Investigation of odorant receptors associated with nestmate recognition in the Argentine ant, Linepithema humile

Given the relatively poor visual acuity of compound eyes, many insects have developed alternative means for navigating their environment. For example, insects often rely on chemosensation to find food, mates, and inter- and intraspecific communication. Eusocial insects in particular have developed complex systems of pheromone communication to organize their colonies, enabling them to partition labor for foraging, brood care, and colony defense tasks to different portions of the colony. A variety of genes coding for proteins are involved in detecting these chemicals, including gustatory receptors, ionotropic receptors, and odorant receptors (ORs). Eusocial insects, and especially ants, have evolved an expanded clade of ORs in their genome, likely due to an increased reliance on pheromones compared to other insects. The ability to recognize nestmates from non-nestmates is one of the vital functions performed by these ORs, which detect hydrocarbons present on the cuticle to distinguish friend from foe. However, research into the details of nestmate recognition has been stymied due to difficulties in manipulating OR genes. Despite advances in genetic sequencing and manipulation technologies, strict reproductive divisions within most ant lineages make generating transgenic ants nearly impossible, and so we have been left with limited options to further investigate these receptors. To narrow down the ORs that could be involved in nestmate recognition in the Argentine ant (Mayr, 1868), I took a multi-pronged approach of generating tissue transcriptomes to identify ORs that are selectively upregulated in the antennae, as well as conducting a phylostratigraphic analysis to identify which OR genes arose more recently in the Argentine ant genome. While conducting these analyses, it became necessary to reannotate the set of Argentine ant OR genes, due to current published annotations not containing the full breadth of L. humile ORs. Finally, I orally administered fluorescently-labelled dsRNA to workers, and tracked the extent to which ingested dsRNA is capable of traversing the tissues of ant workers, to investigate whether RNAi is a viable method for investigating gene function for genes showing tissue-selective expression. I discovered a subset of OR genes that are highly expressed in the antennae and confirmed that dsRNA is able to reach the antennae and knock down OR gene expression through ingestion, meaning that RNA interference is a viable method for the practical study of ant OR genes and can be used to further explore how individual ORs regulate nestmate recognition.