Nanometre-range acoustic sensitivity in male and female mosquitoes

Johnston's sensory organ at the base of the antenna serves as a movement sound detector in male mosquitoes, sensing antennal vibrations induced by the flight sounds of conspecific females. Simultaneous examination of acoustically elicited antennal vibrations and neural responses in the mosquito species Toxorhynchites brevipalpis has now demonstrated the exquisite acoustic and mechanical sensitivity of Johnston's organ in males and, surprisingly, also in females. The female Johnston's organ is less sensitive than that of males. Yet it responds to antennal deflections of +/- 0.0005 degrees induced by +/- 11 nm air particle displacements in the sound field, thereby surpassing the other insect movement sound detectors in sensitivity. These findings strongly suggest that the reception of sounds plays a crucial role in the sensory ecology of both mosquito sexes.     

Active auditory mechanics in mosquitoes

In humans and other vertebrates, hearing is improved by active contractile properties of hair cells. Comparable active auditory mechanics is now demonstrated in insects. In mosquitoes, Johnston's organ transduces sound-induced vibrations of the antennal flagellum. A non-muscular 'motor' activity enhances the sensitivity and tuning of the flagellar mechanical response in physiologically intact animals. This motor is capable of driving the flagellum autonomously, amplifying sound-induced vibrations at specific frequencies and intensities. Motor-related electrical activity of Johnston's organ strongly suggests that mosquito hearing is improved by mechanoreceptor motility.     

The antennal lobe of the African malaria mosquito, Anopheles gambiae - innervation and three-dimensional reconstruction

Antibody labelling and subsequent three-dimensional reconstructions of the primary olfactory centres, the antennal lobes, of male and female African malaria mosquitoes, Anopheles gambiae, revealed 61 and 60 glomerular neuropils respectively. In addition to the small difference in number of glomeruli, sexual dimorphism was observed in both the size of the antennal lobe and of individual glomeruli. Furthermore, sexual specificity was observed within the array. Anterograde staining of afferents from peripheral olfactory organs support the reconstruction of the glomerular array. Although anterograde stainings support an organotopic organization of the antennal lobe, convergence of afferents originating from different organs into single glomeruli is observed. This finding, in both A. gambiae and A. aegypti, may shed new light upon the development and function of the olfactory system.     

Display vigour and subsequent fight performance in the siamese fighting fish,betta splendens

The display of 24 individual male Siamese fighting fish to an unresponsive stimulus conspecific was measured, and the fish were then placed together in pairs. For 11 of the 12 pairs, the outcome of the aggressive interaction which ensued was predicted by the gill cover erection durations obtained in the pre-fight isolate tests. The implications of this result for theories of display function are discussed.     

Search behavior of pheromone-stimulated potato tuber moth males (Lepidoptera: Gelechiidae)

The processes of female searching by male potato tuber moths,Phthorimaea operculella, were analyzed. The behavioral components to copulation were antennal cleaning, quiescence, walking, wing fanning, contact with female, hair brush display, copulation attempt, and copulation. Males did not always succeed in mating on their first attempt. Searching behavior of males changes to “area-restricted searching” after contact with a female. Males could, therefore, find females efficiently and copulate.     

马尾松毛虫雄蛾触角毛状感受器的细微结构

马尾松毛虫Dendrolimus punctagus(Walker)雄蛾有一对羽毛状触角。在触角鞭节的每对侧枝的内侧(迎风面)着生许多毛状感受器。每个毛状感受器由几丁质表皮毛及位于其下的三个感觉神经原和三个呈同心排列的辅助细胞-鞘原细胞、毛原细胞和膜原细胞构成。几丁质表皮毛上有许多孔。毛腔内充满感受器淋巴液。感觉神经原发出的树状突伸入毛腔,浸浴于感受器淋巴液内。这些结构特征表明它是一种司嗅觉的化学感受器。雄蛾终生不取食,推断它的嗅觉感受器主要用以感受雌蛾释放的性外激素,帮助寻找配偶。     

Interactions of pheromone with moth antennae: Adsorption,desorption and transport

Isolated antennae of the male moth Antheraea polyphemus adsorbed at least 32% of ³H-labelled pheromone molecules (E-6,Z-11 hexadecadienyl acetate) from an airstream passing the antenna. About 80% of the adsorbed molecules were caught by the long olfactory hairs (sensilla trichodea). The distal half of the hairs caught about twice as many molecules as the proximal half. About 40% of the molecules desorbed if the antennae were exposed to a clean airstream for 30 min. The adsorbed molecules were transported from the hairs towards the antennal branch. Transport due to diffusion would have a diffusion coefficient of 3 × 10^?7 cm²/s. Forty per cent of the total radioactivity per hair could be detected in receptor lymph extruded from the olfactory hairs, after an incubation time of 2 min. Dried antennae showed an increased desorption and an increased velocity of the transport along the hairs. One interpretation is that the molecules enter the receptor lymph of the intact antennae and diffuse more slowly than those on the cuticular surface. Fractional elution of fresh antennae revealed a diminishing elutability of pheromone from antennae in pentane (DEP-effect) and almost constant elutability in more polar solvents (chloroform-methanol toluene). The DEP-effect could be reversibly abolished by dehydration of the antennae. It could be shown that the DEP-effect occurs mainly on the antennal branch rather than on the hairs. Residual (uneluted) radioactivity builds up mainly on the branch.     

Investigation of the sensory organs on antennae of the horseflies Hybomitra bimaculata and Tabanus bovinus (Diptera: Tabanidae) by scanning electron microscope

Sensory organs on the antennae of the horseflies Hybomitra bimaculata Macq. and Tabanus bovinus Loew are represented by the same morphological types of sensilla. Never differences in the topographical distribution of the sensilla on antennae have been also found, which can be explained by the similarity of ecological and behavioural adaptations of these insects. First and second antennal segments are found to be supplied with tactile hairs and proprioceptors. Other antennal segments bear sensory organs of several morphological types. Short thin olfactory hairs are most numerous among them. They are present on all segments of the antennal flagellum and belong to two morphological types different by the hair length. In the upper parts of the antennal segments from third to seventh several sensilla trichoidea are present, which probably serve as tactile and taste receptors.     

Structure of isolated sensilla and sensory neurons in vitro: Observations on developing silkmoth antennae

Summary By combined enzymatic and mechanical treatment, it was possible to dissociate the sensory epithelium of developing antennae of male Antheraea polyphemus and A. pernyi silkmoths from the stage of separation of the antennal branches up to the early stages of cuticle deposition. Large numbers of entire developing trichoid sensilla were isolated. These are characterized by a large trichogen cell with a long apical, hair-forming process and a large nucleus. A cluster of 2–3 sensory neurons, enclosed by the thecogen cell, is situated in the basal region. The dendrites run past the nucleus of the trichogen cell into the apical process from which they protrude laterally. The nuclei of the tormogen and a 4^th enveloping cell can be distinguished near the base of the prospective hair. After further dissociation, only the neuron clusters remain, still enclosed by their thecogen cell and often attached to the antennal branch nerve via their axons. It is finally possible to disrupt the thecogen cells and the axons, leaving the sensory neurons with inner dendritic segments and axon stumps. The majority of these neurons can be expected to be olfactory.     

Ultrastructure of the peg and hair sensilla on the antenna of larval Aedes aegypti (L.)

The antenna of fourth instar larvae of Aedes aegypti has one peg organ of a basiconic type innervated by four neurons. The dendrites are ensheathed to near their terminations at the peg tip by an electron-dense dendritic sheath and by a cuticular sheath. They have easy communication by diffusion with the external environment only at the tip through a peripheral ensheathing membrane and six slit-channels. One of the dendrites resembles a tubular body proximally and may be mechanoreceptive. The peg generally appears to be a contact chemoreceptor. There are three antennal hairs of a typical sensillum trichodeum type innervated at the base by one neuron each. An intricate terminal mechanism at the insertion of the dendrite in the hair is described. These are believed to be tactile hairs. There are also three antennal hairs each innervated by two neurons. The dendrite from one terminates at the base similar to that of a tactile hair, and is believed to function in a similar mechanoreceptive manner. The dendrite from the second neuron extends naked along the length of the hair lumen. It is believed to be primarily chemoreceptive, in a slow-acting general sensory function. In all the sensilla there appear to be secretions produced in the junction body regions of the dendrites, and there is evidence for accumulation of secretory materials in the dendritic tips in some of the sensilla.     

Antennal glands in Psylliodes chrysocephala, and their possible role in reproductive behaviour

Abstract. The antennal morphology of Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae), an important pest of oilseed rape ( Brassica napus ), was studied. Two types of tricellular, integumentary glands were found. The common antennal glands are distributed under all sensilla-bearing parts of male and female antennae. The male-specific antennal glands are only located under a glabrous area found on antennomeres 6–10 of male beetles. The common antennal glands are synthetically active in both pre- and post-diapause adults, but the male specific antennal glands are only active in post-diapause (reproductively active) males. During studies of mating behaviour, the antennae of the males were highly active at the beginning and end of copulation, and in response to increased female activity. The male specific antennal glands may secrete a sex pheromone, and the glabrous area on the male antennae could be the release site for such a pheromone.     

Effects of blood-feeding on olfactory sensitivity of the malaria mosquito Anopheles gambiae: Application of mixed linear models to account for repeated measurements

Olfaction plays an important role in the host-seeking behavior of the malaria mosquito Anopheles gambiae. After a complete blood meal, female mosquitoes will not engage in host-seeking behavior until oviposition has occurred. We investigated if peripheral olfactory sensitivity changed after a blood meal by recording electroantennograms (EAGs) of female mosquitoes at three time points (2 h, 48 h and 72 h) to 15 volatile kairomones of either human origin or documented to emanate from oviposition sites. The EAG-sensitivity was compared with that of females of similar age post eclosion. As is common practice in electrophysiological studies, the EAG recordings were obtained by repeated stimulation of the same antennal preparations. We introduce mixed linear modeling as an improved statistical analysis for electrophysiological data. Two hours after blood ingestion, olfactory sensitivity as quantified through EAG-recording increased significantly and selectively, i.e. for seven compounds, compared to unfed females of the same age. Such short-term electrophysiological sensitization in the olfactory system as a result of feeding has not been documented before for insects. Sensitization to six compounds persisted until 48 h or 72 h post-blood meal at one or more concentrations. Desensitization was observed at 48 and 72 h pbm in response to two and three kairomones, respectively. For several compounds, sensitization at the EAG-level corresponded with sensitization found previously in single sensillum studies on olfactory neurons in antennal sensilla trichodea of An. gambiae females. These effects are likely to reflect sensitization to oviposition cues, as eggs have matured 48–72 h pbm. Knowledge of changes in olfactory sensitivity to kairomones can be applied to increase trap catches of malaria mosquitoes that have taken a blood meal and need to locate oviposition sites.     

蚊虫嗅觉识别的神经机制

蚊虫主要依赖嗅觉系统与外界环境进行化学信息交流。蚊虫通过嗅觉感受系统寻找食物、 配偶和产卵场所, 进而做出相应的行为反应。本文综述了近年来蚊虫嗅觉系统对气味信号神经传导机制的研究进展。蚊虫的嗅觉感器主要位于触角和下颚须, 触角上的毛形感器和锥形感器感受氨水、 乳酸、 羧酸类化合物等人体和其他动物释放的微量气味物质, 下颚须上的锥形感器则感受呼出的二氧化碳以及一些其他的挥发性物质; 蚊虫嗅觉感器内部有受体神经细胞, 其上分布有嗅觉受体蛋白, 蚊虫对外界环境的化学感受就是通过气味物质与这些受体蛋白互作而得以实现; 根据对不同气味物质的反应谱差异, 嗅觉神经细胞被分为不同的功能类型; 来自嗅觉神经细胞的神经信号进一步从外周传导至中枢神经中脑触角叶内的神经小球, 在此对信息进行初步的处理, 通过评估嗅觉神经细胞的反应和触角叶内的神经小球相应被激活的区域, 不同小球被分别命名; 最后, 神经信号继续整合, 由投射神经传向前脑, 最终引发一系列昆虫行为反应。这些研究从理论上剖析了气味信号在蚊虫嗅觉系统中的神经转导通路, 对于我们深刻理解蚊虫的嗅觉系统具有重要意义, 同时也有助于进一步理解其他昆虫甚至人类的气味识别机制及进行更深层次神经科学的探索。     

Regulation of sensitivity in the peripheral chemoreceptor systems for host-seeking behaviour by a haemolymph-borne factor in Aedes aegypti

The taking of a blood meal and subsequent development of eggs by a female mosquito is reported to suppress host-seeking behaviour. This change in behaviour may be partly mediated by changes in certain chemosensitive antennal afferent neurones that influence the behaviour of female mosquitoes. Electrophysiological activity of the lactic acid-excited neurons to lactic acid—a normal host-attractant substance—is depressed following a blood meal. This reduction in lactic acid sensitivity is coincident with the reported inhibition of host-seeking behaviour. The reduction in lactic acid sensitivity is reversible; this sensitivity returns to the pre-blood-fed level following oviposition. Like the inhibition of host-seeking behaviour, the reduced lactic acid sensitivity is due to a transfusable, haemolymph-borne factor. A role for the peripheral sensory system in the control of behaviour in female mosquitoes is discussed.     

Development of lactic acid-receptor sensitivity and host-seeking behaviour in newly emerged female Aedes aegypti mosquitoes

The temporal patterns for the development of sensitivity in the lactic acid-excited neurones of the antennal grooved-peg sensilla and for the initiation of host-seeking behaviour by newly emerged virgin female Aedes aegypti mosquitoes were determined and compared. A 1:1 correlation between the presence of a high sensitivity to the host attractant, lactic acid, in the lactic acid-excited neurones and the presence of host-seeking behaviour was observed. This finding supports the notion that changes in the activity of the peripheral sensory system are sufficient, although perhaps not the only means, to control host-seeking behaviour of female mosquitoes.     

The auditory system of blood-sucking mosquito females (Diptera, Culicidae): Acoustic perception during flight simulation

Mosquitoes hear with their plumose antennae which respond to the air movement caused by sound propagation and conduct vibrations to the Johnston’s organ located at the base of each antenna. Each of the two Johnston’s organs contains several tens of thousands mechanosensory cells which detect the displacements of the flagellum and transform them into electric potentials. Hearing plays a very important role in the reproductive behavior of the male mosquitoes. At the same time, our knowledge of hearing in female mosquitoes is very limited and its functional significance is obscure. In this study we measured the auditory sensitivity of female mosquitoes and investigated how the flight conditions affect their hearing. We studied mosquitoes of three species: Anopheles messeae, Aedes excrucians, and Culex pipiens pipiens. The neuronal responses were recorded with a glass microelectrode from the antennal nerve and the deutocerebral interneurons. Stimulation was applied in two modes: (1) the main stimulus against the background of flight simulation (strong vibration with the typical wingbeat frequency of a given mosquito species) and (2) only the main stimulus without the background stimulation. During the flight simulation, females demonstrated an increased sensitivity to frequencies below 200 Hz. The mean auditory receptor threshold at 80–120 Hz was 45 dB, which was 8 dB lower than that without flight simulation. An additional zone of increased sensitivity was also found at frequencies higher than the simulated wingbeat frequency (the so-called image channel). Our analysis of frequency tuning curves measured from the receptors and auditory interneurons shows that mosquito auditory neuronal complex consists of several subsystems which have different frequency tuning parameters, and suggests the possibility of spectral analysis of sounds. Three hypotheses could be proposed on the function of hearing in female mosquitoes: (1) predator avoidance, (2) detection of moving prey, and (3) intraspecific communication. Each of the hypotheses involves the ability to analyze the sound frequency spectrum and subsequent signal recognition.     

Vitellogenin synthesis in the mosquito: the role of juvenile hormone in the development of responsiveness to ecdysone

ABSTRACT. Ecdysone stimulates the synthesis of vitellogenin in the fat body of mature female mosquitoes. Preparations from newly emerged animals, however, were found to be unresponsive to ecdysone. Responsiveness developed to a maximal level during a 36-h post-emergence period of maturation. This maturation could be accelerated with juvenile hormone application, prevented by allatectomy, and restored by corpora allata implants. It is concluded that the development of fat body responsiveness to ecdysone is dependent upon previous post-emergence exposure to juvenile hormone.     

Life cycle of Amblyospora indicola (Microspora: Amblyosporidae), a parasite of the mosquito Culex sitiens and of Apocyclops sp. Copepods

The life cycle of Amblyospora indicola, a parasite of the mosquito Culex sitiens, was revealed by field observations and laboratory infection experiments conducted in Australia. In northern Queensland, infected C. sitiens larvae were often found breeding in association with two cyclopoid copepods: Apocyclops dengizicus and an undescribed species of the same genus. The latter species was found to be an intermediate copepod host of this microsporidium whereas A. dengizicus was not. One complete cycle of the parasite extends over two mosquito generations (by transovarial transmission from females with binucleate spores to their eggs) and by horizontal transmission between mosquitoes and copepods. The latter involves horizontal transmission from mosquitoes to copepods via meiospores produced in larval fat body infections and horizontal transmission from copepods to mosquitoes via uninucleate spores produced within infected copepods. Uninucleate clavate spores were formed in Apocyclops sp. nov. copepods 7-10 days after exposure to larval meiospores and were infectious to larvae of a microsporidian-free colony of C. sitiens. The development of A. indicola within mosquito larvae exposed to infected copepods is similar to that of A. dyxenoides infecting C. annulirostris. It proceeds from stages with a single nucleus to diplokaryotic binucleate cells in oenocytes. These stages persist through pupation to adult emergence after which time a proportion of male mosquitoes and female mosquitoes may develop binucleate spores without the need for a blood meal. A proportion of both male and female larval progeny of infected females with binucleate spores develop patent fat body infections via transovarial transmission and die in the fourth larval instar.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mosquito bioacoustics: Auditory processing in <Emphasis Type="Italic">Culex pipiens pipiens</Emphasis> L. Males (Diptera,Culicidae) during flight simulation

As a part of mating behavior, male mosquitoes detect and locate females by listening to the sound of their wingbeats. Up to date, the auditory physiological properties in mosquitoes were studied in steady preparations. However, the sensory organs of a flying insect are affected by strong vibrations caused by its own flight muscles and wings. This influence can sufficiently modify the perception due to the nonlinear characteristics of the receptor cells. The aim of this study was to demonstrate the effect of flight conditions on the functioning of Johnston’s organs (hearing organs) in the male mosquitoes Culex pipiens pipiens L. To simulate the flight conditions, a small amount of air around the mosquito was oscillated at 500 Hz along the dorso-ventral axis. These air oscillations affected the pinnate antennae of the mosquito, causing vibration of the antennal flagellum which, in turn, was transmitted to and sensed by Johnston’s organ. Along with the flight simulation, the mosquito was stimulated by low-amplitude sinusoidal auditory signals of different frequencies. The responses were recorded from neurons using glass microelectrodes. The auditory responses contained two rhythms of amplitude modulation which were produced due to nonlinear processes in the receptors: the first rhythm had the difference frequency of flight simulation and the stimulus; the second rhythm corresponded to the beating of heterodyne frequencies. As compared to the steady-state conditions, an additional optimum of auditory sensitivity at 540–640 Hz (the so-called image channel) was shown to appear during flight simulation. This optimum corresponds to the second harmonic of the conspecific female sound. An improvement of sensitivity by 7 dB (2.2-fold) was also observed at the main optimum (220–320 Hz). We conclude that the mechanical influence of locomotor movements not only produces noise but can also improve the sensitivity of the sensory system or even add new properties.     

Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. V. Development of the peripheral nervous system

The imaginal antenna of the male silkmoth Antheraea polyphemus is a feather-shaped structure consisting of about 30 flagellomeres, each of which gives off two pairs of side branches. During the pupal stage (lasting for 3 weeks), the antenna develops from a leaf-shaped, flattened epidermal sac ('antennal blade') via two series of incisions which proceed from the periphery towards the prospective antennal stem. The development of the peripheral nervous system was studied by staining the neurons with an antibody against horseradish peroxidase as well as by electron microscopy. The epithelium is subdivided in segmentally arranged sensillogenic regions alternating with non-sensillogenic regions. Immediately after apolysis, clusters consisting of 5 sensory neurons each and belonging to the prospective sensilla chaetica can be localized at the periphery of the antennal blade in the sensillogenic regions. During the first day following apolysis, the primordia of ca. 70 000 olfactory sensilla arise in the sensillogenic regions. Axons from their neurons are collected in segmentally arranged nerves which run towards the CNS along the dorsal as well as the ventral epidermis and are enveloped by a glial sheath. This 'primary innervation pattern' is completed within the second day after apolysis. A first wave of incisions ('primary incisions') subdivide the antennal blade into segmental 'double branches' without disturbing the innervation pattern. Then a second wave of incisions ('secondary incisions') splits the double branches into single antennal branches. During this process, the segmental nerves and their glial sheaths are disintegrated. The axons are then redistributed into single branch nerves while their glial sheath is reconstituted, forming the 'secondary', or adult, innervation pattern. The epidermis is backed by a basal lamina which is degraded after outgrowth of the axons, but is reconstituted after formation of the single antennal branches.