Midori Sakura

Distribution of dendrites of descending neurons and its implications for the basic organization of the cockroach brain

To determine precisely the brain areas from which descending neurons (DNs) originate, we examined the distribution of somata and dendrites of DNs in the cockroach brain by retrogradely filling their axons from the cervical connective. At least 235 pairs of somata of DNs were stained, and most of these were grouped into 22 clusters. Their dendrites were distributed in most brain areas, including lateral and medial protocerebra, which are major termination areas of output neurons of the mushroom body, but not in the optic and antennal lobes, the mushroom body, the central complex, or the posteroventral part of the lateral horn. The last area is the termination area of major types of olfactory projection neurons from the antennal lobe, i.e., uni‐ and macroglomerular projection neurons, so these neurons have no direct connections with DNs. The distribution of axon terminals of ascending neurons overlaps with that of DN dendrites. We propose, based on these findings, that there are numerous parallel processing streams from cephalic sensory areas to thoracic locomotory centers, many of which are via premotor brain areas from which DNs originate. In addition, outputs from the mushroom body, central complex, and posteroventral part of the lateral horn converge on some of the premotor areas, presumably to modulate the activity of some sensorimotor pathways. We propose, based on our results and documented findings, that many parallel processing streams function in various forms of reflexive and relatively stereotyped behaviors, whereas indirect pathways govern some forms of experience‐dependent modification of behavior. J. Comp. Neurol. 458:158–174, 2003.

Classical Olfactory Conditioning in the Cockroach Periplaneta americana

Abstract We established a classical conditioning procedure for the cockroach, Periplaneta americana, by which odors were associated with reward or punishment. Cockroaches underwent differential conditioning trials in which peppermint odor was associated with sucrose solution and vanilla odor was associated with saline solution. Odor preference of cockroaches was tested by allowing them to choose between peppermint and vanilla sources. Cockroaches that had undergone one set of differential conditioning trials exhibited a significantly greater preference for peppermint odor than did untrained cockroaches. Memory formed by three sets of differential conditioning trials, with an inter-trial interval of 5 min, was retained at least 4 days after conditioning. This conditioning procedure was effective even for cockroaches that had been harnessed in plastic tubes. This study shows, for the first time in hemimetaborous insects, that both freely moving and harnessed insects are capable of forming olfactory memory by classical conditioning procedure. This procedure may be useful for future electrophysiological and pharmacological studies aimed at elucidation of neural mechanisms underlying olfactory learning and memory.

Olfactory Learning and Memory in the Cockroach Periplaneta americana

Abstract The olfactory learning capability of the male cockroach Periplaneta americana was studied using an operant discriminatory conditioning paradigm. Cockroaches were highly motivated to search for a sucrose solution by giving them a diet that contained no sugar and by limiting their water supply, and they were individually trained to associate peppermint with sucrose solution and vanilla with NaCl solution. Their odor preference was tested before and after training by allowing them to choose between peppermint and vanilla sources. The time spent visiting each odor source served as a measure of odor preference. Cockroaches exhibited an innate preference for vanilla over peppermint but the preference for the latter increased after only one training session. The olfactory memory formed by a single training session remained for at least one week but significantly decayed from 2 to 24 hr after training. Memory formed by three training sessions was extremely robust, exhibiting no significant decay from one day to four weeks after training. The preference formed was easily altered by reversal training in which vanilla was associated with sucrose solution and peppermint with saline. This study shows that cockroaches have an excellent olfactory learning capability characterized by rapid acquisition, extremely long retention and easy re-writing of memory.

De Novo synthesis of CREB in a presynaptic neuron is required for synaptic enhancement involved in memory consolidation

Interaction between the activator type of cyclic AMP response element binding protein (CREB1) and the repressor type (CREB2) results in determining the emergence of long‐lasting synaptic enhancement involved in memory consolidation. However, we still do not know whether the constitutively expressed forms of CREB are enough or the newly synthesized forms are required for the synaptic enhancement. In addition, if the newly synthesized forms are needed, we must determine the time for translation of CREB from its mRNA. We applied the methods of RNA interference and real‐time polymerase chain reaction (PCR) to CREB in the cerebral giant cells of Lymnaea. The cerebral giant cells play an important role in associative learning and employ a CREB cascade for the synaptic enhancement to neurons such as the B1 motoneurons. We injected the small interfering RNA (siRNA) of CREB1 or CREB2 into the cerebral giant cells and examined the changes in amplitude of excitatory postsynaptic potential (EPSP) recorded in the B1 motoneurons. The changes in the amounts of CREB1 and CREB2 mRNAs were also examined in the cerebral giant cells. The EPSP amplitude was suppressed 15 min after injection of CREB1 siRNA, whereas that was augmented 60 min after injection of CREB2 siRNA. In the latter case, the decrease in the amount of CREB2 mRNA was confirmed by real‐time PCR. Our results showed that the de novo synthesized forms of CREB are required within tens of minutes for the synaptic enhancement in memory consolidation.

Establishment of Social Status without Individual Discrimination in the Cricket

Agonistic behavior in crickets was investigated to understand socially adaptive behavior, which assists with the understanding of the design of an artificial autonomous system in a social organization. Agonistic behavior between male crickets is released by the perception of cuticular substances of conspecific males. The degree of aggressiveness in crickets escalates from antennal fencing to tactical contact until one male quits fighting, at which time social status is established. The question of whether crickets are able to recognize one another to establish social status in an agonist interaction is worthy of pursuit. However, it would be difficult to conduct an experiment to explore this issue while using many individuals. Hence, we examined the social organization among male crickets using a two-step approach: (i) an experiment involving a simulated cricket behavior model and (ii) an experiment involving a behavior experiment using real animals. Our results suggest that crickets establish social status without recognition of their opponents.

Aggressive behavior in the antennectomized male cricket Gryllus bimaculatus

SUMMARY Male crickets (Gryllus bimaculatus) exhibit intensively defensive aggressive behavior towards attacking males most often culminating in fighting. After the fight, the loser no longer exhibits aggressiveness in a second, separate encounter with another male; rather, the defeated male exhibits avoidance behavior. Here, we investigated the role of sensory input from the antennae in male defensive aggressive behavior. When we removed antennae from males (antennectomized males), we found that they showed little aggressiveness towards each other whereas they continued to exhibit typical fighting behavior towards an intact male. In addition, in a second encounter, antennectomized losers showed significantly higher aggressiveness towards another male than did intact losers. We further found that antennectomized crickets do not utilize visual or palpal sensory input to elicit aggressive behavior. In contrast, intact males showed aspects of aggressive behavior to male cuticular substances before and after winning a fight, and if they lost a fight they showed avoidance behavior. It thus appears that antennal sensory information is crucial in the mediation of aggressive and avoidance behaviors. However, sensory inputs from the antennae are not necessary to elicit defensive aggressive behavior but are necessary to discriminate conspecific males and initiate attacks against them.

Chemical Discrimination and Aggressiveness via Cuticular Hydrocarbons in a Supercolony-Forming Ant, Formica yessensis

Background Territorial boundaries between conspecific social insect colonies are maintained through nestmate recognition systems. However, in supercolony-forming ants, which have developed an extraordinary social organization style known as unicoloniality, a single supercolony extends across large geographic distance. The underlying mechanism is considered to involve less frequent occurrence of intraspecific aggressive behaviors, while maintaining interspecific competition. Thus, we examined whether the supercolony-forming species, Formica yessensis has a nestmate recognition system similar to that of the multicolonial species, Camponotus japonicus with respect to the cuticular hydrocarbon-sensitive sensillum (CHC sensillum), which responds only to non-nestmate CHCs. We further investigated whether the sensory system reflects on the apparent reduced aggression between non-nestmates typical to unicolonial species. Methodology/Principal Findings F. yessensis constructs supercolonies comprising numerous nests and constitutes the largest supercolonies in Japan. We compared the within-colony or between-colonies’ (1) similarity in CHC profiles, the nestmate recognition cues, (2) levels of the CHC sensillar response, (3) levels of aggression between workers, as correlated with geographic distances between nests, and (4) their genetic relatedness. Workers from nests within the supercolony revealed a greater similarity of CHC profiles compared to workers from colonies outside it. Total response of the active CHC sensilla stimulated with conspecific alien CHCs did not increase as much as in case of C. japonicus, suggesting that discrimination of conspecific workers at the peripheral system is limited. It was particularly limited among workers within a supercolony, but was fully expressed for allospecific workers. Conclusions/Significance We demonstrate that chemical discrimination between nestmates and non-nestmates in F. yessensis was not clear cut, probably because this species has only subtle intraspecific differences in the CHC pattern that typify within a supercolony. Such an incomplete chemical discrimination via the CHC sensilla is thus an important factor contributing to decreased occurrence of intraspecific aggressive behavior especially within a supercolony.

Antennal RNA-sequencing analysis reveals evolutionary aspects of chemosensory proteins in the carpenter ant, Camponotus japonicus

Chemical communication is essential for the coordination of complex organisation in ant societies. Recent comparative genomic approaches have revealed that chemosensory genes are diversified in ant lineages, and suggest that this diversification is crucial for social organisation. However, how such diversified genes shape the peripheral chemosensory systems remains unknown. In this study, we annotated and analysed the gene expression profiles of chemosensory proteins (CSPs), which transport lipophilic compounds toward chemosensory receptors in the carpenter ant, Camponotus japonicus. Transcriptome analysis revealed 12 CSP genes and phylogenetic analysis showed that 3 of these are lineage-specifically expanded in the clade of ants. RNA sequencing and real-time quantitative polymerase chain reaction revealed that, among the ant specific CSP genes, two of them (CjapCSP12 and CjapCSP13) were specifically expressed in the chemosensory organs and differentially expressed amongst ant castes. Furthermore, CjapCSP12 and CjapCSP13 had a ratio of divergence at non-synonymous and synonymous sites (dN/dS) greater than 1, and they were co-expressed with CjapCSP1, which is known to bind cuticular hydrocarbons. Our results suggested that CjapCSP12 and CjapCSP13 were functionally differentiated for ant-specific chemosensory events, and that CjapCSP1, CjapCSP12, and CjapCSP13 work cooperatively in the antennal chemosensilla of worker ants.

Aggressive behavior of the white-eye mutant crickets, Gryllus bimaculatus

Aggressive behavior of white-eye mutant crickets was investigated and compared with that of wild-type crickets. In the dark, wild-type pairs performed long-lasting fights with significantly higher aggressive levels compared to those in the light. In contrast, fights between two white-eye mutants were not significantly different with those between two wild-type crickets both in duration and the aggressive levels. Ethograms of aggressive behavior showed that the mutants could show typical sequentially escalating fight with the same behavioral categories as the wild-type crickets. These results indicate that the white-eye mutants are able to express normal aggressive behavior.

Model of a sensory-behavioral relation mechanism for aggressive behavior in crickets

In order to gain better understandings how animals establish an internal mechanism, we have focused on the aggressive behavior in the male crickets, in which some different modalities of sensory inputs modify the behavioral motivation. In this research, we examined how aggressiveness changes if crickets cannot perceive normal visual inputs or antennal inputs. First, behavior of the crickets is segmented based on behavioral observation. Second, sensory input-behavior output models are selected by utilizing Minimal Description Length (MDL) criterion. Third, multi-modality sensory-behavioral experiments are realized with handicapped crickets. Last, relations between multi-modality and behavior are discussed based on the obtained parameters in the model derived with experimental results.