Use of Visual Information by Ant Species Occurring in Similar Urban Anthropogenic Environments.

Many insects, including ants, are known to respond visually to conspicuous objects. In this study, we compared orientation in an arena containing only a black target beacon as local information in six species of ants of widely varying degree of phylogenic relatedness, foraging strategy, and eye morphology (Aphaenogaster, Brachyponera, Camponotus, Formica, and two Lasius spp.), often found associated in similar urban anthropogenic habitats. Four species of ants displayed orientation toward the beacon, with two orienting toward it directly, while the other two approached it via convoluted paths. The two remaining species did not show any orientation with respect to the beacon. The results did not correlate with morphological parameters of the visual systems and could not be fully interpreted in terms of the species' ecology, although convoluted paths are linked to higher significance of chemical signals. Beacon aiming was shown to be an innate behavior in visually naive Formica workers, which, however, were less strongly attracted to the beacon than older foragers. Thus, despite sharing the same habitats and supposedly having similar neural circuits, even a very simple stimulus-related behavior in the absence of other information can differ widely in ants but is likely an ancestral trait retained especially in species with smaller eyes. The comparative analysis of nervous systems opens the possibility of determining general features of circuits responsible for innate and possibly learned attraction toward particular stimuli.

Odorant concentration differentiator for intermittent olfactory signals.

Animals need to discriminate differences in spatiotemporally distributed sensory signals in terms of quality as well as quantity for generating adaptive behavior. Olfactory signals characterized by odor identity and concentration are intermittently distributed in the environment. From these intervals of stimulation, animals process odorant concentration to localize partners or food sources. Although concentration-response characteristics in olfactory neurons have traditionally been investigated using single stimulus pulses, their behavior under intermittent stimulus regimens remains largely elusive. Using the silkmoth (Bombyx mori) pheromone processing system, a simple and behaviorally well-defined model for olfaction, we investigated the neuronal representation of odorant concentration upon intermittent stimulation in the naturally occurring range. To the first stimulus in a series, the responses of antennal lobe (AL) projection neurons (PNs) showed a concentration dependence as previously shown in many olfactory systems. However, PN response amplitudes dynamically changed upon exposure to intermittent stimuli of the same odorant concentration and settled to a constant, largely concentration-independent level. As a result, PN responses emphasized odorant concentration changes rather than encoding absolute concentration in pulse trains of stimuli. Olfactory receptor neurons did not contribute to this response transformation which was due to long-lasting inhibition affecting PNs in the AL. Simulations confirmed that inhibition also provides advantages when stimuli have naturalistic properties. The primary olfactory center thus functions as an odorant concentration differentiator to efficiently detect concentration changes, thereby improving odorant source orientation over a wide concentration range.

Constancy and variability of glomerular organization in the antennal lobe of the silkmoth.

We investigated the anatomical organization of glomeruli in the antennal lobes (ALs) of male silkmoths. We reconstructed 10 different ALs and established an identification procedure for individual glomeruli by using size, shape, and position relative to anatomical landmarks. Quantitative analysis of these morphological characteristics supported the validity of our identification strategy. The glomerular organization of the ALs was roughly conserved between different ALs. However, we found individual variations that were reproducibly observed. The combination of a digital atlas with other experimental techniques, such as electrophysiology, optical imaging, and genetics, should facilitate a more in-depth analysis of sensory information processing in silkmoth ALs.

Development and application of a neuroinformatics environment for neuroscience and neuroethology.

Insect brains are excellent models for analyzing neuronal function in moderately complex central nervous systems due to the vast potential they offer for revealing the intricate details of the workings of a biological neural network. For a systematic approach to understanding neuronal mechanisms, it is important to integrate research results from various fields, such as morphology, physiology and immunohistochemistry. We are developing a database system, the Bombyx Neuron Database (BoND) for assembling and sharing experimental and analytical data. The system is designed and developed based on experimental data, mostly obtained from intracellular recordings. A new WWW technology, CMS (Content Management System), was implemented in our system. That is, PHP-based CMS, XOOPS, provides several functions for web-based database management, for instance, user accounting, web page designing and data backup. The BoND was developed by our original database module of XOOPS, in order to deal with electrophysiological and anatomical data. Research resources from various fields are combined in the database for realizing a conjunction of experiments and analysis, which will assist progress in understanding neural network mechanisms as a virtual laboratory.

Morphology and physiology of antennal lobe projection neurons in the hawkmoth Agrius convolvuli.

The neuronal pathways involved in the processing of sex pheromone information were investigated in the hawkmoth Agrius convolvuli (Lepidoptera: Sphingidae), which uses (E,E)-11,13-hexadecadienal (E11,E13-16:Ald) as the single sex pheromone component. We first clarified the anatomical organization of the antennal lobe of A. convolvuli. Subsequently, central neurons in the antennal lobe that responded to E11,E13-16:Ald were identified. The dendritic processes of these neurons were confined within a specific glomerulus (cumulus) in the antennal lobe. The axons of these neurons projected to the inferior lateral protocerebrum and mushroom body calyx. Although the anatomical organization and morphology of individual neurons in A. convolvuli were similar to other species in the superfamily Bombycoidea, the use of cumulus as the single pathway for sex pheromone information processing was characteristic to this species.

Postsynaptic odorant concentration dependent inhibition controls temporal properties of spike responses of projection neurons in the moth antennal lobe.

Although odorant concentration-response characteristics of olfactory neurons have been widely investigated in a variety of animal species, the effect of odorant concentration on neural processing at circuit level is still poorly understood. Using calcium imaging in the silkmoth (Bombyx mori) pheromone processing circuit of the antennal lobe (AL), we studied the effect of odorant concentration on second-order projection neuron (PN) responses. While PN calcium responses of dendrites showed monotonic increases with odorant concentration, calcium responses of somata showed decreased responses at higher odorant concentrations due to postsynaptic inhibition. Simultaneous calcium imaging and electrophysiology revealed that calcium responses of PN somata but not dendrites reflect spiking activity. Inhibition shortened spike response duration rather than decreasing peak instantaneous spike frequency (ISF). Local interneurons (LNs) that were specifically activated at high odorant concentrations at which PN responses were suppressed are the putative source of inhibition. Our results imply the existence of an intraglomerular mechanism that preserves time resolution in olfactory processing over a wide odorant concentration range.

Concentric zones for pheromone components in the mushroom body calyx of the moth brain.

The spatial distribution of input and output neurons in the mushroom body (MB) calyx was investigated in the silkmoth Bombyx mori. In Lepidoptera, the brain has a specialized system for processing sex pheromones. How individual pheromone components are represented in the MB has not yet been elucidated. Toward this end, we first compared the distribution of the presynaptic boutons of antennal lobe projection neurons (PNs), which transfer odor information from the antennal lobe to the MB calyx. The axons of PNs that innervate pheromonal glomeruli were confined to a relatively small area within the calyx. In contrast, the axons of PNs that innervate nonpheromonal glomeruli were more widely distributed. PN axons for the minor pheromone component covered a larger area than those for the major pheromone component and partially overlapped with those innervating nonpheromonal glomeruli, suggesting the integration of the minor pheromone component with plant odors. Overall, we found that PN axons innervating pheromonal and nonpheromonal glomeruli were organized into concentric zones. We then analyzed the dendritic fields of Kenyon cells (KCs), which receive inputs from PNs. Despite the strong regional localization of axons of different PN classes, the dendrites of KCs were less well classified. Finally, we estimated the connectivity between PNs and KCs and suggest that the dendritic field may be organized to receive different amounts of pheromonal and nonpheromonal inputs. PNs for multiple pheromone components and plant odors enter the calyx in a concentric fashion, and they are read out by the elaborate dendritic field of KCs.

Development of a scheme and tools to construct a standard moth brain for neural network simulations.

Understanding the neural mechanisms for sensing environmental information and controlling behavior in natural environments is a principal aim in neuroscience. One approach towards this goal is rebuilding neural systems by simulation. Despite their relatively simple brains compared with those of mammals, insects are capable of processing various sensory signals and generating adaptive behavior. Nevertheless, our global understanding at network system level is limited by experimental constraints. Simulations are very effective for investigating neural mechanisms when integrating both experimental data and hypotheses. However, it is still very difficult to construct a computational model at the whole brain level owing to the enormous number and complexity of the neurons. We focus on a unique behavior of the silkmoth to investigate neural mechanisms of sensory processing and behavioral control. Standard brains are used to consolidate experimental results and generate new insights through integration. In this study, we constructed a silkmoth standard brain and brain image, in which we registered segmented neuropil regions and neurons. Our original software tools for segmentation of neurons from confocal images, KNEWRiTE, and the registration module for segmented data, NeuroRegister, are shown to be very effective in neuronal registration for computational neuroscience studies.

Ca2+ imaging of identifiable neurons labeled by electroporation in insect brains.

Identifiable neurons in the silkmoth brain were studied physiologically after loading of Ca indicator by local electroporation. Small groups of neurons with projections in confined regions of the antennal lobe were labeled, and Ca imaging showed differences in the dose-response characteristics between projection neurons of the macroglomerular complex that responded to different pheromone components. Compared with projection neurons, local interneurons showed shorter response latencies. Targeted labeling of neurons innervating restricted regions of neuropil by local electroporation is a powerful method that will be generally useful for elucidating details of the functional circuitry in insect brains.

Reconstruction of virtual neural circuits in an insect brain.

The reconstruction of large-scale nervous systems represents a major scientific and engineering challenge in current neuroscience research that needs to be resolved in order to understand the emergent properties of such systems. We focus on insect nervous systems because they represent a good compromise between architectural simplicity and the ability to generate a rich behavioral repertoire. In insects, several sensory maps have been reconstructed so far. We provide an overview over this work including our reconstruction of population activity in the primary olfactory network, the antennal lobe. Our reconstruction approach, that also provides functional connectivity data, will be refined and extended to allow the building of larger scale neural circuits up to entire insect brains, from sensory input to motor output.