Spectral sensitivities of photoreceptors and lamina monopolar cells in the dragonfly,Hemicordulia tau

Summary Five spectral types of photoreceptors with peak sensitivities at 330 nm, 410 nm, 460 nm, 525 nm and 630 nm were recorded from the ventral eye of the dragonfly, Hemicordulia tau. Often the 525 nm photoreceptors presented broader, and the 630 nm photoreceptors narrower, spectral sensitivities than would be excepted of a photopigment with the same peak sensitivity. Four types of lamina monopolar cells (cell types 1–4) were recognised from their dark-adapted spectral sensitivities and their anatomy. The anatomical identification allows tentative assignation to the monopolar cell classification from Sympetrum rubicundulum obtained using Golgi staining (Meinertzhagen and Armett-Kibel 1982). When dark-adapted, the monopolar cells had peak spectral sensitivities that were similar to single photoreceptors or appeared to pool receptor outputs, but in some cases spectral sensitivity changed markedly upon adaptation to white and to chromatic light, in one case (cell type 2) apparently switching off a UV-sensitive input.     

Tactile learning in the honeybee

Free-flying bees were conditioned on a vertical wall to a vertical tactile pattern consisting of parallel lines of grooves and elevations. The asymptote of the learning curve is reached after approximately 25 rewards. Bees can discriminate the conditioned vertical pattern from a horizontal or diagonal alternative. Angle discrimination is apparent only for relatively coarse tactile cues. The proboscis extension response of fixed bees was used to condition bees to a vertical tactile pattern which was presented to the antennae. The learning curve reaches an asymptote after 4 rewards. After 7 unrewarded extinction trials the conditioned responses are reduced to 50%. Bees show best discrimination for patterns whose edges they can scan with their antennae. The animals show a high degree of generalization by responding to an object irrespective of the trained pattern. Under laboratory conditions fixed bees can discriminate the angles and spatial wavelengths of fine tactile patterns consisting of parallel grooves. Bees can also discriminate forms and sizes of tactile patterns. They do not discriminate between different types of edges and between positive and negative forms. Accepted: 17 September 1998     

Pyrenocine A induces monopolar spindle formation and suppresses proliferation of cancer cells

Pyrenocine A, a phytotoxin, was found to exhibit cytotoxicity against cancer cells with an IC₅₀ value of 2.6–12.9 μM. Live cell imaging analysis revealed that pyrenocine A arrested HeLa cells at the M phase with characteristic ring-shaped chromosomes. Furthermore, as a result of immunofluorescence staining analysis, we found that pyrenocine A resulted in the formation of monopolar spindles in HeLa cells. Monopolar spindles are known to be induced by inhibitors of the kinesin motor protein Eg5 such as monastrol and STLC. Monastrol and STLC induce monopolar spindle formation and M phase arrest via inhibition of the ATPase activity of Eg5. Interestingly, our data revealed that pyrenocine A had no effect on the ATPase activity of Eg5 in vitro, which suggested the compound induces a monopolar spindle by an unknown mechanism. Structure-activity relationship analysis indicates that the enone structure of pyrenocine A is likely to be important for its cytotoxicity. An alkyne-tagged analog of pyrenocine A was synthesized and suppressed proliferation of HeLa cells with an IC₅₀ value of 2.3 μM. We concluded that pyrenocine A induced monopolar spindle formation by a novel mechanism other than direct inhibition of Eg5 motor activity, and the activity of pyrenocine A may suggest a new anticancer mechanism.     

The number and arrangement of elements in the lamina cartridge of the dragonfly Sympetrum rubicundulum

Summary Five monopolar cells and two long visual fibres are a consistent component of the lamina cartridge of the ventral half of the eye of the dragonfly Sympetrum rubicundulum. They communicate with the chiasm via a cartridge axon bundle comprising a minimum of ten fibres. The arrangement of these elements is documented with respect to the ommatidial photoreceptor axon bundle innervating them. These relationships are described both within the lamina cortex and in the cross-section of the underlying cartridge.     

分离自我国蜜蜂的一株新的致病螺原体及其基本特性

【目的】通过分析分离自我国患病蜜蜂体内的螺原体MF1006的基本生物学特征,初步确定其分类地位及致病性。【方法】应用暗视野显微镜和透射电子显微镜观察螺原体形态,运用常规螺原体分离和培养方法、分子生物学方法和血清学方法研究螺原体分离菌株可能的分类地位,并采用饲喂接种法研究其致病性。【结果】菌株MF1006具有典型的螺旋形和运动性,能通过0.22μm孔径的滤膜,与我国之前发现的引起蜜蜂螺原体病的参比菌株Spiroplasma melliferum CH-1的基本生物学特征差异较大。S.melliferum CH-1抗血清对其没有抑制作用。根据16S rDNA、ITS序列构建系统发育树显示,菌株MF1006与在法国发现的引起蜜蜂"五月病"的Spiroplasma apis的亲缘关系最近。此外,菌株MF1006对供试意蜂有较强的致病性。【结论】分离菌株MF1006是在我国蜜蜂体内发现的除S.melliferum以外的另一种致病螺原体。     

蜜蜂肠道菌群定殖研究进展

肠道菌群在其宿主健康中发挥着各种各样的重要功能。蜜蜂是高度社会化的昆虫,其肠道菌群与大多数昆虫明显不同,由兼性厌氧和微好氧的细菌组成,具有高度保守性和专门的核心肠道微生物群。近年来的研究表明,蜜蜂肠道微生物群在代谢、免疫功能、生长发育以及保护机体免受病原体侵袭等方面起着重要作用,并已证实肠道微生物在蜜蜂健康和疾病中起着重要作用,肠道微生物群的破坏对蜜蜂健康会产生不利影响。本文综述了蜜蜂肠道菌群的特征及菌群定殖研究进展,介绍了蜜蜂的日龄、群体、季节等对蜜蜂肠道菌群定殖的影响,探讨了宿主的功能和新陈代谢对肠道菌群的影响。     

Patterns of chromatic information processing in the lobula of the honeybee, Apis mellifera L

The honeybee, Apis mellifera L., is one of the living creatures that has its colour vision proven through behavioural tests. Previous studies of honeybee colour vision has emphasized the relationship between the spectral sensitivities of photoreceptors and colour discrimination behaviour. The current understanding of the neural mechanisms of bee colour vision is, however, rather limited. The present study surveyed the patterns of chromatic information processing of visual neurons in the lobula of the honeybee, using intracellular recording stimulated by three light-emitting diodes, whose emission spectra approximately match the spectral sensitivity peaks of the honeybee. The recorded visual neurons can be divided into two groups: non-colour opponent cells and colour opponent cells. The non-colour opponent cells comprise six types of broad-band neurons and four response types of narrow-band neurons. The former might detect brightness of the environment or function as chromatic input channels, and the latter might supply specific chromatic input. Amongst the colour opponent cells, the principal neural mechanism of colour vision, eight response types were recorded. The receptive fields of these neurons were not centre surround as observed in primates. Some recorded neurons with tonic post-stimulus responses were observed, however, suggesting temporal defined spectral opponency may be part of the colour-coding mechanisms.     

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.     

Metabolic rate during foraging in the honeybee

Summary The metabolic rate of free-flying honeybees, Apis mellifera ligustica, was determined by means of a novel respirometric device that allowed measurement of CO₂ produced by bees foraging under controlled reward at an artificial food source. Metabolic rate increased with reward (sugar flow rate) at the food source. In addition, there was no clear-cut dependence of metabolic rate on load carried during the visit, neither as crop load nor as supplementary weights attached to the thorax. The hypothesis that metabolic rate, as well as foraging and recruiting activities, depend on the motivational state of the foraging bee determined by the reward at the food source is discussed.Abbreviations CL crop load (fuel load at the FSS) - FC (=CL-W_c), fuel consumed during the visit - FSS food source simulator - FSS +dome, respirometric chamber - NVT non-visit time - TT titration time - VT visit time - W_c (=W_f-W_i) load carried at the end of the visit - W_f final weight of the forager - W_i initial weight of the forager     

Time course of color induction in the honeybee

Color induction in the honeybee is investigated in color discrimination experiments. An individual bee walks in a dark arena and is trained to a self-luminant stimulus presented from below. In the dual-choice tests the dark background is replaced by a colored induction stimulus. Choice behavior is recorded by TV camera and analyzed by computer. Successive color induction is separated from simultaneous induction by analysis of the walking paths. Only successive color induction occurs. Simultaneous effects are not observed. That is a stimulus acts as a color inducing stimulus only when the bee crosses this stimulus. Thus, the color perceived by a given eye region is found to be dependent on the viewing history, but not on the stimuli presented simultaneously on neighboring parts of the retina. Color induction in the honeybee described in terms of selective sensitivity decrease (adaptation) does not explain all behavioral effects induced by the stimulus. The time course of successive color induction is calculated from the exposure times to the induction stimulus and from the choice behavior. The data suggest that color induction is complete after a few seconds. Photoreceptor adaptation is sufficient to explain the observed time course.     

Flight of the honeybee

Summary Drag forces and lift forces acting on honeybee trunks were measured by using specially built sensitive mechanical balances. Measurements were made on prepared bodies in good and in bad flight position, with and without legs, at velocities between 0.5 and 5m·s^-1 (Reynolds numbers between 4·10² and 4·10³) and at angles of attack between-20° and +20°. From the forces drag coefficients and lift coefficients were calculated. The drag coefficient measured with a zero angle of attack was 0.45 at 3v5m·s^-1, 0.6 at 2m·s^-1, 0.9 at 1m·s^-1 and 1.35 at 0.5m·s^-1, thus demonstrating a pronounced effect of Reynolds number on drag. These values are about 2 times lower (better) than those of a drag disc with the same diameter and attacked at the same velocity. The drag coefficient (related to constant minimal frontal area) was minimal at zero angle of attack, rising symmetrically to larger (+) and smaller (-) angles of attack in a non-linear fashion. The absolute value is higher and the rise is steeper at lower speeds or Reynolds numbers, but the incremental factors are independent of Reynolds number. For example, the drag coefficient is 1.44±0.05 times higher at an angle of attack of 20° than at one of 0°. On a double-logarithmic scale the slope of the drag versus Reynolds number plot was 1.5: with decreasing Reynolds number the relationship between drag and velocity changes from quadratic (Newton's law) to linear (viscous flow). Trunk drag was not systematically increased by the legs at any velocity or Reynolds number or any angle of attack. The legs appear to shape the trunk aerodynamically, to form a relatively low-drag trunk-leg system. The body is able to generate dynamic lift. Highly significant positive linear correlations between lift coefficient and angle of attack were determined for the trunk-leg system in the typical flight position. Lift coefficient was +0.05 at zero angle of attack (possibly attained during very fast flight), +0.1 at 5° (attained during fast flight), +0.25 at +20° (attained during slow flight) and +0.55 at 45° (attained whilst changing over to hovering). Average slope c_L was 0.66±0.07, and average profile efficiency was 0.10. Non-wing lift contribution due to body form and banking only accounts for a few percent of body weight during fast flight. A non-wing lift contribution due to the legs has been demonstrated. The legs increase trunk lift by 23–24%. Reynolds number lift effects are present but of no biological significance. Force and power calculations do not support maximum flight speeds substantially higher than approximately 7m · s^-1 relative to the ambient air. At this speed body drag attains 35% and body lift 8.4% of the body weight, and parasite power is 5% of the maximum metabolic power.Abbreviations angle of attack - A area - c drag coefficient - c_L lift coefficient - D drag - F force - L lift - P power - Q quotient - Re Reynolds number - density - dliding number - O₂ oxygen consumption - W work - v kinematic viscosity - efficiency - v velocity     

Molecular characterization of hemoglobin from the honeybee Apis mellifera

Due to the prevailing importance of the tracheal system for insect respiration, hemoglobins had been considered rare exceptions in this arthropod subphylum. Here we report the identification, cloning and expression analysis of a true hemoglobin gene in the honeybee Apis mellifera (Hymenoptera). The deduced amino acid sequence covers 171 residues (19.5kDa) and harbors all globin-typical features, including the proximal and the distal histidines. The protein has no signal peptide for transmembrane transport and was predicted to localize in the cytoplasm. The honeybee hemoglobin gene shows an ancient structure, with introns in positions B12.2 and G7.0, while most other insect globins have divergent intron positions. In situ hybridization studies showed that hemoglobin expression in the honeybee is mainly associated with the tracheal system. We also observe hemoglobin expression in the Malpighi tubes and testis. We further demonstrated that hemoglobins occur in other insect orders (Hemiptera, Coleoptera, Lepidoptera), suggesting that such genes belong to the standard repertoire of an insect genome. Phylogenetic analyses show that globins evolved along with the accepted insect systematics, with a remarkable diversification within the Diptera. Although insect hemoglobins may be in fact involved in oxygen metabolism, it remains uncertain whether they carry out a myoglobin-like function in oxygen storage and delivery.     

Second-order ocellar neurons in the brain of the honeybee (Apis mellifera)

Summary Electrophoretic injection of Procion Yellow M-R4 into the ocellar tract of the worker bee has revealed the following:Two types of giant axon run from the lateral ocellus to the circumesophageal neuropile, where one branches ipsilaterally and the other contralaterally. A third type comes from the median ocellus and can be traced into the cervical connectives. The largest dendritic complex is in the circumesophageal neuropile; in addition, fiber endings have been demonstrated in the following areas: in the subretinal region, along the optic commissure, in the medulla interna, in the subesophageal ganglion and between the neurosecretory cells of the pars intercerebralis. — The giant fibers are enclosed in a glial sheath.Three types of cell body are described. One is associated with the glia; another, larger cell type comprises giant-axon somata. The third type of cell is small, and cannot yet be identified.Some of the histological results are discussed with respect to the possible function of the ocellus.     

Effect of fipronil on side-specific antennal tactile learning in the honeybee

In the honeybee, the conditioning of the proboscis extension response using tactile antennal stimulations is well suited for studying the side-specificity of learning including the possible bilateral transfer of memory traces in the brain, and the role of inhibitory networks. A tactile stimulus was presented to one antenna in association with a sucrose reward to the proboscis. The other antenna was either not stimulated (A+/0 training), stimulated with a non-reinforced tactile stimulus B (A+/B− training) or stimulated with B reinforced with sucrose to the proboscis (A+/B+ training). Memory tests performed 3 and 24 h after training showed in all situations that a tactile stimulus learnt on one side was only retrieved ipsilaterally, indicating no bilateral transfer of information. In all these groups, we investigated the effect of the phenylpyrazole insecticide fipronil by applying a sublethal dose (0.5 ng/bee) on the thorax 15 min before training. This treatment decreased acquisition success and the subsequent memory performances were lowered but the distribution of responses to the tactile stimuli between sides was not affected. These results underline the role of the inhibitory networks targeted by fipronil on tactile learning and memory processes.     

A quantitative model of successive color induction in the honeybee

Intensity discrimination experiments are performed with individual walking honeybees trained to color stimuli (UV, blue and green) of constant intensity. The choice behavior to stimuli of identical wavelength spectrum but different intensities is tested. A graded choice behavior is found. The training intensity is chosen with the highest probability in most cases. Phototaxis as well as brightness discrimination can be excluded. The choice behavior is explained exclusively by discrimination of chromaticness (hue and saturation) according to the Bezold-Brücke shift.The bees adapt to the chromatic stimuli during their choices. From the behavioral data, it is concluded that in adaptation, adjustment in photoreceptor sensitivity in one receptor also affects the sensitivity of the other receptors (co-adaptation). The linear adaptation model corresponding to the von Kries Coefficient Law used up to now to describe adaptation to white light in the honeybee does not describe this type of adaptation.A quantitative model of adaptation to chromatic stimuli extending the linear adaptation model is developed.The most reasonable mechanism of co-adaptation is optical coupling by lateral filtering. Other mechanisms such as electrical coupling are unlikely, since their effects on color vision would lead to effects inconsistent with Graßmann's Laws.     

基于TaqMan探针的蜜蜂囊状幼虫病病毒荧光PCR检测方法的建立和应用

为建立快速有效的蜜蜂囊状幼虫病检疫方法, 依据TaqMan荧光标记探针技术原理, 针对蜜蜂囊状幼虫病病毒保守序列, 设计出一对特异性引物和一条探针, 建立了一种快速检测蜜蜂囊状幼虫病病毒的荧光PCR方法。该方法对蜜蜂囊状幼虫病的检测具有较好的特异性, 与蜜蜂急性麻痹病病毒、蜜蜂慢性麻痹病病毒、蜜蜂残翼病病毒和黑蜂王台病病毒之间均无交叉反应。检测灵敏度可达1.0×10²拷贝/μL阳性质粒, 可对低病毒含量的样品进行准确检测。重复性和稳定性试验结果显示, 变异系数为1.6%, 说明该方法具有较好的重复性和稳定性。应用该方法对蜜蜂及蜂制品进行检测, 结果显示所建立的荧光PCR检测方法4 h内即可报告检测结果, 该方法具有快速、灵敏、特异及重复性好等优点, 适用于蜜蜂及其制品中蜜蜂囊状幼虫病病毒的快速检疫。     

Bipolar nanosecond electric pulses are less efficient at electropermeabilization and killing cells than monopolar pulses

Multiple studies have shown that bipolar (BP) electric pulses in the microsecond range are more effective at permeabilizing cells while maintaining similar cell survival rates as compared to monopolar (MP) pulse equivalents. In this paper, we investigated whether the same advantage existed for BP nanosecond-pulsed electric fields (nsPEF) as compared to MP nsPEF. To study permeabilization effectiveness, MP or BP pulses were delivered to single Chinese hamster ovary (CHO) cells and the response of three dyes, Calcium Green-1, propidium iodide (PI), and FM1-43, was measured by confocal microscopy. Results show that BP pulses were less effective at increasing intracellular calcium concentration or PI uptake and cause less membrane reorganization (FM1-43) than MP pulses. Twenty-four hour survival was measured in three cell lines (Jurkat, U937, CHO) and over ten times more BP pulses were required to induce death as compared to MP pulses of similar magnitude and duration. Flow cytometry analysis of CHO cells after exposure (at 15 min) revealed that to achieve positive FITC-Annexin V and PI expression, ten times more BP pulses were required than MP pulses. Overall, unlike longer pulse exposures, BP nsPEF exposures proved far less effective at both membrane permeabilization and cell killing than MP nsPEF.     

Changes in brain amine levels associated with the morphological and behavioural development of the worker honeybee

Summary Changes in biogenic amine levels associated with the morphological and behavioural development of the worker honeybee are examined.A significant increase in amine levels in the head of the honeybee is associated with transition from the larval to pupal stage. Adult emergence is also accompanied by a significant increase in 5-HT levels in the brain, but no significant change in brain dopamine (DA) levels. NADA (N-acetyldopamine) levels increase during larval and pupal development, but in contrast to both DA and 5-HT, drop significantly during the transition from pupa to adult.Levels of DA in the brain of nectar and pollen forager bees, presumed to be among the oldest adults sampled, were found to be significantly higher than in nurses, undertakers or food storers. These results suggest that an age-dependent change in amine levels occurs in the brain of the worker bee. In the optic lobes, levels of DA and 5-HT were found to be significantly higher in pollen forager bees than in all other behavioural groups. Significant differences in amine levels in the optic lobes of nectar foragers and pollen foragers indicate that some differences in amine levels occur independent of worker age. The functional significance of differences in brain amine levels and whether or not biogenic amines play a direct role in the control of honeybee behaviour has yet to be established.Abbreviations DA dopamine - 5-HT 5-hydroxytryptamine or serotonin - NADA N-acetyldopamine     

蜜蜂毒主要过敏原磷脂酶A_2研究概述

本文综述了国内外蜜蜂毒主要过敏原磷脂酶Ａ_２的研究进展,主要包括一般的蛋白质特性、作用机制、免疫学及生物学作用等。