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Insect Biochem Mol Biol

Title:		Functional characterization of odorant receptors from the moth Eriocrania semipurpurella: A comparison of results in the Xenopus oocyte and HEK cell systems
Author(s):		Hou X; Zhang DD; Yuvaraj JK; Corcoran JA; Andersson MN; Lofstedt C;
Address:		"Department of Biology, Lund University, Lund, Sweden. Electronic address: xiaoqing.hou@biol.lu.se. Department of Biology, Lund University, Lund, Sweden. Electronic address: dan-dan.zhang@biol.lu.se. Department of Biology, Lund University, Lund, Sweden. Electronic address: jothi_kumar.yuvaraj@biol.lu.se. Department of Biology, Lund University, Lund, Sweden. Electronic address: jacob.corcoran430@gmail.com. Department of Biology, Lund University, Lund, Sweden. Electronic address: martin_n.andersson@biol.lu.se. Department of Biology, Lund University, Lund, Sweden. Electronic address: christer.lofstedt@biol.lu.se"
Journal Title:		Insect Biochem Mol Biol
Year:		2020
Volume:		20191125
Issue:
Page Number:		103289 -
DOI:		10.1016/j.ibmb.2019.103289
ISSN/ISBN:		1879-0240 (Electronic) 0965-1748 (Linking)
Abstract:		"The Xenopus oocyte and the Human Embryonic Kidney (HEK) 293 cell expression systems are frequently used for functional characterization (deorphanization) of insect odorant receptors (ORs). However, the inherent characteristics of these heterologous systems differ in several aspects, which raises the question of whether the two systems provide comparable results, and how well the results correspond to the responses obtained from olfactory sensory neurons in vivo. Five candidate pheromone receptors were previously identified in the primitive moth Eriocrania semipurpurella (Esem) and their responses were characterized in HEK cells. We re-examined the responses of these five EsemORs in Xenopus oocytes. We showed that in both systems, EsemOR1 specifically responded to the plant volatile beta-caryophyllene. EsemOR3 responded stronger to the pheromone component (S,Z)-6-nonen-2-ol than to its enantiomer (R,Z)-6-nonen-2-ol, the second pheromone component. However, EsemOR3 also responded secondarily to the plant volatile beta-caryophyllene in the oocyte system, but not in the HEK cell system. EsemOR4 was unresponsive in the HEK cells, but responded primarily to (R,Z)-6-nonen-2-ol followed by (S,Z)-6-nonen-2-ol in the oocytes, representing a discovery of a new pheromone receptor in this species. EsemOR5 was broadly tuned in both systems, but the rank order among the most active pheromone compounds and antagonists was different. EsemOR6 showed no response to any compound in either system. We compared the results obtained in the two different heterologous systems with the activity previously recorded in vivo, and performed in situ hybridization to localize the expression of these OR genes in the antennae. In spite of similar results overall, differences in OR responses between heterologous expression systems suggest that conclusions about the function of individual ORs may differ depending on the system used for deorphanization"
Keywords:		"Animals Animals, Genetically Modified/metabolism HEK293 Cells Humans Insect Proteins/*metabolism Male Moths/*metabolism Oocytes Receptors, Odorant/*metabolism Xenopus laevis/metabolism Comparison Heterologous expression Odorant receptors Olfactory sensory;"
Notes:		"MedlineHou, Xiaoqing Zhang, Dan-Dan Yuvaraj, Jothi Kumar Corcoran, Jacob A Andersson, Martin N Lofstedt, Christer eng Research Support, Non-U.S. Gov't England 2019/11/30 Insect Biochem Mol Biol. 2020 Feb; 117:103289. doi: 10.1016/j.ibmb.2019.103289. Epub 2019 Nov 25"