Ken-ichi Harano

Depression of brain dopamine and its metabolite after mating in European honeybee (Apis mellifera] queens

To explore neuro-endocrinal changes in the brain of European honeybee (Apis mellifera) queens before and after mating, we measured the amount of several biogenic amines, including dopamine and its metabolite in the brain of 6- and 12-day-old virgins and 12-day-old mated queens. Twelve-day-old mated queens showed significantly lower amounts of dopamine and its metabolite (N-acetyldopamine) than both 6- and 12-day-old virgin queens, whereas significant differences in the amounts of these amines were not detected between 6- and 12-day-old virgin queens. These results are explained by down-regulation of both synthesis and secretion of brain dopamine after mating. It is speculated that higher amounts of brain dopamine in virgin queens might be involved in activation of ovarian follicles arrested in previtellogenic stages, as well as regulation of their characteristic behaviors.

Dopamine enhances locomotor activity for mating in male honeybees (Apis mellifera L.)

Dopamine plays multiple roles in the regulation of reproduction in female honeybees where it appears to act independently of juvenile hormone (JH). In males the role of dopamine and its relationship to JH control have not been elucidated. In the present study we determined hemolymph levels of dopamine and its metabolite (N-acetyldopamine) in males at post-emergence days 0-16. The development of locomotor and flight activities were recorded over the same period. Hemolymph levels of dopamine and N-acetyldopamine were found to increase at the time of onset of mating flight activity and those of dopamine decreased thereafter. Both locomotor and flight activities increased in parallel with hemolymph dopamine levels but the increased activity levels were maintained following decline of dopamine levels. Brain and meso-metathoracic ganglia levels of dopamine showed a similar developmental profile to hemolymph dopamine levels. Locomotor activities were temporarily inhibited by injection of a dopamine-receptor antagonist (cis(Z)-flupenthixol) into the thorax, and were enhanced by injection of a dopamine-receptor agonist (6,7-ADTN). These results suggest that dopamine regulates locomotor activities for mating and plays a role downstream of JH in premature males in honeybees.

Tactile stimuli perceived by the antennae cause the isolated females to produce gregarious offspring in the desert locust, Schistocerca gregaria

Maternal determination of progeny body size and coloration in the desert locust, Schistocerca gregaria, depends on the crowding conditions experienced during the short sensitive period that occurs two to six days before the deposition of the egg pod. Solitarious (isolated-reared) females produce relatively small eggs that yield solitarious green hatchlings but, females that are exposed to crowded conditions during the sensitive period, produce larger eggs that yield the dark-colored hatchlings characteristic of gregarious forms. The present study aimed to determine the stimuli influencing the maternal determination of progeny characteristics as well as the site at which such stimuli are perceived. By exposing isolated female adults to various combinations of visual, olfactory and tactile stimuli from a crowd of other adults, we found that no crowding effects could be elicited without tactile stimulation. Coating of various body surfaces with nail polish followed by exposure to crowding stimulation suggested that female adults perceive crowding stimuli with their antennae. This finding was supported by another experiment in which the antennae were either removed or covered with wax before the isolated females were exposed to crowded conditions. Neither serotonin nor an antagonist of its receptor affected the density-dependent maternal determination of progeny characteristics when injected into isolated or crowded female adults.

Influence of age and juvenile hormone on brain dopamine level in male honeybee (Apis mellifera): Association with reproductive maturation

Dopamine (DA) is a major functional biogenic amine in insects and has been suggested to regulate reproduction in female honeybees. However, its function has not been investigated in male drones. To clarify developmental changes of DA in drones, brain DA levels were investigated at various ages and showed a similar pattern to the previously reported juvenile hormone (JH) hemolymph titer. The DA level was lowest at emergence and peaked at day 7 or 8, followed by decline. Application of JH analog increased brain DA levels in young drones (2-4-days-old), suggesting regulation of DA by JH in drones. In young drones, maturation of male reproductive organs closely matched the increase in brain DA. The dry weight of testes decreased and that of seminal vesicles increased from emergence to day 8. The dry weight of mucus glands increased up to day 4. Consequently, DA regulated by JH might have reproductive behavior and/or physiological functions in drones.

Re-examination of the roles of environmental factors in the control of body-color polyphenism in solitarious nymphs of the desert locust Schistocerca gregaria with special reference to substrate color and humidity.

This study re-examines the effects of environmental factors including substrate color, humidity, food quality, light intensity and temperature on the green-brown polyphenism, black patterning and background body color of solitarious (isolated-reared) nymphs of Schistocerca gregaria. All individuals reared in yellow-green or yellow containers became green morphs, whereas those reared in white, ivory-colored, blue, grey, brown, zinc-colored and black containers produced brown morphs in similar proportions. The intensity of black patterns was negatively correlated with the brightness of the substrate color of the containers. Humidity, which previous studies claimed controls green-brown polyphenism in this species, exerted no significant influence on either the green-brown polyphenism or the black patterning. Food quality also had little effect on body color. High temperature tended to inhibit darkening. The background body color on the thorax was greatly influenced by the substrate color of rearing containers and a close correlation was found between these two variables, indicating that, in contrast to what has been suggested by others, this species exhibits homochromy to match the body color to the substrate color of its habitat. Similar responses were observed in another strain, although some quantitative differences occurred between the two strains examined. Based on these results, a new model explaining the control of body-color polyphenism in this locust is proposed and the ecological significance of black patterns in solitarious nymphs is discussed.

A two‐step mechanism controls the timing of behaviour leading to emergence from soil in adult males of the scarab beetle Dasylepida ishigakiensis

Adults of the white grub beetle Dasylepida ishigakiensis Niijima et Kinoshita (Coleoptera: Scarabaeidae) emerge from the soil around dusk for mating on subtropical islands. The present study examines the factors controlling the emergence of males in the laboratory. There are two steps involved. Standby behaviour (i.e. insect head appears at the soil surface) can be observed for several hours before the beetles actually emerge for mating. The standby behaviour is facilitated by warm conditions, although the proportion of standby individuals is influenced not only by the temperature on that day, but also by that on the previous day. Experiments in which beetles are exposed to photoperiod and thermoperiod combinations, in and out of phase, show that temperature is more important in inducing standby and emerging behaviour than light alone. For the second step, factors such as temperature, light and the presence of the female sex pheromone determine whether males will leave the standby position and emerge onto the ground. The female sex pheromone stimulates standby beetles to exhibit emerging and wing vibration behaviours, although the effect depends on when it is presented to beetles. Beetles burrow back into the soil; this behaviour is influenced by illumination and time of the day but not by temperature. The results suggest that D. ishigakiensis possesses a sophisticated mechanism controlling male emergence from the soil.

Caste differences in dopamine-related substances and dopamine supply in the brains of honeybees (Apis mellifera L.).

We determined the mechanisms underlying caste differences in the brain levels of dopamine-related substances in adult honeybees (Apis mellifera L.). Brain levels of dopamine, DOPA (a dopamine precursor), and N-acetyldopamine (a dopamine metabolite, NADA) were significantly higher in three-day-old virgin queens than same-aged workers. Caste differences in dopamine and NADA levels were also found in the hemolymph. The in vitro enzymatic activities of DOPA decarboxylase (DDC) during dopamine synthesis in brains were not significantly different between castes. The DDC activity in adult queens was mainly found in the brain, but with lower levels of activity detected in the mandibular glands, salivary glands and ovaries. Oral application of DOPA to workers led to DOPA uptake in the brain and significantly higher dopamine and NADA levels in the brain, suggesting that dopamine synthesis could be controlled by the amount of DDC substrate. Royal jelly samples taken from queen cells had a >25-fold higher concentration of dopamine compared with honey samples collected from honey cells. However, oral application of the same concentration of dopamine did not significantly enhance the brain levels of dopamine and NADA. These results suggest that the higher levels of brain dopamine in queens compared with workers can be explained by the higher level of DDC substrate, rather than DDC activity in the brain and other tissues of queens as well as exogenous dopamine in the royal jelly.

Factors influencing adult emergence from soil and the vertical distribution of burrowing scarab beetles Dasylepida ishigakiensis

The present study investigates the emergence of adult white grub beetles Dasylepida ishigakiensis Niijima et Kinoshita (Coleoptera: Scrabaeidae) from soil as well as their burrowing behaviours. ‘Standby behaviour’ (i.e. adults come to the soil surface where they expose their heads) is shown in the field and, along with emergence behaviour, is entrained by LD photocycles. These 24‐h rhythms persist after transfer to continuous light conditions for 2 days. By contrast, beetles transferred from LD photocycles to continuous dark conditions fail to show standby behaviour; thus, it appears to be manifested only in the presence of illumination. Under dark conditions, beetles emerge completely from the soil directly at the time when standby behaviour is otherwise expected to occur. Emerged adults then burrow back into the soil before dawn. Virgin and mated males, as well as virgin females, which are expected to emerge from the soil for mating on later evenings, burrow to a relatively shallow depth (<2 cm), whereas mated females burrow deeper (2–10 cm). Soil properties such as moisture, grain size, topography and temperature influence the burrowing behaviour and the depths that the beetles reach.

Honeybee foragers adjust crop contents before leaving the hive

Upon leaving the hive, foragers carry a small amount of honey, which they subsequently consume to generate energy for flight. We investigated the relationship between waggle-phase duration and crop volume in foragers (both dancers and dance followers) leaving the hive. Our findings indicate that these variables were positively correlated in the two types of bee, suggesting that they were able to adjust the amount of food that they carry depending on the distance to a food source. We also found that dance followers left the hive with a larger amount of honey than dancers. We suggest two possible explanations: (1) dance followers have less information about the location of the food source than dancers, who have a better knowledge of the surrounding area; or (2) honeybees lack a precise calibration method for estimating energy needs from waggle-run duration. The effect of foraging experience was confirmed: bees decreased their honey load at departure with repeated trips to a sugar-syrup feeder. Honeybees showed a different pattern of change when the feeder provided soybean flour as a pollen substitute, possibly because honeybees use honey not only as an energy source but also as “glue” to form “balls” of pollen on their hind legs. Based on our observations that followers of sugar-syrup foragers carry a different amount of honey in their crop than followers of soybean-followers, we suggest that waggle dancers also convey information concerning food type.

Multiple mating, prolonged copulation and male substance in a scarab beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae).

Field and laboratory observations were carried out to understand the significance of prolonged copulation and the male substance transferred from males to females in the white grub beetle Dasylepida ishigakiensis Niijima & Kinoshita. Adults emerged from the soil in the evening and formed mating pairs on sugarcane plants, where copulation lasted for 1–3 h. Mating pairs (30%) observed in the field were accompanied by one or two additional males. Some females mated with one such male after ending copulation with the first male, showing multiple mating. After mating, males dropped to the ground to burrow into the soil earlier than females. Females received a large amount of colloidal secretion from males during mating. The amount and rate of transfer of this secretion from males to females were investigated in this study. Transfer of the secretion from males to females was completed within 30 min, and females that had mated with two males in the laboratory stored a larger amount of male secretion in the bursa copulatrix than those that had mated once. The results of both field and laboratory observations might indicate a possibility that prolonged copulation serves as a post-insemination female guarding behaviour to prevent mating by other conspecific males.