Fumio Yokohari

Projection neurons originating from thermo- and hygrosensory glomeruli in the antennal lobe of the cockroach.

Most insects are equipped with specialized thermo‐ and hygroreceptors to locate a permissible range of ambient temperature and distant water sources, respectively. In the cockroach, Periplaneta americana, cold, moist, and dry receptor cells in the antennae send axons to particular sets of two or three glomeruli in the dorsocentral part of the antennal lobe (primary olfactory center), designated DC1–3 glomeruli. However, it is not known how thermo‐ and hygrosensory signals from these glomeruli are represented in higher‐order centers, the protocerebrum, in any insect species. With the use of intracellular recording and staining techniques, we identified a new class of interneurons with dendrites almost exclusively in the DC1, DC2, or DC3 glomeruli and axons projecting to the protocerebrum in the cockroach. Remarkably, terminals of all these projection neurons (PNs) covered almost identical areas in the lateral protocerebrum (LP), although their termination areas outside the LP differed from neuron to neuron. The termination areas within the LP were distinct from, but close to, those of uniglomerular and macroglomerular PNs that transmitted signals concerning general odors and female sex pheromones, respectively. PNs originating from DC1, DC2, and DC3 glomeruli exhibited excitatory responses to cold, moist, and dry stimuli, respectively, probably due to excitatory synaptic input from cold, moist, and dry receptor cells, respectively, whereas their responses were often modulated by olfactory stimuli. These findings suggested that dorsocentral PNs participate in neural pathways that lead to behavioral responses to temperature or humidity changes. J. Comp. Neurol. 455:40–55, 2003.

Sex-specific antennal sensory system in the ant Camponotus japonicus: structure and distribution of sensilla on the flagellum

The antennae are a critically important component of the ant’s highly elaborated chemical communication systems. However, our understanding of the organization of the sensory systems on the antennae of ants, from peripheral receptors to central and output systems, is poorly understood. Consequently, we have used scanning electron and confocal laser microscopy to create virtually complete maps of the structure, numbers of sensory neurons, and distribution patterns of all types of external sensilla on the antennal flagellum of all types of colony members of the carpenter ant Camponotus japonicus. Based on the outer cuticular structures, the sensilla have been classified into seven types: coelocapitular, coeloconic, ampullaceal, basiconic, trichoid-I, trichoid-II, and chaetic sensilla. Retrograde staining of antennal nerves has enabled us to count the number of sensory neurons housed in the different types of sensilla: three in a coelocapitular sensillum, three in a coeloconic sensillum, one in an ampullaceal sensillum, over 130 in a basiconic sensillum, 50–60 in a trichoid-I sensillum, and 8–9 in a trichoid-II sensillum. The basiconic sensilla, which are cuticular hydrocarbon-receptive in the ant, are present in workers and unmated queens but absent in males. Coelocapitular sensilla (putatively hygro- and thermoreceptive) have been newly identified in this study. Coelocapitular, coeloconic, and ampullaceal sensilla form clusters and show biased distributions on flagellar segments of antennae in all colony members. The total numbers of sensilla per flagellum are about 9000 in unmated queens, 7500 in workers, and 6000 in males. This is the first report presenting comprehensive sensillar maps of antennae in ants.

Functional and topographic segregation of glomeruli revealed by local staining of antennal sensory neurons in the honeybee Apis mellifera

In the primary olfactory center of animals, glomeruli are the relay stations where sensory neurons expressing cognate odorant receptors converge onto interneurons. In cockroaches, moths, and honeybees, sensory afferents from sensilla on the anterodorsal surface and the posteroventral surface of the flagellum form two nerves of almost equal thicknesses. In this study, double labeling of the two nerves, or proximal/distal regions of the nerves, with fluorescent dyes was used to investigate topographic organization of sensory afferents in the honeybee. The sensory neurons of ampullaceal sensilla responsive to CO2, coelocapitular sensilla responsive to hygrosensory, and thermosensory stimuli and coeloconic sensilla of unknown function were characterized with large somata and supplied thick axons exclusively to the ventral nerve. Correspondingly, all glomeruli innervated by sensory tract (T) 4 received thick axonal processes exclusively from the ventral nerve. Almost all T1–3 glomeruli received a similar number of sensory afferents from the two nerves. In the macroglomerular complexes of the drone, termination fields of afferents from the two nerves almost completely overlapped; this differs from moths and cockroaches, which show heterogeneous terminations in the glomerular complex. In T1–3 glomeruli, sensory neurons originating from more distal flagellar segments tended to terminate within the inner regions of the cortical layer. These results suggest that some degree of somatotopic organization of sensory afferents exist in T1–3 glomeruli, and part of T4 glomeruli serve for processing of hygro‐ and thermosensory signals. J. Comp. Neurol. 515:161–180, 2009.

The role of antennae in removing entomopathogenic fungi from cuticle of the termite, Coptotermes formosanus.

Abstract Our previous research has shown that the termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), protects itself from entomopathogenic fungi by mutual grooming behavior. The termite removes and discards foreign organisms, such as fungal conidia, from the body surface of its nestmates by mutual grooming behavior. The role of the antennae in detecting the condia was examind here. Three entomopathogenic fungi were used, Beauveria brongniartii 782 (Saccardo) (Hypocreales), Paecilomyces fumosoroseus K3 (Wize) (Hyphomycetes), and Metarhizium anisopliae 455 Sorokin (Hyphomycetes). Termites with antennae removed conidia more efficiently than termites without antennae. There were differences between termites with and without antennae in selection of sites to be groomed on nestmates, in the length of grooming and in occurrence of grooming. Electroantennogram (EAG) responses were recorded from termite antennae and the waveforms were rather specific to the kinds of fungi used as odor sources. Termites were able to distinguish between the tested fungi in feeding tests. These results show that the antennae play important roles in the mutual grooming behavior of the termite.

Dual, multilayered somatosensory maps formed by antennal tactile and contact chemosensory afferents in an insect brain.

The antennae of most insects move actively and detect the physical and chemical composition of objects encountered by using their associated tactile sensors. Positional information is required for these sensory modalities to interpret the physical environment. Although we have a good understanding of antennal olfactory pathways, little is known about the destinations of antennal mechanosensory and contact chemosensory (gustatory) receptor neurons in the central nervous system. The cockroach Periplaneta is equipped with a pair of long, thin antennae, which are covered in bristles. The distal portions of each antenna possess about 6,500 bimodal bristles that house one tactile sensory and one to four contact chemosensory neurons. In this study, we investigated the morphologies of bimodal bristle receptor afferents by staining individual or populations of bristles. Unlike olfactory afferents, which project exclusively into the glomeruli in the ventral region of the deutocerebrum, both the presumptive mechanosensory and the contact chemosensory afferents projected into the posterior dorsal region of the deutocerebrum and the anterior region of the subesophageal ganglion. Each afferent showed multilayered segmentation and spatial occupation reflecting its three‐dimensional position in the periphery. Presumptive contact chemosensory afferents, characterized by their thin axons and unique branching pattern, occupied more medioventral positions compared with the presumptive tactile afferents. Furthermore, projection fields of presumptive contact chemosensory afferents from single sensilla tended to be segregated from each other. These observations suggest that touch and taste positional information from the antenna is precisely represented in primary centers in a modality‐specific manner. J. Comp. Neurol. 493:291–308, 2005.

Sexual Dimorphism in the Antennal Lobe of the Ant Camponotus japonicus

Abstract The carpenter ant, a social hymenopteran, has a highly elaborated antennal chemosensory system that is used for chemical communication in social life. The glomeruli in the antennal lobe are the first relay stations where sensory neurons synapse onto interneurons. The system is functionally and structurally similar to the olfactory bulbs of vertebrates. Using three-dimensional reconstruction of glomeruli and subsequent morphometric analyses, we found sexual dimorphism of the antennal lobe glomeruli in carpenter ants, Camponotus japonicus. Female workers and unmated queens had about 430 glomeruli, the highest number reported so far in ants. Males had a sexually dimorphic macroglomerulus and about 215 ordinary glomeruli. This appeared to result from a greatly reduced number of glomeruli in the postero-medial region of the antennal lobe compared with that in females. On the other hand, sexually isomorphic glomeruli were identifiable in the dorsal region of the antennal lobe. For example, large, uniquely shaped glomeruli located at the dorso-central margin of the antennal lobe were detected in all society members. The great sexual dimorphism seen in the ordinary glomeruli of the antennal lobe may reflect gender-specific tasks in chemical communications rather than different reproductive roles.

Response characteristics of two types of cold receptors on the antennae of the cockroach, Periplaneta americana L.

Summary1.There are two types of cold receptors on the antennae of Periplaneta americana. One type of cold receptor is present in a thermo- and hygroreceptive sensillum along with a moist receptor and a dry receptor, and the other is in a thermoreceptive and olfactory sensillum along with olfactory receptors. These two types of cold receptors were investigated using stimulations of constant temperature (T), stepwise decrease of temperature (ΔT), and constant rates of temperature decrease (dT/dt).2.For cold receptors in the thermo- and hygroreceptive sensilla (C-H cells), steady-state activities are higher at lower constant temperatures (T) within the range of 16–31 °C examined (n = 7). On the other hand, those of the cold receptors in the thermoreceptive and olfactory sensilla (C-O cells) have individually a broad activity maximum in the range of 18–27 °C (n = 7).3.The differential sensitivity to downward temperature steps (ΔT) of C-H cells was in the range of - 5.4 to - 12.4 (impulses/s)/°C (n = 11). It was higher at lower adapting temperature, from which changes were initiated, in the range of about 21 to 31 °C, that is, it was reflected by the steady-state activity at the initial and adapting temperature. On the other hand, the differential sensitivity of C-O cells was in the range of - 10.7 to- 15.2 (impulses/s)/°C (n = 9), and was little reflected by the steady-state activity.4.Responses to decreasing temperature in C-H cells (n = 14) are negatively related to the rate of temperature decrease (dT/dt) in the range of about 0 to - 0.16 °C/s at given temperatures, and are also affected by instantaneous temperature (T) in the range of 18 to 30 °C. This dependence is also observed in C-O cells (n = 10), but less pronounced than in C-H cells.

Further exploration into the adaptive design of the arthropod “microbrain”: I. Sensory and memory-processing systems

Abstract Arthropods have small but sophisticated brains that have enabled them to adapt their behavior to a diverse range of environments. In this review, we first discuss some of general characteristics of the arthropod “microbrain” in comparison with the mammalian “megalobrain”. Then we discuss about recent progress in the study of sensory and memory-processing systems of the arthropod “microbrain”. Results of recent studies have shown that (1) insects have excellent capability for elemental and context-dependent forms of olfactory learning, (2) mushroom bodies, higher olfactory and associative centers of arthropods, have much more elaborated internal structures than previously thought, (3) many genes involved in the formation of basic brain structures are common among arthropods and vertebrates, suggesting that common ancestors of arthropods and vertebrates already had organized head ganglia, and (4) the basic organization of sensori-motor pathways of the insect brain has features common to that of the mammalian brain. These findings provide a starting point for the study of brain mechanisms of elaborated behaviors of arthropods, many of which remain unexplored.

Exploration into the Adaptive Design of the Arthropod “Microbrain”

Abstract Arthropods have small but sophisticated brains which have enabled them to adapt their behavior to a diverse range of environments. The enormous evolutionary success of arthropods in terms of species richness and diversity depends on the sophistication of their brains. Advances in neurobiology have clarified some of the sensory and motor mechanisms of the arthropod brain, but the basic rules of computation underlying the central functions of the arthropod brain remain unknown. Consequently, it is not known how the basic design of the arthropod brain differs from, or is analogous to, that of other animals, especially mammals. In this report, we argue that characteristic features of the arthropod “microbrain” can be ascribed not only to the limited number of its constituting neurons but also to the optimization to life with a small body.

Function-specific distribution patterns of axon terminals of input neurons in the calyces of the mushroom body of the cockroach, Periplaneta americana.

Input neurons (INs) in the calyces of the mushroom bodies (MBs) of the cockroach brain were examined by single- or multiple-staining with cobalt lysine and by Golgi impregnation. Olfactory INs had axon terminals with tuft-like, button-like or spiny-blebbed arbors in specific concentric zones in calycal neuropil. INs which responded to light stimulation had thick brush-like arbors along with axonal branches extending radially along the inner layer of calycal neuropil. Some of multiglomerular INs and two types of protocerebral INs extended blebbed axonal branches to the outer surface layer of calycal neuropil or thick bush-like axonal branches with many varicosities to entire calycal neuropil. The distribution patterns of dendrites and axon terminals of INs in the calyces suggest the existence of functional subdivisions in calycal neuropil.