Mechanism of inhibitory action by salts in the feeding behaviour of the blowfly, Phormia regina

Blowfly proboscis extension during stimulation of labellar sugar receptors can be inhibited by the presence of salt. The possibility that the salt receptor might initiate a central nervous inhibitory state is investigated behaviourally and electrophysiologically using simultaneous recordings from labellar chemoreceptors and the extensor muscle of the haustellum. While a mixture of 100 mM sucrose and 4 M NaCl applied to a single sensillum does cause inhibition, the same compounds applied separately simultaneously on separate sensilla do not. A mixture of 100 mM sucrose and 4 M NaCl does not produce central nervous effects such as motor response decrement to repeated stimulation; nor does it produce an enhanced motor response resulting from cross-channel summation when applied simultaneously with 100 mM sucrose on another sensillum. These results argue that the inhibitory effect of mixtures containing sugar and salt can be explained by inhibition of the sugar receptor without having to invoke a central inhibitory mechanism.     

Mode of action of straight chain hydrocarbons on primary chemoreceptors of the blowfly, Phormia regina

The fundamental actions of straight-chain hydrocarbons on the three known primary chemoreceptor cell types of the blowfly, Phormia regina, were studied in quantitative terms. Lower alcohols and long-chain amines act in two stages: first a reversible inhibition, and then injury, of the salt, water, and sugar receptors in the labellar sensilla. The primary effects on salt and water receptors resemble hydrocarbon narcosis of nerve. Effects on sugar receptors, when analyzed kinetically, superficially resemble competitive inhibition. Other evidence, however, indicates that a non-specific effect on sugar receptor sites is more likely. The electrophysiological results from labellar chemoreceptors indicate that the previously reported hydrocarbon behavioral rejection thresholds are best explained by the inhibition of the sugar receptors. This conclusion is strengthened by further electrophysiological and behavioral tests on tarsal chemoreceptors.     

二点委夜蛾卵巢发育分级及在预测预报中的应用

对室内饲养的二点委夜蛾Athelis lepigone(Moschler)雌成虫分时段进行解剖,观察其卵巢的结构及发育进程。结果显示二点委夜蛾具有1对卵巢,各由4个卵巢小管组成。发育进程可分为5个阶段:透明期、卵黄沉积期、成熟待产期、产卵盛期和产卵末期。河北省石家庄地区2011年7月下旬至8月上旬田间卵巢发育的监测结果显示,Ⅰ级卵巢在整个发生期所占比例较高,Ⅳ级卵巢所占比例较低,推测其下一代幼虫的发生量将较低,并与田间调查结果吻合,因此卵巢解剖分级法可以用于二点委夜蛾的预测预报工作。     

Sugar response differences related to sensillum type and location on the labella of Protophormia terraenovae: a contribution to spatial representation of the stimulus

The chemoreceptor spike activity in response to sucrose in the concentration range 1-500mM was recorded from each of the 11 Intermediate and 13 Large labellar sensilla in the blowfly Protophormia terraenovae. The results showed that: (1) three of the four cells present in each sensillum are activated by sucrose stimulation; (2) differences between the Large and Intermediate types exist in the dose-response profiles of one of these cells (the 'water' cell), possibly reflecting different sugar receptor site populations on the dendritic membranes of homologous cells in the two types; (3) sensilla of both types are differentially responsive to sucrose solutions according to their location on the labellum. These differences may provide elements for a spatial representation of the stimulus source within the sensory coding process.     

Clonidine effects on protein and carbohydrate electrophysiological responses of labellar and tarsal sensilla in Phormia regina

The electrophysiological response of labellar and tarsal chemosensilla in the blowfly Phormia regina was studied in response to a complex stimulus naturally encountered by flies such as sheep faeces, and to beef liver, a proteinaceous feeding source. Responses were investigated both before or after injection of clonidine, an octopamine agonist previously shown to enhance sucrose ingestion, while decreasing that of proteins. As assessed by single sensillum recordings, the four different chemosensory - "salt", "sugar", "deterrent" and "water" - cells were all activated by both stimuli, regardless of sex and sensillum type, the "sugar" one being in all cases the most sensitive to beef liver before clonidine injection. Clonidine treatment affected neither labellar nor tarsal sensitivity to sucrose. Conversely, clonidine-injected flies showed a significant increase in the activity of the "deterrent" cell to beef liver, thus accounting for a decrease in protein ingestion. This study for the first time provides evidence of a key role of a clonidine-sensitive peripheral taste sensitivity in down-regulation of protein ingestion in blowflies. Correlation between peripheral sensitivity and behavioural output is discussed.     

The sensitivity of the labellar sugar receptors of Phormia regina in relation to feeding

The hypothesis that behavioural feeding threshold of P. regina modulates and is modulated by the sensitivity of the labellar contact chemoreceptors is not supported by the examination of individual flies.Sensitivity (number of impulses fired in the first second of stimulation) of the sugar receptors in selected labellar hairs of flies of known age was recorded periodically on a fixed schedule for up to 72 hr. Experimental flies were hand fed 2.0 M fructose once every 24 hr. No correlations between feeding or age and receptor sensitivity were found. Statistically significant changes in firing frequency did occur but were unrelated to feeding and age.The proboscis extension response, on which behavioural threshold measures are based, is known to be triggered by the first few sugar receptor spikes of sufficient frequency. Neither age nor feeding significantly affected the number or frequency of impulses during the first 50 msec of stimulation.     

The Drosophila seminal fluid protein Acp26Aa stimulates release of oocytes by the ovary

Mating stimulates the rate of egg-laying by female insects. In Drosophila melanogaster this stimulation is initially caused by seminal fluid molecules transferred from the male (Acps or accessory gland proteins; reviewed in [1] [2] [3]). Egg-laying is a multi-step process. It begins with oocyte release by the ovaries, followed by egg movement down the oviducts and the deposition of eggs onto the substratum. Although two Acps are known to stimulate egg-laying [4] [5], they were detected by assays that do not discriminate between the steps of this process or allow examination of its earliest changes [4] [5] [6] [7]. To determine how egg-laying is regulated, we developed a generally applicable assay to separate the process into quantifiable steps, allowing us to assess the ovulation pattern and rate of egg movement. As the steps are interdependent yet potentially subject to independent controls, we determined the contribution of each step and effector independent of the others. We used a statistical method [8] [9] that separately considers and quantifies each 'path' to a common end. We found that the prohormone-like molecule Acp26Aa [5] [10] stimulates the first step in egg-laying - release of oocytes by the ovary. During mating, Acp26Aa begins to accumulate at the base of the ovaries, a position consistent with action on the ovarian musculature to mediate oocyte release. Understanding how individual Acps regulate egg-laying in fruitflies will help provide a full molecular picture of insects' prodigious fertility, of reproductive hormones, and of the roles of these rapidly evolving proteins [11] [12].     

The Effects of Amino Acids on the Labellar Hair Chemosensory Cells of the Fly

The effects of amino acids on the labellar hair chemosensory cells were examined with two kinds of flies (the fleshfly, Boettcherisca peregrina, and the blowfly, Phormia regina). As a result of this examination, the effects of amino acids were divided into four main classes. Amino acids in class 1 did not stimulate any chemoreceptor cell. Amino acids in class 2 inhibited nonspecifically the discharges from three kinds of chemosensory cells. Amino acids in class 3 stimulated the salt receptor cell. Amino acids in class 4 stimulated the sugar receptor cell. A possibility that a fourth neuron in the labellar hair chemosensory cell might be a protein or an amino acid receptor cell was eliminated.     

Sensitivity of the mosquito Aedes aegypti (Culicidae) labral apical chemoreceptors to phagostimulants

The feeding of Aedes aegypti (L.) on blood and nectar is induced by phagostimulants: adenine nucleotides and sugars respectively. This work examines the responses of the four chemoreceptor cells in the labral apical sensilla to these phagostimulants. The apical chemoreceptors can detect the presence of adenine nucleotides. This part of the response is in good agreement with the gorging behavior. The output of the chemoreceptors cannot distinguish among different adenine nucleotides or among their concentrations (0.01-1 mmol/l), whereas gorging behavior is affected by the identity of adenine nucleotides and by their concentrations. Hence the gorging behavior cannot be driven by the output of these chemoreceptors alone. To the presence of adenine nucleotides Cell 2 was the only cell that responded with high frequencies, while the response of Cell 4 was almost abolished. The response of Cell 2 to ATP depended on the mosquito's physiological state. This dependence accorded well with the gorging behavior; Cell 2 responded with a higher frequency to ATP in the gorging state, than when not in a gorging state. The responses to sucrose and fructose constituted the only case recorded in which all these chemoreceptors failed to respond. This depression of response implies that other chemoreceptors must be present as sugar detectors.     

Peripheral chemoreceptors in health and disease.

Peripheral chemoreceptors (carotid and aortic bodies) detect changes in arterial blood oxygen and initiate reflexes that are important for maintaining homeostasis during hypoxemia. This mini-review summarizes the importance of peripheral chemoreceptor reflexes in various physiological and pathophysiological conditions. Carotid bodies are important for eliciting hypoxic ventilatory stimulation in humans and in experimental animals. In the absence of carotid bodies, compensatory upregulation of aortic bodies as well as other chemoreceptors contributes to the hypoxic ventilatory response. Peripheral chemoreceptors are critical for ventilatory acclimatization at high altitude. They also contribute in part to the exercise-induced hyperventilation, especially with submaximal and heavy exercise. During pregnancy, hypoxic ventilatory sensitivity increases, perhaps due to the actions of estrogen and progesterone on chemoreceptors. Augmented peripheral chemoreceptors have been implicated in early stages of recurrent apneas, congestive heart failure, and certain forms of hypertension. It is likely that chemoreceptors tend to maintain oxygen homeostasis and act as a defense mechanism to prevent the progression of the morbidity associated with these diseases. Experimental models of recurrent apneas, congestive heart failure, and hypertension offer excellent opportunities to unravel the cellular mechanisms associated with altered chemoreceptor function.     

Flower abscission and inflorescence carbohydrates in sensitive and non-sensitive cultivars of grapevine

The Gewurztraminer (GW) and the Pinot noir (PN) cultivars of grapevine differ in their sensitivity to environmental factors that can cause flower abscission, cv. GW being the most sensitive. In order to further define the mechanisms leading to abscission, and owing to the importance of sugars in the achievement of sexual organ ontogenesis, we attempted to correlate the chronology of flower ontogenesis with the variations of carbohydrates in the inflorescence. In the vineyard, under optimal climatic conditions, fruit set of cv. GW and cv. PN was 82% and 65%, respectively. The sugar distribution was different in their inflorescences during the entire duration of flower development. Between stages 15 and 17, flowers of GW and PN reached the crucial meiosis stage. At that time, the inflorescences of cv. GW exhibited higher concentrations of starch and sucrose, whereas those of PN presented higher levels of glucose and fructose. Despite higher starch concentrations in GW inflorescences, starch reserves were present in the ovules and anthers of PN but not in those of GW. These results suggest that the higher content of reserve and transport carbohydrates in the inflorescences of GW favour flower development and fruit set under optimal environmental conditions. Furthermore, since meiosis represents a key step of female development, the different sugar concentrations in the inflorescences of the two cultivars at stages 15 and 17 could be related to the sensitivity to flower abscission under climatic stress. In particular, the presence of starch granules in PN ovules and anthers might explain the higher resistance of this cultivar to flower abscission.     

Biochemical and physiological evidence that calmodulin is involved in the taste response of the sugar receptor cells of the blowfly, Phormia regina

The gustatory system is essential for almost all animals. However, the signal transduction mechanisms have not yet been fully elucidated. We isolated labellar chemosensilla from blowfly, Phormia regina, and purified calcium binding proteins from the water soluble fraction. The most abundant calcium-binding protein was calmodulin. To investigate the role of calmodulin in taste transduction, electrophysiological responses were recorded with the calmodulin inhibitor, W-7. When we stimulated the labellar chemosensillum with sucrose plus W-7, a dose-dependent decrease of impulse frequency was observed when the concentration was <50 microM. In addition, when W-7 at 50 microM or higher concentration was added, an initial short-term impulse generation from the sugar receptor cell was observed, but this was followed by a silent period. When the sensillum was stimulated with W-7 plus a membrane-permeable cGMP analog, dibtyryl-cGMP or 8-bromo-cGMP, impulses of the sugar receptor cell were induced but the frequency was decreased. By the sidewall-recording method, we observed that the receptor potential induced by sucrose stimulation was decreased by W-7 in the sugar receptor cell, and corresponded with a disappearance of impulses. These data strongly suggest that the cGMP-gated channel generating receptor potential in the sugar receptor cell requires calmodulin for its gating.     

Effects of alkaloids on feeding by Phormia reginaconfirm the critical role of sensory inhibition

Abstract. Inhibition of peripheral chemoreceptors by micromolar concentrations of alkaloids may account for feeding deterrence. Earlier work showed a reduction in both the proboscis extension response and sensory activity recorded from sugar-sensitive cells in tarsal D and labellar 'largest' hairs when the stimulating solution was sucrose mixed with alkaloids. In the present work a similar behavioural effect was also seen when alkaloids were mixed with pyranose and furanose sugars as well as with water and deuterium oxide. Behavioural deterrence continued after the stimulus was removed. Complete recovery occurred after 120 s. To check against the possibility of a central inhibitory state (CIS) having been established two sets of experiments were run in which quinine was applied to a single tarsus followed by a sucrose post-test to the contralateral tarsus. In one half of the tests sugar was applied first (S₁Q₂), and in the other half quinine first (Q₁S₂). Comparable tests were run with contralateral labellar hairs. No CIS was set up by Q₁; however, a central excitatory state (CES) was set up by S₁. In order to test (a) whether quinine had different effects on other known or unknown receptors and (b) post-ingestive effects, volumes imbibed in single drinks of sucrose with and without quinine were compared. Volumes of the mixture imbibed were not significantly different from what was predicted based on the known reduction in sensory acitivty.     

Systems of chemoperception in Decapod crayfish

The review presents data on some peripheral and central structures in the system of perception of chemical stimuli in crayfish and other Decapoda. The hair receptors on chelipeds, antennas, and antennules are innervated by mechano- and chemoreceptor neurons. Antennules are crayfish specialized chemoreceptor organs whose surface contains groups of exteroceptors. On claws of ambulatory feet (AF), antennas, antennules, and other mobile appendages there is a regular disposition of exteroceptor receptive fields in the form of receptor hair bushes. Behavioral experiments have shown sensitivity of crayfish to odor of individuals of their gender and sex partners as well as the presence in crayfish of pheromones providing connection of female with offspring at the initial stages of their life cycle. Individual chemosensory cells innervating hair bushes on the crayfish AF respond to amino acids, amines, nucleotides, and sugars. Minimal thresholds of reaction of the studied Decapoda chemoreceptors in response to some chemical compounds correspond to 0.1–1.0 μM. For some chemoreceptors, dose-dependent effects have been shown. Alongside with monomodal chemoreceptors, the crayfish have bimodal receptors perceiving mechanical and chemical stimuli. The efficient response of crayfish chemoreceptors can be obtained to the substance that has amino group with hydrogen bridge to carboxyl group, contains no more that 3 carbon atoms in the chain, is characterized by a certain stereoform. Among chemoreceptors there are fast and slow adapting cells. Efficiency of response of individual chemoreceptors depends on temperature of medium. In crayfish, chemoreceptors responding to ecdysterons have been revealed. Ecdysterons play a great role in intra- and inter-species communications in Crustacea. Based on the study of efferent responses of interneurons of the first and higher orders in the first thoracic crayfish ganglion to stimulation of the own receptive fields, a concept has been put forward of the structural-functional organization of afferent projections at the segmental level. Peculiarities of afferent projections from antennule chemoreceptors are considered. The data are presented on connections of these chemoreceptors with antennular, olfactory, and accessory lobules, various cell groups, interneurons of the first, second, and third orders located in various brain parts. An attention has been drawn to connections of serotonin neurons in glomeruli with endings of chemoreceptor neurons and projections of interneurons of the higher orders, which are located in the internal medulla of the crayfish eyestalks. Several principles of integration of the chemoreceptor information in central parts of the crayfish nervous system are discussed. The giant serotonin neurons revealed in crayfish glomeruli most likely participate in formation of memory to certain chemical actions. Polymodal receptor signals in the central chain of the perception system activate autonomic centers, and the changes of the animal functional state can be evaluated from the heart responses. These responses recorded by novel noninvasive methods allow detection of the initial and other phases of the stress state at changes of the medium chemical quality. Progress of the current biochemical and electrophysiological methods of study chemoreceptors allow hoping for learning of fine chemoperception mechanisms in invertebrate and vertebrate animals.     

Clustering of the Chemoreceptor Complex in Escherichia coli Is Independent of the Methyltransferase CheR and the Methylesterase CheB

The Escherichia coli chemoreceptors and their associated cytoplasmic proteins, CheA and CheW, cluster predominantly at the cell poles. The nature of the clustering remains a mystery. Recent studies suggest that CheR binding to and/or methylation of the chemoreceptors may play a role in chemoreceptor complex aggregation. In this study, we examined the intracellular distribution of the chemoreceptors by immunoelectron microscopy in strains lacking either the methyltransferase CheR or the methylesterase CheB. The localization data revealed that, in vivo, aggregation of the chemoreceptor complex was independent of either CheR or CheB.     

The use of isothermal titration calorimetry to unravel chemotactic signalling mechanisms

Chemotaxis is based on the action of chemosensory pathways and is typically initiated by the recognition of chemoeffectors at chemoreceptor ligand-binding domains (LBD). Chemosensory signalling is highly complex; aspect that is not only reflected in the intricate interaction between many signalling proteins but also in the fact that bacteria frequently possess multiple chemosensory pathways and often a large number of chemoreceptors, which are mostly of unknown function. We review here the usefulness of isothermal titration calorimetry (ITC) to study this complexity. ITC is the gold standard for studying binding processes due to its precision and sensitivity, as well as its capability to determine simultaneously the association equilibrium constant, enthalpy change and stoichiometry of binding. There is now evidence that members of all major LBD families can be produced as individual recombinant proteins that maintain their ligand-binding properties. High-throughput screening of these proteins using thermal shift assays offer interesting initial information on chemoreceptor ligands, providing the basis for microcalorimetric analyses and microbiological experimentation. ITC has permitted the identification and characterization of many chemoreceptors with novel specificities. This ITC-based approach can also be used to identify signal molecules that stimulate members of other families of sensor proteins.     

Effects of mating, nutritional condition, and mouthpart separation on ingestion and destination of sugar and water by the mosquito Culiseta inornata

ABSTRACT. 'Hungry' tethered female Culiseta inornata (Williston) (Diptera: Culicidae) with their mouthparts separated, respond in the sugar-feeding mode when the fascicle is immersed in 1 M sucrose, and in the drinking mode when the fascicle is immersed in water. Similarly treated animals that are 'not hungry' respond in the drinking mode to water on the fascicle, but when given 1 M sucrose on the fascicle take less than a normal amount and tend to put it in the midgut instead of the crop. 'Starving' animals respond in the sugar-feeding mode to sucrose on the fascicle, but when given water on the fascicle take slightly more than a normal amount, and about 50% put it in the crop. These results occurred whether the labellum was immersed in water, 1 M sucrose, or nothing. Differences in responses by tethered animals with separated mouthparts and non-tethered intact animals can be attributed to mouthpart deployment. These results do not agree with those of other workers who report that similarly treated female C. inornata require stimulation of both the fascicle and labellar chemoreceptors before they will ingest. These differing results raise interesting questions concerning the range of feeding behaviours available to different demes of this mosquito, and indicate how these behaviours are modified by differences in pre-test nutritional condition and mouthpart deployment.     

瓜实蝇卵巢的发育阶段及分级

瓜实蝇是热带和亚热带地区最重要的经济害虫之一。本研究拟对卵巢发育进行分级,以判断田间捕获的瓜实蝇是否达到性成熟。室内饲养瓜实蝇成虫,对开始羽化至羽化后80 d的不同日龄成虫卵巢进行解剖,并记录了卵巢长度、卵巢宽度、卵巢指数（卵巢长度＆#215;卵巢宽度）和所含卵粒数。将瓜实蝇卵巢发育过程分为4期6个级别,分别为卵黄发生前期（Ⅰ级和Ⅱ级）、卵黄发生期（Ⅲ级和Ⅳ级）、抱卵期（Ⅴ级）、经产期（Ⅵ级）。随着卵巢的发育,Ⅰ～Ⅴ级卵巢长度和宽度迅速增长,卵巢发育成熟开始产卵以后,Ⅵ级卵巢长度和宽度逐渐减小。Ⅰ级和Ⅱ级卵巢长度与宽度大致相等,以后各期卵巢长度均大于宽度。Ⅲ级和Ⅳ级可见清晰的卵室,卵母细胞迅速增长,卵黄逐渐沉积。Ⅴ级可见清晰的卵粒,整个卵巢呈圆柱状。Ⅵ级开始排出卵子,所含卵粒数显著少于Ⅴ级;虽然此级卵巢长度大于Ⅳ级,但卵巢宽度、卵巢指数与Ⅳ级基本一致;Ⅵ级卵巢可见黄体,可结合黄体判定卵巢所处发育级别。本研究对评估粘虫板捕获的瓜实蝇雌虫生理日龄结构有潜在应用价值。     

棉铃虫卵巢形态与卵子发生过程观察

害虫发生高峰期、 发生量的准确预测和田间防治适期的确定与种群雌虫卵巢结构及卵子发生过程密切相关。为了明确棉铃虫Helicoverpa armigera卵巢结构及卵子发生过程, 本研究利用光学体视显微镜和透射电子显微镜, 对棉铃虫成虫卵巢管和卵子的超微结构进行了研究, 并确定了发育级别划分标准。结果表明: 根据卵巢的形状、 卵的产生过程、 卵黄沉积情况等将棉铃虫卵巢发育程度分为6个级别, 即发育初期(0级)、 卵黄沉积前期(Ⅰ级)、 卵黄沉积期(Ⅱ级)、 成熟待产期(Ⅲ级)、 产卵盛期(Ⅳ级)和产卵末期(Ⅴ级)。根据卵子发生过程中滋养细胞、 卵母细胞的变化, 将卵子发生期分为3个阶段： 卵黄发生前期、 卵黄发生期和卵黄成熟期。本研究首次对棉铃虫的卵子发生进行电子显微观察, 并完善了棉铃虫卵巢发育的分级标准, 为进一步研究棉铃虫的生殖发育机理提供了理论参考, 对田间棉铃虫种群发生期和发生量的预测预报也有重要的实践参考价值。     

不同地区田间草地贪夜蛾卵巢的结构和发育

【目的】探明田间草地贪夜蛾Spodoptera frugiperda卵巢的发育状态及其变化,为草地贪夜蛾种群的发生和迁飞动态提供精准监测依据。【方法】采用雌性生殖系统解剖法,比较不同日龄和地区的田间草地贪夜蛾雌蛾卵巢的形态差异,分析卵巢成熟状态与雌蛾交配的关系。【结果】草地贪夜蛾雌蛾生殖系统包含卵巢、输卵管、交配囊、导精管、受精囊、附腺和产卵器。根据形态特征可以将田间草地贪夜蛾卵细胞分为卵黄发生前期、卵黄期和成熟期3个时期。田间雌蛾1日龄卵巢中发育最快的卵仅处于卵黄期中期,卵巢管柄为空;3日龄卵巢出现分化,一部分卵巢仍然和1日龄相似,另一部分卵巢中存在成熟卵;这种卵巢发育分化到羽化后11 d时仍然存在。云南江城、弥勒,广西田阳以及浙江瑞安和镇海等地的田间雌蛾,到死亡时分别有61.5%, 51.7%, 41.7%, 42.1%和35.5%卵巢发育不成熟。室内饲养第1代雌蛾中,有39.6%的个体到死亡时卵巢未发育成熟,和田间雌蛾比例相似。交配雌蛾卵巢可以是发育成熟和未成熟的,但是未成熟卵巢的比例仅为18.0%±5.0%。【结论】结果提示同一代次的田间草地贪夜蛾可能同时存在迁飞和非迁飞个体,其比例随地理位置和季节发生变化。本研究的结果在一定程度上解释了单纯依据化学农药难以防治草地贪夜蛾的原因,也为田间草地贪夜蛾的迁飞监测和绿色防控提供了一定依据。