Distinct antennal lobe phenotypes in the leaf-cutting ant (Atta vollenweideri).

Leaf-cutting ants (Atta vollenweideri) express a remarkable size polymorphism across the two sexual castes (queens and males) but in particular within the worker caste. Worker size is related to behavior (alloethism), separating workers into behavioral subcastes. The neuronal mechanisms underlying differences in behavior within the worker caste are still unknown. In this study, we first compared selected neuropils, in particular, the antennal lobes (AL) in males, queens, and workers. The males' ALs contain three extremely large, sex-specific glomeruli (macroglomeruli; MGs) and in total comprise fewer glomeruli (242) than the ALs of queens (about 346 glomeruli). In contrast to males, the queen ALs contain only one large glomerulus at a lateral position. The largest number of glomeruli was found in workers (396-442). In a previous paper, we described an MG in the workers' AL, and, in the second part of this study, we show that within workers two distinct, size-related AL phenotypes exist: the MG phenotype (containing a macroglomerulus) and the RG phenotype, with all glomeruli of regular size. This neuroanatomical polyphenism is established during pupal development and separates the worker caste into two neuroanatomical subcastes. Third, we investigate the functional significance of the MG in workers. By using calcium imaging to monitor activity of AL projection neurons, we show that the releaser component of the trail pheromone is represented in the same region as the MG. We propose that phenotypic trait variation in the organization of the ALs leads to differences in odor information processing that finally result in size-related differences in trail-following behavior.

Perceptual differences in trail-following leaf-cutting ants relate to body size.

Leaf-cutting ants of the genus Atta have highly size-polymorphic workers, and size is related to division of labor. We studied trail-following behavior of different-sized workers in a laboratory colony of Atta vollenweideri. For small and large workers, we measured responsiveness and preference to artificial conspecific and heterospecific pheromone trails made from poison gland extracts of A. vollenweideri and A. sexdens. Responsiveness was measured as the probability of trail-following, and preference was measured by testing the discrimination between one conspecific and one heterospecific trail. Minute amounts of the releaser component methyl-4-methylpyrrole-2-carboxylate (0.4pg/1m), present in both, conspecific and heterospecific trails, suffice to elicit trail-following behavior. Workers followed heterospecific trails, and these trails (after normalizing their concentration) were as effective as conspecific trails. Small workers were less likely to follow a trail of a given concentration than large workers. In the discrimination test, small workers preferred the conspecific trail over the heterospecific trail, whereas large workers showed no significant preference. It is suggested that large workers primarily respond to the releaser component present in both trails, whereas small workers focus more on the conspecific traits provided by the blend of components contained in the trail pheromone.

Thermal radiation as a learned orientation cue in leaf-cutting ants (Atta vollenweideri).

We explored the ability of leaf-cutting ants (Atta vollenweideri) to learn the location of a food reward by using thermal information as an orientation cue. During training of single workers, the conditioned stimulus was a distant thermal source placed frontally, 15 mm away from a platform having a leaf fragment as reward. After training, single workers were confronted with the choice between two sides, one being coupled, in a pseudo-randomized design, with a thermal stimulus heated 5 degrees C above environmental temperature. After 10 learning trials, workers significantly chose the side with the thermal stimulus. This showed that workers can use thermal information for spatial orientation in the context of foraging, which may help them to locate, for instance, highly attractive sun-exposed leaves. Thermal radiation alone as orientation cue was sufficient to allow learning, since preclusion of thermal convection during training and test did not impair workers' response. Shielding of both thorax and gaster from the thermal source did not weaken learning, suggesting the sole participation of head and antennae in temperature reception. A thermal stimulus heated 1 degrees C above environmental temperature could not be used as a learned orientation cue, even when foragers were allowed to directly contact the thermal source.

Physiology and glomerular projections of olfactory receptor neurons on the antenna of female Heliothis virescens (Lepidoptera: Noctuidae) responsive to behaviorally relevant odors.

The neurophysiology and antennal lobe projections of olfactory receptor neurons housed within short trichoid sensilla of female Heliothis virescens F. (Noctuidae: Lepidoptera) were investigated using a combination of cut-sensillum recording and cobalt-lysine staining techniques. Behaviorally relevant odorants, including intra- and inter-sexual pheromonal compounds, plant and floral volatiles were selected for testing sensillar responses. A total of 184 sensilla were categorized into 25 possible sensillar types based on odor responses and sensitivity. Sensilla exhibited both narrow (responding to few odors) and broad (responding to many odors) response spectra. Sixty-six percent of the sensilla identified were stimulated by conspecific odors; in particular, major components of the male H. virescens hairpencil pheromone (hexadecanyl acetate and octadecanyl acetate) and a minor component of the female sex pheromone, (Z)-9-tetradecenal. Following characterization of the responses, olfactory receptor neurons within individual sensilla were stained with cobalt lysine (N=39) and traced to individual glomeruli in the antennal lobe. Olfactory receptor neurons with specific responses to (Z)-9-tetradecenal, a female H. virescens sex pheromone component, projected to the female-specific central large female glomerulus (cLFG) and other glomeruli. Terminal arborizations from sensillar types containing olfactory receptor neurons sensitive to male hairpencil components and plant volatiles were also localized to distinct glomerular locations. This information provides insight into the representation of behaviorally relevant odorants in the female moth olfactory system.

A macroglomerulus in the antennal lobe of leaf-cutting ant workers and its possible functional significance.

Ants have a well-developed olfactory system, and pheromone communication is essential for regulating social life within their colonies. We compared the organization of primary olfactory centers (antennal lobes, ALs) in the brain of two closely related species of leaf-cutting ants (Atta vollenweideri, Atta sexdens). Both species express a striking size polymorphism associated with polyethism. We discovered that the ALs of large workers contain a substantially enlarged glomerulus (macroglomerulus, MG) at the entrance of the antennal nerve. This is the first description of an MG in non-sexual individuals of an insect. The location of the MG is laterally reversed in the two species, and workers of different size express a disproportional allometry of glomerular volumes. While ALs of large workers contain an MG, glomeruli in small workers are all similar in size. We further compared electroantennogram (EAG) responses to two common trail pheromone components of leaf-cutting ants: 4-methylpyrrol-2-carboxylate and 2-ethyl-3,6-dimethylpyrazine. At high concentrations the ratio of the EAG signals to 2-ethyl-3,6-dimethylpyrazine versus 4-methylpyrrol-2-carboxylate was significantly smaller in A. vollenweideri compared with the ratio of EAG signals to the same two components in A. sexdens. The differences in EAG signals and the species specific MG location in large workers provide correlative evidence that the MG may be involved in the detection of the trail pheromone.

Wind-induced ventilation of the giant nests of the leaf-cutting ant Atta vollenweideri.

To understand the significance of elaborate nest architecture for the control of nest climate, we investigated the mechanisms governing nest ventilation in a large field nest of Atta vollenweideri. Surface wind, drawing air from the central tunnels of the nest mound, was observed to be the main driving force for nest ventilation during summer. This mechanism of wind-induced ventilation has so far not been described for social insect colonies. Thermal convection, another possible force driving ventilation, contributed very little. According to their predominant airflow direction, two functionally distinct tunnel groups were identified: outflow tunnels in the upper, central region, and inflow tunnels in the lower, peripheral region of the nest mound. The function of the tunnels was independent of wind direction. Outflow of air through the central tunnels was followed by a delayed inflow through the peripheral tunnels. Leaf-cutting ants design the tunnel openings on the top of the nest with turrets which may reinforce wind-induced nest ventilation.

Prolonged reduction of salivary immunoglobulin A (sIgA) after a major academic exam.

OBJECTIVE: In a previous study we observed a continuous reduction of salivary IgA concentration ([sIgA]) during a period of academic stress. This reduction of sIgA concentration exceeded the stress period by at least 1 week. The present study aimed to replicate and extend our previous finding. In particular, we wanted to examine the time of recovery of [sIgA] alterations associated with academic stress. METHOD: Twenty-seven participants in a major medical exam and 27 controls not participating in any exam during the study provided daily saliva samples (immediately after awakening), from the 6th day prior to their last exam until the 14th day afterwards, for analysis of salivary IgA. Data were averaged for the last weeks of exams and the first and second week after exams, respectively. RESULTS: A prolonged reduction of sIgA in exam students as compared to controls was observed. Fourteen days post-stress sIgA concentrations of exam students were still significantly lower than control levels (P=0.004). No recovery was observable. At the same time exam students and controls did not differ in terms of self-reported stress and recovery. CONCLUSIONS: Psychological and immunological stress effects may be dissociated, the latter considerably exceeding the stress period. A closer look at the temporal dynamics of stress-induced immune alterations might increase our understanding of psychoimmuno relationships.

Ultrastructure and physiology of the CO2 sensitive sensillum ampullaceum in the leaf-cutting ant Atta sexdens.

The sensilla ampullacea on the apical antennomere of the leaf-cutting ant Atta sexdens were investigated regarding both their responses to CO2 and their ultrastructure. By staining the sensillum during recording, we confirmed that the sensilla ampullacea are responsible for CO2 perception. We showed that the sensory neurons of the sensilla ampullacea are continuously active without adaptation during stimulation with CO2 (test duration: 1 h). This feature should enable ants to assess the absolute CO2 concentration inside their nests. Sensilla ampullacea have been found grouped mainly on the dorso-lateral side of the distal antennal segment. Scanning and transmission electron microscopic investigations revealed that the external pore opens into a chamber which connects to the ampulla via a cuticular duct. We propose protection against evaporation as a possible function of the duct. The ampulla houses a peg which is almost as long as the ampulla and shows cuticular ridges on the external wall. The ridges are separated by furrows with cuticular pores. The peg is innervated by only one sensory neuron with a large soma. Its outer dendritic segment is enveloped by a dendritic sheath up to the middle of the peg. From the middle to the tip numerous dendritic branches (up to 100) completely fill the distal half of the peg. This is the first report of a receptor cell with highly branched dendrites and which probably is tuned to CO2 exclusively.