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Functional characterization of odorant receptors from the moth Eriocrania semipurpurella : A comparison of results in the Xenopus oocyte and HEK cell systems

Hou, Xiaoqing LU ; Zhang, Dan Dan LU ; Yuvaraj, Jothi Kumar LU ; Corcoran, Jacob A. LU ; Andersson, Martin N. LU and Löfstedt, Christer LU (2020) In Insect Biochemistry and Molecular Biology 117.
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,... (More)

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 β-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 β-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.

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organization
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Contribution to journal
publication status
published
subject
keywords
Comparison, Heterologous expression, Odorant receptors, Olfactory sensory neuron, Type 0 pheromone
in
Insect Biochemistry and Molecular Biology
volume
117
article number
103289
publisher
Elsevier
external identifiers
  • scopus:85075979530
  • pmid:31778795
ISSN
0965-1748
DOI
10.1016/j.ibmb.2019.103289
language
English
LU publication?
yes
id
acdb3206-3a18-4d78-81ea-dcc9fcd1f533
date added to LUP
2019-12-19 10:37:59
date last changed
2020-03-20 03:00:20
@article{acdb3206-3a18-4d78-81ea-dcc9fcd1f533,
  abstract     = {<p>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 β-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 β-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.</p>},
  author       = {Hou, Xiaoqing and Zhang, Dan Dan and Yuvaraj, Jothi Kumar and Corcoran, Jacob A. and Andersson, Martin N. and Löfstedt, Christer},
  issn         = {0965-1748},
  language     = {eng},
  publisher    = {Elsevier},
  series       = {Insect Biochemistry and Molecular Biology},
  title        = {Functional characterization of odorant receptors from the moth Eriocrania semipurpurella : A comparison of results in the Xenopus oocyte and HEK cell systems},
  url          = {http://dx.doi.org/10.1016/j.ibmb.2019.103289},
  doi          = {10.1016/j.ibmb.2019.103289},
  volume       = {117},
  year         = {2020},
}