The spatial organization within a social insect colony is a key component of colony life. It influences individual interaction rates, resource distribution, and division of labor within the nest. Yet studies of social insect behavior are most often carried out in artificial constructions, which may change worker behavior and colony organization. We observed how workers of the ant Pheidole rhea organized brood in nests with deep chambers and textured walls that were designed to mimic their natural constructions more closely. Instead of clumping larvae into piles on the chamber floor, workers suspended fourth-instar larvae from the vertical walls and ceiling of each chamber while young larvae and pupae were clumped at the base. Fourth-instar larvae possess five rows of anchor-tipped hairs on their dorsal side, and we predicted that these hairs functioned to attach larvae to the nest walls. We gave larvae “haircuts,” where only the anchor-tipped hairs were removed, and then tested their ability to adhere to a textured surface raised to an angle of 90° and then 120° with respect to the horizontal plane. Larvae whose hairs had been clipped came unattached in almost all trials, while larvae whose hairs remained intact stayed attached. This confirmed that anchor-tipped hairs functioned to attach larvae to the walls of the nest. The presence of anchor-tipped hairs is widespread and has been documented in at least 22 genera from the ant subfamily Myrmicinae, including species that occur in a variety of environments and represent a broad range of nesting habits. Based on our results, it is likely that many species exhibit this larval hanging behavior, and this could impact colony characteristics such as spatial organization and the care of developing larvae by nurse workers.

Octopamine (OA) underlies reinforcement during appetitive conditioning in the honey bee and fruit fly, acting via different subtypes of receptors. Recently, antibodies raised against a peptide sequence of one honey bee OA receptor, AmOA1, were used to study the distribution of these receptors in the honey bee brain (Sinakevitch et al., 2011). These antibodies also recognize an isoform of the AmOA1 ortholog in the fruit fly (OAMB, mushroom body OA receptor). Here we describe in detail the distribution of AmOA1 receptors in different types of neurons in the honey bee and fruit fly antennal lobes. We integrate this information into a detailed anatomical analysis of olfactory receptor neurons (ORNs), uni- and multi-glomerular projection neurons (uPNs, and mPNs) and local interneurons (LNs) in glomeruli of the antennal lobe. These neurons were revealed by dye injection into the antennal nerve, antennal lobe, medial and lateral antenno-protocerbral tracts (m-APT and l-APT), and lateral protocerebral lobe (LPL) by use of labeled cell lines in the fruit fly or by staining with anti-GABA. We found that ORN receptor terminals and uPNs largely do not show immunostaining for AmOA1. About seventeen GABAergic mPNs leave the antennal lobe through the ml-APT and branch into the LPL. Many, but not all, mPNs show staining for AmOA1. AmOA1 receptors are also in glomeruli on GABAergic processes associated with LNs. The data suggest that in both species one important action of OA in the antennal lobe involves modulation of different types of inhibitory neurons via AmOA1 receptors. We integrated this new information into a model of circuitry within glomeruli of the antennal lobes of these species.