Circadian rhythm of locomotor activity in the Japanese honeybee, Apis cerana japonica

Abstract.  To reveal circadian characteristics and entrainment mechanisms in the Japanese honeybee Apis cerana japonica , the locomotor-activity rhythm of foragers is investigated under programmed light and temperature conditions. After entrainment to an LD 12 : 12 h photoperiodic regime, free-running rhythms are released in constant dark (DD) or light (LL) conditions with different free-running periods. Under the LD 12 : 12 h regime, activity offset occurs approximately 0.4 h after lights-off transition, assigned to circadian time (Ct) 12.4 h. The phase of activity onset, peak and offset, and activity duration depends on the photoperiodic regimes. The circadian rhythm can be entrained to a 24-h period by exposure to submultiple cycles of LD 6 : 6 h, as if the locomotive rhythm is entrained to LD 18 : 6 h. Phase shifts of delay and advance are observed when perturbing single light pulses are presented during free-running under DD conditions. Temperature compensation of the free-running period is demonstrated under DD and LL conditions. Steady-state entrainment of the locomotor rhythm is achieved with square-wave temperature cycles of 10 °C amplitude, but a 5 °C amplitude fails to entrain.     

Neurons of the rabbit midbrain reticular formation during a defensive conditioned reflex]

Analysis of unit activity of the midbrain reticular formation was carried out on alert rabbits during defensive conditioning. Most of the examined neurones exhibited phasic responses corresponding in time to the components of the evoked potential (EP) recorded in the cortical visual area in response to the "indifferent" stimulus, and to the conditioned stimulus and electric cutaneous reinforcement. The data obtained are considered from the standpoint of the Anokhin functional systems theory. A conclusion has been made regarding the participation of reticular units in providing all the basic mechanisms of the functional system of the behavioral act. Discharges of one and the same neurone may correspond to different components of the EPs to conditioned and unconditioned stimuli. In different behavioral acts a neurone may apparently participate in different systemic mechanisms.     

Octopamine modulates activity of neural networks in the honey bee antennal lobe

Neuronal plasticity allows an animal to respond to environmental changes by modulating its response to stimuli. In the honey bee (Apis mellifera), the biogenic amine octopamine plays a crucial role in appetitive odor learning, but little is known about how octopamine affects the brain. We investigated its effect in the antennal lobe, the first olfactory center in the brain, using calcium imaging to record background activity and odor responses before and after octopamine application. We show that octopamine increases background activity in olfactory output neurons, while reducing average calcium levels. Odor responses were modulated both upwards and downwards, with more odor response increases in glomeruli with negative or weak odor responses. Importantly, the octopamine effect was variable across glomeruli, odorants, odorant concentrations and animals, suggesting that the octopaminergic network is shaped by plasticity depending on an individual animal’s history and possibly other factors. Using RNA interference, we show that the octopamine receptor AmOA1 (homolog of the Drosophila OAMB receptor) is involved in the octopamine effect. We propose a network model in which octopamine receptors are plastic in their density and located on a subpopulation of inhibitory neurons in a disinhibitory pathway. This would improve odor-coding of behaviorally relevant, previously experienced odors.     

Multiplication of Kashmir bee virus in pupae of the honeybee,Apis mellifera

Laboratory investigation has shown that Kashmir bee virus (KBV) is a virulent pathogen of honeybee pupae. Inoculation with 1 × 10⁻⁸ ng of purified virus was sufficient to cause infection in some pupae, and at doses of 1 × 10⁻⁵ ng every inoculated individual became infected (ID₁₀₀). At completion of virus multiplication, between 200 and 500 μg of KBV could be recovered from each pupa. No significant relationship was found between final quantity of virus per pupa and inoculum size when the latter was greater than or equal to an ID₁₀₀. Observation by electron microscopy showed that KBV multiplied in fore- and hindgut epithelial tissue, alimentary canal musculature, epidermis and tracheal epithelium, and in hemocytes, oenocytes, and tracheal end cells. Infected cells showed a marked pathological response which included condensation of chromatin, disruption of cytoplasmic organelles, and sloughing of lobules of cytoplasm. Virus multiplication apparently took place within cytoplasmic membrane-bound vesicles. The ultrastructure of these vesicles shows little similarity to those previously described for other small RNA viruses. No evidence of KBV multiplication was found in tissues of the nervous system, even though degeneration of glial cells was observed. Electron microscopy showed that the cytoplasm of affected glial cells contained large numbers of membrane-bound vacuoles. Coalescence of these vacuoles apparently caused a separation of the nucleus and cytoplasm of cells and was followed by the progressive degeneration of each. KBV infection caused significant changes in the hemolymph osmolality of infected pupae, but no discernible changes could be seen in pupal brain morphology following induction of identical osmolality levels by inoculation with concentrated salt solutions.     

蜂群中不同亚家庭的工蜂寿命分析

在蜂群中,蜂王与多个雄蜂自然交尾,形成不同亚家庭.为了研究不同亚家庭中工蜂寿命是否有差异,我们以西方蜜蜂Apis mellifera L.为实验材料,随机从一群自然群中取270只刚孵化的工蜂,单独饲养于有蜜粉脾小蜂箱内,每天将自然死亡个体取出标记.并利用3对微卫星进行个体基因型分析,通过Matesoft软件划分亚家庭,然后分析了各亚家庭工蜂的自然寿命及生存曲线.结果表明:实验蜂群由12个亚家庭组成,其中2个亚家庭工蜂寿命与其他亚家庭存在显著差异(P＜0.05).     

Ecdysteroid biosynthesis in workers of the European honeybee Apis mellifera L

We previously reported preferential expression of genes for ecdysteroid signaling in the mushroom bodies of honeybee workers, suggesting a role of ecdysteroid signaling in regulating honeybee behaviors. The organs that produce ecdysteroids in worker honeybees, however, remain unknown. We show here that the expression of neverland and Non-molting glossy/shroud, which are involved in early steps of ecdysteroid synthesis, was enhanced in the ovary, while the expression of CYP306A1 and CYP302A1, which are involved in later steps of ecdysone synthesis, was enhanced in the brain, and the expression of CYP314A1, which is involved in converting ecdysone into active 20-hydroxyecdysone (20E), was enhanced in the brain, fat body, and ovary. In in vitro organ culture, a significant amount of ecdysteroids was detected in the culture medium of the brain, fat body, and hypopharyngeal glands. The ecdysteroids detected in the culture medium of the fat body were identified as ecdysone and 20E. These findings suggest that, in worker honeybees, cholesterol is converted into intermediate ecdysteroids in the ovary, whereas ecdysone is synthesized and secreted mainly by the brain and converted into 20E in the brain and fat body.     

Correlation between bioelectrical processes of the cortex, thalamus, midbrain reticular formation during formation of a defensive conditioned reflex in rabbits]

EEG power spectra of the sensorimotor area of the neocortex, the dorsal hippocampus, midbrain reticular formation and anteroventral thalamic nucleus, as well as corresponding coherence functions and phase spectra, undergo changes during formation and performance of defensive conditioned reflex in rabbit. The conclusion is draen that in the process of conditioning a morphofunctional system of brain structures is established including the above mentioned formations. Their functional integration occurs on the basis of theta-rhythm. The execution of a conditioned act requires isorhythmicity of electrical processes within the theta-range in the studied structures and an adequate level of their excitability, which is manifested in the dominance of 6,0 c/s frequency.     

Clique formation of super-sister honeybee workers (<Emphasis Type="Italic">Apis mellifera</Emphasis>) in experimental groups

Summary One day old honeybee workers (Apis mellifera) were observed in small experimental groups (10 workers per group). These groups were either composed of offspring workers of singly inseminated queens (super-sister groups) or multiply inseminated queens (mixed groups). The groups thus consisted of either super-sisters or a mix of super- and halfsisters. The positions of the individually labelled workers were observed with infrared sensitive video equipment over a 24 h period and analysed with digital image analysis. The spatial distribution of workers in super-sister and mixed groups differed significantly. The distance between super-sister workers was significantly less than in mixed groups (n = 339; p < 0.01). Also the distance of the workers from the group centre was significantly less in super-sister groups as compared to mixed groups (n = 3440, p<0.05). The super-sisters thus formed tighter groups than the groups including half-sisters. A genotypic analysis of the mixed groups with microsatellite DNA markers revealed that workers were significantly more frequently observed next to a super-sister rather than a half-sister (p < 0.001). Irrespective whether this results from kin recognition or not, we expect clique formation to be an important factor for the development of task specialisation among worker bees.Received 26 August 2002; revised 22 April 2003; accepted 25 July 2003.     

Detection of monohydroxy "bile" acids in the brains of guinea pigs afflicted with experimental allergic encephalomyelitis

3alpha-Hydroxy-5-cholanoic acid (lithocholic acid) and some unidentified acids (one of them perhaps 3-hydroxy-cholest-5-en-26-oic acid) have been detected in the brains of guinea pigs afflicted with experimental "allergic" encephalomyelitis. None of the acids could be identified in comparable amounts of brain tissue from normal guinea pigs. Thin-layer chromatography of the free acids, and thin-layer and gas-liquid chromatography combined with mass spectrometry of the methyl esters were used for identification. The occurrence of these acids may possibly be the result of cholesterol oxidation in the brain. The acids, or compounds related to them, may play a role in development of demyelination in experimental allergic encephalomyelitis.