Cyclohexyl acetate functions like a volatile sex pheromone mimic in Caenorhabditis nematodes

Abstract Background Nematodes communicate via diverse sex pheromones, including long-range volatile signals, short-range chemical cues, and contact-dependent molecules. While the ascaroside family of small molecules that mediate short-range attraction is well characterized, the identities and roles of volatile sex pheromones (VSPs) that act over longer ranges remain unknown. Results Using GC–MS analysis of crude VSP extracts, we identified cyclohexyl acetate (CA) as a candidate mimic, sharing retention time and mass spectral features with natural VSPs. Behavioral assays demonstrated that CA acts as a concentration-dependent, male-specific attractant in Caenorhabditis. Pre-exposure to VSPs induced cross-adaptation to CA, suggesting shared sensory processing. Surprisingly, genetic and calcium imaging analyses revealed that CA perception is mediated primarily by AWCon (str-2-expressing) neurons and involves VSP chemoreceptor srd-1-independent pathways, which are distinct from the neural pathways involved in natural VSP perception. Conclusions These data indicate that CA is unlikely to be a major VSP constituent; rather, it is a structural analog that elicits male-specific attraction via a parallel sensory circuit. The endogenous source of CA in C. remanei remains unresolved; our data do not establish whether females produce CA. Its structural and behavioral mimicry provides new insights into the complexity of chemosensory signaling and the potential for interspecies chemical eavesdropping in nematode ecology.