DEET Olfactory Reception in the Yellow Fever Mosquito
Abstract
Developed almost 80 years ago, DEET is the first line of defense against the transmission of vector-borne diseases, and yet its mode of action is still a matter of controversy. Here we identified a dedicated odorant receptor that underlies DEET’s detection as a spatial repellency for the yellow fever mosquito Aedes aegypti. Using heterologous expression systems, we show that this receptor is finely tuned to DEET. CRISPR-mediated disruption impaired DEET-evoked neural activity in a defined olfactory sensillum and markedly reduced spatial avoidance at behaviorally relevant concentrations. Fluorescence-guided single-sensillum recordings revealed that DEET selectively activates a distinct olfactory sensory neuron population that projects to a single, defined glomerulus in the antennal lobe, establishing a direct receptor-to-circuit link for repellency coding. These findings uncover a dedicated sensory pathway for volatile repellent detection, revealing an unexpected specialization for a manmade chemical and providing a mechanistic framework for receptor-guided development of next-generation spatial repellents.
References
Leal, W. S. Odorant reception in insects: roles of receptors, binding proteins, and degrading enzymes. Annu. Rev. Entomol., 58, 373-391(2013).
Herre, M. et al. Non-canonical odor coding in the mosquito. Cell 185, 3104-3123.e28(2022).
Matthews,B.J., McBride,C.S., DeGennaro, M., Despo, O.& Vosshall, L.B. The neurotranscriptome of the Aedes aegypti mosquito. BMC Genomics 17, 32(2016).
Raji, J. I.& Potter, C. J. Chemosensory ionotropic receptors in human host-seeking mosquitoes. Curr. Opin. Insect. Sci. 54, 100967(2022).
Wolff, G. H.& Riffell, J. A. Olfaction, experience and neural mechanisms underlying mosquito host preference. J. Exp. Biol. 221,(2018).
Wang, Y. et al. Structural basis for odorant recognition of the insect odorant receptor OR-Orco heterocomplex. Science 384, 1453-1460(2024).
Zhao, J., Chen, A. Q., Ryu, J.& Del Marmol, J. Structural basis of odor sensing by insect heteromeric odorant receptors. Science 384, 6703(2024).
McCabe,E.T.,Barthel,W.F.,Gertler,S.I.& Hall,S.A. Insect repellents. III. N,N-diethylamides1. ACS Publications.
Debboun, M., Frances, S. P., Strickman D. Insect repellents: principles, methods, and uses.(CRC Press, Boca Raton, 2006).
Rutledge, L. C., Wirtz, R. A., Buescher, M. D.& Mehr, Z. A. Mathematical models of the effectiveness and persistence of mosquito repellents. J. Am. Mosq. Control. Assoc.1,56-62(1985).
Schofield, S.& Plourde, P. Statement on personal protective measures to prevent arthropod bites. Can. Commun. Dis. Rep.38, 1-18(2012).
Briassoulis, G., Narlioglou, M.& Hatzis, T. Toxic encephalopathy associated with use of DEET insect repellents: a case analysis of its toxicity in children. Hum. Exp. Toxicol. 20,8-14(2001).
Osimitz, T. G.& Murphy, J. V. Neurological effects associated with use of the insect repellent N, N-Diethyl-m-toluamide(DEET). J. Toxicol. Clin. 35, 435-441(1997).
Snyder, J. W., Poe, R. O., Stubbins, J. F.& Garrettson, L. K. Acute manic psychosis following the dermal application of N,N-diethyl-m-toluamide(deet) in an adult. J. Toxicol. Clin.24, 429-439(1986).
Fradin, M. S.& Day, J. F. Comparative efficacy of insect repellents against mosquito bites. New Engl. J. Med. 347, 13-18(2002).
Wylie, B. J., Hauptman, M., Woolf, A. D.& Goldman, R. H. Insect repellants during pregnancy in the era of the Zika virus. Obstetrics& Gynecology 128, 1111(2016).
Fatou, M.& Müller, P. In the arm-in-cage test, topical repellents activate mosquitoes to disengage upon contact instead of repelling them at distance. Sci. Rep. 14, 24745(2024).
Brown, M.& Hebert, A. A. Insect repellents: an overview. J. Am. Acad. Dermatol. 36, 243-249(1997).
DeGennaro, M. The mysterious multi-modal repellency of DEET. Fly 9, 45-51(2015).
Koren, G., Matsui, D.& Bailey, B. DEET-based insect repellents: safety implications for children and pregnant and lactating women. Can. Med. Assoc. J. 169,209-212(2003).
Afify,A., Betz,J.F.,Riabinina,O., Lahondre,C.& Potter,C.J. Commonly Used Insect Repellents Hide Human Odors from Anopheles Mosquitoes. Curr. Biol. 29, 3669-3680.e5(2019).
DeGennaro, M. et al. orco mutant mosquitoes lose strong preference for humans and are not repelled by volatile DEET. Nature 498, 487-491(2013).
Valbon, W. et al. Bioallethrin activates specific olfactory sensory neurons and elicits spatial repellency in Aedes aegypti. Pest Manag. Sci. 78, 438-445(2022).
Xu,P.,Choo,Y.-M., Rosa,A.D.L.& Leal, W. S. Mosquito odorant receptor for DEET and methyl jasmonate. Proc. Natl. Acad. Sci. U.S.A. 111, 16592-16597(2014).
Powell, J. R., Gloria-Soria, A.& Kotsakiozi, P. Recent history of Aedes aegypti: vector genomics and epidemiology records. BioScience 68, 854-860(2018).
Fernandez Triana, M. et al. Grapefruit-derived nootkatone potentiates GABAergic signaling and acts as a dual-action mosquito repellent and insecticide. Curr. Biol. 35, 177-186(2024).
Ghaninia, M., Ignell, R.& Hansson, B. S. Functional classification and central nervous projections of olfactory receptor neurons housed in antennal trichoid sensilla of female yellow fever mosquitoes, Aedes aegypti. Eur. J. Neurosci. 26, 1611-1623(2007).
Syed, Z.& Leal, W. S. Mosquitoes smell and avoid the insect repellent DEET. Proc. Natl. Acad. Sci. U.S.A. 105, 13598-13603(2008).
Liu,F., Chen, L., Appel, A. G.& Liu, N. Olfactory responses of the antennal trichoid sensilla to chemical repellents in the mosquito, Culex quinquefasciatus. J. Insect Physiol.59, 1169-1177(2013).
Stanczyk, N. M., Brookfield, J. F. Y., Ignell, R., Logan, J. G.& Field, L. M. Behavioral insensitivity to DEET in Aedes aegypti is a genetically determined trait residing in changes in sensillum function. Proc. Natl. Acad. Sci. U.S.A. 107, 8575-8580(2010).
Ditzen, M., Pellegrino, M.& Vosshall, L. B. Insect odorant receptors are molecular targets of the insect repellent DEET. Science 319, 1838-1842(2008).
Chahda,J. S., Soni, N., Sun, J. S., Ebrahim, S. A., Weiss, B. L.,& Carlson, J. R. The molecular and cellular basis of olfactory response to tsetse fly attractants. PLoS genetics,15(3),e1008005(2019).
Riabinina, O.,Task,D., Marr,E.et al. Organization of olfactory centres in the malaria mosquito Anopheles gambiae. Nat Commun 7, 13010(2016).
Ignell, R., Dekker, T., Ghaninia, M.& Hansson, B. S. Neuronal architecture of the mosquito deutocerebrum. J. Comp. Neurol. 493, 207-240(2005).
Shankar, S. & McMeniman, C. J. An updated antennal lobe atlas for the yellow fever mosquito Aedes aegypti. PLoS Negl. Trop. Dis. 14, e0008729 (2020).
Bohbot, J. D.& Dickens, J. C. Insect repellents: modulators of mosquito odorant receptor activity. PLoS One 5,e12138(2010).
Pellegrino, M., Steinbach, N., Stensmyr, M. C., Hansson, B. S.& Vosshall, L. B. A natural polymorphism alters odour and DEET sensitivity in an insect odorant receptor. Nature 478, 511-514(2011).
Wang, X., Chen, Z.& Liu, N. Dual mechanisms of synthetic repellents in Aedes aegypti: odorant receptor inhibition and odor volatility modulation. Entomol. Gen. 45,1475-1487(2025).
Liu,F.,Xia,X.& Liu, N. Molecular basis of N,N-diethyl-3-methylbenzamide (DEET) in repelling the common bed bug, Cimex lectularius. Front. Physiol. 8,(2017).
Goldman,O. V., DeFoe, A.E.,Qi,Y.,Jiao,Y., Weng,S.C., Wick,B.,...& Shai, N. A single-nucleus transcriptomic atlas of the adult Aedes aegypti mosquito. Cell, 188(25),7267-7290(2025).
Adavi,E.D.,Anjos, V.L.,Kotb,S., Metz,H.C.,Tian,D.,Zhao,Z.,...& McBride, C. S. Olfactory receptor coexpression and co-option in the dengue mosquito. Biorxiv.(2024).
Barbieri, F. et al. Mediterranean plants and spices as a source of bioactive essential oils for food applications: chemical characterisation and in vitro activity. Int. J. Mol. Sci. 26, 3875(2025).
Moullamri, M. et al. Salvia officinalis, Lavandula angustifolia, and Mentha pulegium essential oils: insecticidal activities and feeding deterrence against Plodia interpunctella(Lepidoptera: Pyralidae). J. Essent. Oil-Bear. Plants. 27, 16-33(2024).
Norris, E. J., Kline, J.& Bloomquist, J. R. Repellency and toxicity of vapor-active benzaldehydes against Aedes aegypti. ACS Infect. Dis. 10, 120-126(2024).
Smallergange, R. C. et al. Malaria infected mosquitoes express enhanced attraction to human odor. PLoS One 8, e63602(2013).
Mostafiz, M. M.,Jhan,P.K.,Shim,J.-K.& Lee, K.-Y. Methyl benzoate exhibits insecticidal and repellent activities against Bemisia tabaci(Gennadius)(Hemiptera: Aleyrodidae). PLOS ONE 13, e0208552(2018).
Strickland, J., Larson, N. R., Feldlaufer, M.& Zhang, A. Characterizing repellencies of methyl benzoate and its analogs against the common bed bug, Cimex lectularius. Insects 13, 1060(2022).
Singh, P. et al. Combinatorial encoding of odors in the mosquito antennal lobe. Nat Commun 14, 3539(2023).
Lahondere,C., Vinauger, C., Okubo, R. P., Wolff, G. H., Chan, J. K., Akbari, O. S.,& Riffell, J. A. The olfactory basis of orchid pollination by mosquitoes. Proc. Natl. Acad. Sci. U.S.A. 117,708-716(2020).
Raji, J. I., Konopka, J. K.& Potter, C. J. A spatial map of antennal-expressed ionotropic receptors in the malaria mosquito. Cell Rep. 42, 112101(2023).
Wang, G., Carey, A. F., Carlson, J. R. & Zwiebel, L. J. Molecular basis of odor coding in the malaria vector mosquito Anopheles gambiae. Proc. Natl. Acad. Sci. U.S.A. 107, 4418–4423 (2010).
Carey, A. F., Wang, G., Su, C.-Y., Zwiebel, L. J.& Carlson, J. R. Odorant reception in the malaria mosquito Anopheles gambiae. Nature 464, 66-71 (2010)
Dennis,E.J., Goldman, O. V.& Vosshall, L. B. Aedes aegypti mosquitoes use their legs to sense DEET on contact. Curr. Biol. 29, 1551-1556.e5(2019).
Lee,Y., Kim,S.H.& Montell, C. Avoiding DEET through insect gustatory receptors. Neuron 67, 555-561(2010).
Liu,F., Wang, Q., Xu, P., Andreazza, F., Valbon, W. R., Bandason, E., Chen, M., Yan,R.,Feng,B.,B.L.,Smith,Scott,J.G.,Takamatsu,G.,Ihara,M.,Matsuda,K., Klimavicz, J., Coats, J., Oliveira, E. E., Du, Y., Dong, K. A dual-target molecular mechanism of pyrethrum repellency against mosquitoes. Nat. Commun. 12, 2553(2021).
Chen,Z., Wang, X., Liu,F., Jia, X., Liu, N. Chemical antagonistic effects on the human odor-evoked responses of yellow fever mosquito, Aedes aegypti. Entomol. Gen.43,4(2023).
Liu,F., Liu, N. Using Single sensillum recording to detect olfactory neuron responses of bed bugs to semiochemicals. J. Vis. Exp.,107, 53337(2016).
Wang, X., Heterologous expression and functional analysis of Aedes aegypti odorant receptors to human odors in Xenopus oocytes. J. Vis. Exp.,172, e61813(2021).
Li, M., Bui, M., Yang, T., Bowman, C. S., White, B. J., Akbari, O. S. Germline Cas9 expression yields highly efficient genome engineering in a major worldwide disease vector, Aedes aegypti. Proc. Natl. Acad. Sci. U.S.A. 114, e10540-e10549(2017).
Metrics
DOI:
Submission ID:
Downloads
Posted
Versions
- 2026-05-25 (2)
- 2026-05-25 (1)
How to Cite
Download Citation
Declaration of Competing Interests
The authors declare no competing interests to disclose.
Copyright
The copyright holder for this preprint is the author/funder.
This work is licensed under a Creative Commons Attribution 4.0 International License.