The arrangement of anatomically separated systems for information about general and pheromone odorants is well documented at the initial levels of the olfactory pathway both in vertebrates and insects. In the primary olfactory center of the moth brain, for example, a few enlarged glomeruli situated dorsally, at the entrance of the antennal nerve, are devoted to information about female-produced substances whereas a set of more numerous ordinary glomeruli (OG) receives input about general odorants. Heliothine moths are particularly suitable for studying central chemosensory mechanisms not only because of their anatomically separated systems for plant odors and pheromones but also due to their use of female-produced substances in communication across the species. Thus, the male-specific system of heliothine moths includes two sub-arrangements, one ensuring attraction and mating behavior by carrying information about pheromones released by conspecifics, and the other inhibition of attraction via signal information emitted from heterospecifics. Based on previous tracing experiments, a general chemotopic organization of the male-specific glomeruli has been demonstrated in a number of heliothine species. As compared to the well explored organization of the moth antennal lobe (AL), demonstrating a non-overlapping representation of the biologically relevant stimuli, less is known about the neural arrangement residing at the following synaptic level, i.e., the mushroom body calyces and the lateral horn. In the study presented here, we have labeled physiologically characterized antennal-lobe projection neurons in males of the two heliothine species, Heliothis virescens and Helicoverpa assulta, for the purpose of mapping their target regions in the protocerebrum. In order to compare the representation of plant odors, pheromones, and interspecific signals in the higher brain regions of each species, we have created standard brain atlases and registered three-dimensional models of distinct uniglomerular projection neuron types into the relevant atlas.

As in other insects, three main tracts in the moth brain form parallel connections between the antennal lobe and the protocerebrum. These tracts, which consist of the antennal-lobe projection-neuron axons, target two main areas in the protocerebrum, the calyces of the mushroom bodies and the lateral horn. In spite of the solid neuroanatomical knowledge already established, there are still unresolved issues regarding the antennal-lobe tracts of the moth. One is the proportion of lateral-tract neurons targeting the calyces. In the study presented here, we have performed both retrograde and anterograde labeling of the antennal-lobe projection neurons in the brain of the moth, Heliothis virescens. The results from the retrograde staining, obtained by applying dye in the calyces, demonstrated that the direct connection between the antennal lobe and this neuropil is maintained primarily by the medial antennal-lobe tract; only a few axons confined to the lateral tract were found to innervate the calyces. In addition, these staining experiments, which allowed us to explore the arborization pattern of labeled neurons within the antennal lobe, resulted in new findings regarding anatomical arrangement of roots and cell body clusters linked to the medial tract. The results from the anterograde staining, obtained by applying dye into the antennal lobe, visualized the total assembly of axons passing along the antennal-lobe tracts. In addition to the three classical tracts, we found a transverse antennal-lobe tract not previously described in the moth. Also, these staining experiments revealed an organized neuropil in the lateral horn formed by terminals of the four antennal-lobe tracts.