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Impacts of OrX and cAMP-insensitive Orco to the insect olfactory heteromer activity

Abstract

Insect odorant receptors (ORs) have been suggested to function as ligand-gated cation channels, with OrX/Orco heteromers combining ionotropic and metabotropic activity. The latter is mediated by different G proteins and results in Orco self-activation by cyclic nucleotide binding. In this contribution, we co-express the odor-specific subunits DmOr49b and DmOr59b with either wild-type Orco or an Orco-PKC mutant lacking cAMP activation heterologously in mammalian cells. We show that the characteristics of heteromers strongly depend on both the OrX type and the coreceptor variant. Thus, methyl acetate-sensitive Or59b/Orco demonstrated 25-fold faster response kinetics over o-cresol-specific Or49b/Orco, while the latter required a 10–100 times lower ligand concentration to evoke a similar electrical response. Compared to wild-type Orco, Orco-PKC decreased odorant sensitivity in both heteromers, and blocked an outward current rectification intrinsic to the Or49b/Orco pair. Our observations thus provide an insight into insect OrX/Orco functioning, highlighting their natural and artificial tuning features and laying the groundwork for their application in chemogenetics, drug screening, and repellent design.

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Data availability

The data supporting the findings of this study are available within the manuscript and its supplementary materials. The raw data are available from the corresponding author on request.

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Acknowledgements

We sincerely thank Dr. Maksim Erokhin and Dr. Dieter Wicher for their help with the DNA templates.

Funding

This work was supported by the Russian Foundation for Basic Research (RFBR) Grant 18-34-20087. Electrophysiology experiments were supported by Russian Science Foundation Grant 20-15-00408. L.A.K. was supported by the Russian Foundation for Basic Research (RFBR) Grant 19-34-90140 for PhD-students.

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AMB and ESN managed the project; AMB, ESN, PMB and KAL conceived and designed the experiments; DVK, LAK, and ELS performed the genetic engineering, cell culture and fluorescence microscopy experiments; VOI and ESN performed the electrophysiology measurements and analyzed the data; AMB, PMB, LAK acquired funding; AMB, VOI, ESN, and DVK wrote the paper. All authors analyzed the results and approved the final version of the manuscript.

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Correspondence to Alexey M. Bogdanov.

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Kolesov, D.V., Ivanova, V.O., Sokolinskaya, E.L. et al. Impacts of OrX and cAMP-insensitive Orco to the insect olfactory heteromer activity. Mol Biol Rep 48, 4549–4561 (2021). https://doi.org/10.1007/s11033-021-06480-0

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Keywords

  • Odorant receptors
  • Ionotropic transduction
  • Metabotropic transduction
  • Insect olfaction
  • Chemoreception
  • Orco