetry measurements. Particle size distributions have been characterized to provide a better understanding of sample quality. The results have been used in the development and operation of analyzer protocols for counting particle concentrations accurately and measuring magnetic susceptibility and size for simultaneous display for routine application to particle suspensions and magnetically labeled biological cells. © 2016 International Society for Advancement of Cytometry.

Magnetic particle characterization-magnetophoretic mobility and particle size.

As one of the most notorious ectoparasites, bed bugs rely heavily on human or animal blood sources for survival, mating and reproduction. Chemoreception, mediated by the odorant receptors on the membrane of olfactory sensory neurons, plays a vital role in their host seeking and risk aversion processes. We investigated the responses of odorant receptors to a large spectrum of semiochemicals, including human odorants and plant-released volatiles and found that strong responses were sparse; aldehydes/ketones were the most efficient stimuli, while carboxylic acids and aliphatics/aromatics were comparatively less effective in eliciting responses from bed bug odorant receptors. In bed bugs, both the odorant identity and concentrations play important roles in determining the strength of these responses. The odor space constructed based on the responses from all the odorant receptors tested revealed that odorants within the same chemical group are widely dispersed while odorants from different groups are intermingled, suggesting the complexity of odorant encoding in the bed bug odorant receptors. This study provides a comprehensive picture of the olfactory coding mechanisms of bed bugs that will ultimately contribute to the design and development of novel olfactory-based strategies to reduce both the biting nuisance and disease transmission from bed bugs.

Molecular Basis of Olfactory Chemoreception in the Common Bed Bug, Cimex lectularius.

The yellow fever mosquito, Aedes aegypti, is a vector of many human diseases such as yellow fever, dengue fever, and Zika. As insecticide resistance has been widely reported, chemical repellents have been adopted as alternative options for mosquito and mosquito-borne disease control. This study characterized the responses of olfactory receptor neurons (ORNs) in different types of antennal olfactory sensilla in Ae. aegypti to 48 chemicals that exhibited repellent activity in various insect species. Both excitatory and inhibitory responses were observed from ORNs in response to these chemicals and differential tuning properties were also observed among ORNs. Remarkable excitatory responses were recorded from the ORNs in sensilla SST1, SST2, SBTI, SBTII, and LST2, while inhibitory activities were detected from a neuron in sensillum SST2 in response to several terpene/terpenoid compounds. Moreover, the temporal dynamics of neuronal responses were found to be compound-specific and concentration-dependent. Hierarchical cluster analysis and principal component analysis of the response to each compound across ORNs in seven types of olfactory sensilla in Ae. aegypti revealed that odor reception depended not only on chemical class but also specific chemical structure. Results of this study give new insights into the sensory physiology of Aedes mosquitoes to the chemical repellents and should contribute to the development of new repellent reagents for human protection.

Neuronal Responses of Antennal Olfactory Sensilla to Insect Chemical Repellents in the Yellow Fever Mosquito, Aedes aegypti.

How chemical signals are integrated at the peripheral sensory system of insects is still an enigma. Here we show that when coexpressed with Orco in Xenopus oocytes, an odorant receptor from the southern house mosquito, CquiOR32, generated inward (regular) currents when challenged with cyclohexanone and methyl salicylate, whereas eucalyptol and fenchone elicited inhibitory (upward) currents. Responses of CquiOR32-CquiOrco-expressing oocytes to odorants were reduced in a dose-dependent fashion by coapplication of inhibitors. This intrareceptor inhibition was also manifested in vivo in fruit flies expressing the mosquito receptor CquiOR32, as well in neurons on the antennae of the southern house mosquito. Likewise, an orthologue from the yellow fever mosquito, AaegOR71, showed intrareceptor inhibition in the Xenopus oocyte recording system and corresponding inhibition in antennal neurons. Inhibition was also manifested in mosquito behavior. Blood-seeking females were repelled by methyl salicylate, but repellence was significantly reduced when methyl salicylate was coapplied with eucalyptol.

Odorant Inhibition in Mosquito Olfaction.

Insects use their olfactory systems to obtain chemical information on mating partners, oviposition sites and food. The yellow fever mosquito Aedes aegypti, an important vector of human infectious diseases, shows strong preference for human blood meals. This study investigated the chemical basis of host detection by characterizing the neuronal responses of antennal olfactory sensilla of female Ae. aegypti to 103 compounds from human skin emanations. The effect of blood feeding on the responses of olfactory sensilla to these odorants was examined as well. Sensilla SBTII, GP, and three functional subtypes of SST (SST1, SST2, and SST3) responded to most of the compounds tested. Olfactory receptor neurons (ORNs) 'A' and 'B' in the trichoid sensilla, either activated or inhibited, were involved in the odour coding process. Compounds from different chemical classes elicited responses with different temporal structures and different response patterns across the olfactory sensilla. Except for their increased responses to several odorants, blood-fed mosquitoes generally evoked reduced responses to specific aldehydes, alcohols, aliphatics/aromatics, ketones, and amines through the SST1, SST2, SBTI, SBTII and GP sensilla. The odorants eliciting diminished responses in female mosquitoes after blood feeding may be important in Ae. aegypti host-seeking activity and thus can be candidates for mosquito attractants in the process of this disease vector management.

Zhou Chen

Department of Entomology and Plant Pathology

Auburn University

Heterologous Expression and Functional Analysis of Aedes aegypti Odorant Receptors to Human Odors in Xenopus Oocytes

Zhou Chen

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Entomology and Plant Pathology

Auburn University

Heterologous Expression and Functional Analysis of Aedes aegypti Odorant Receptors to Human Odors in Xenopus Oocytes

Department of Entomology and Plant Pathology