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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.

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Dopamine has been suggested to be involved in physiological and/or behavioural changes triggered by mating in European honeybee ( Apis mellifera ) queens but its specific role remains unclear. In the present study, the amounts of dopamine (DA) and its metabolite, N -acetyldopamine (NADA) are measured, in queens of various ages to clarify the association with locomotor activity. The effects of DA receptor agonist/antagonist drugs on locomotor activity are further investigated. Brain levels of DA and NADA are relatively constant during the period before mating when locomotor activity reportedly increases with age but decreases in 1year-old laying queens with low locomotor activity. Reduced DA and NADA levels are also found in haemolymph of 1 -3-month-old laying queens. When a DA receptor agonist or antagonist is injected into 6-day-old virgin queens, locomotor activity levels increase significantly with 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (agonist), and decreased with cis( Z )-flupenthixol (antagonist). These results suggest that DA systems are involved in the motor control of honeybee queens, and that the decline in DA levels reduces locomotor activity after mating but increased locomotor activity before mating may be independent of DA levels. , Physiological Entomology, 33, 395-399 13 8 8398 K. Harano et al.

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Caste-specific differences in the dopaminergic systems of social insects assist in maintaining caste-specific behavior. To determine how caste differences in the honey bee occur during metamorphosis, a number of comparative analyses between castes were performed including comprehensive quantification of: levels of dopamine and its metabolite in the brain, the gene expression levels of enzymes involved in dopamine biosynthesis and conversion as well as expression levels of dopamine receptors and a dopamine transporter. Dopamine levels standardized to the protein contents of a whole brain at the day of eclosion in queens were 3.6-fold higher than those in workers. Dopamine levels increased until eclosion (7 days) in queens, whereas those in workers increased until 5–6 days before eclosion and then maintained until eclosion (10 days). These caste-specific dopamine dynamics in the brain were supported by the higher expression of genes (Amddc and Amth) encoding enzymes involved in dopamine synthesis in queens. The distribution of cells expressing Amddc in the brain revealed that soma clusters of dopaminergic cells were similar between castes at 7–8 days after pupation, suggesting the upregulation of Amddc expression in some cells in queens rather than addition of cell clusters. In contrast, genes encoding dopamine receptors were downregulated in queens or showed similar expression levels between castes. The expression of genes encoding dopamine transporters did not differ between castes. These results reveal the developmental process of caste-specific dopaminergic systems during metamorphosis in the honey bee, suggesting caste-specific behavior and division of reproduction in this highly eusocial species.