Age-related changes in the response to adipokinetic hormone in Locusta migratoria

ABSTRACT. In fifth instar nymphs of Locusta there is only a feeble adipokinetic response to extracts of corpora cardiaca. In fledglings, this poor response persists for a few days but then increases dramatically to reach a plateau by day 8. The response declines to almost zero as the locusts age beyond 35 days of adult life. This pattern of change in response is similar in both males and females but there are some differences in magnitude depending upon whether the response is measured as changes in haemolymph total lipid (vanillin-positive material) or total diglyceride (gas liquid chromatography). The poor response to adipokinetic hormone in nymphs and newly fledged locusts is not a result of shortage of stored lipid in the fat body and cannot be improved by injection of extra hormone.     

Measurements of locomotor activity in hatchlings of the migratory locust Locusta migratoria: effects of intrinsic and extrinsic factors

Abstract. The effects of intrinsic and extrinsic factors on locomotor activity in crowd‐reared first‐stadium nymphs of the migratory locust Locusta migratoria are investigated under continuous light conditions using an actograph apparatus. Nymphs show monomodal or bimodal patterns of locomotor activity with respect to the time after the start of measurements, depending on the age. Locomotor activity is suppressed by the presence of grass in nymphs aged 0–2 days old but a peak of activity observed shortly after hatching is not suppressed. The results suggest that newly‐hatched nymphs may try to disperse from the hatching site. Nymphs show higher locomotor activity levels under moist conditions than under dry conditions during the first 5‐h period of measurements. This enhanced locomotor activity may constitute attempts to avoid high humidity. Under dim‐light conditions (2 × 10^?2 Wm^?2), locomotor activity is suppressed during the first half day and increases to a high level thereafter in both grass‐fed and unfed individuals. This increased activity might indicate a possible involvement of circadian rhythms. Background colour has no significant effect on the locomotor activity. The present study provides new aspects of behaviour in nymphs as well as baseline data for behavioural analysis of locust locomotion in relation to phase polyphenism.     

Changes in chemoreceptor sensitivity in relation to dietary selection by adult Locusta migratoria

During somatic growth in adult Locusta migratoria L. there are variations in the responsiveness of chemosensilla on the maxillary palps. Relative changes in chemosensitivity to amino acids and sucrose are consistent with relative changes in protein and carbohydrate ingestion. There is a decline throughout the somatic growth phase in the proportion of sensilla responding to stimulation. In those sensilla which respond, there is a decline in the firing rate to amino acids, while sucrose responsiveness declines to a lesser degree. We suggest that variations in the levels of nutrients in the storage tissues, especially the haemolymph, arising from changes in the demand for protein and carbohydrate for growth, directly or indirectly, influence gustatory sensitivity.

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Résumé Pendant la croissance de L. migratoria L. les réactions aux acides aminés et aux sucres des sensilles chimiques des palpes maxillaires changent. Ces changements relatifs de la sensibilité chimique à ces deux aliments correspondent aux modifications dans l'ingestion de protéines et d'hydrates de carbone par cet insecte. La diminution de la réponse aux acides aminés est liée à la fois au nombre de sensilles qui réagissent et à leur taux de réaction. La sensibilité des sensilles au sucrose varie moins profondément et implique uniquement des modifications du nombre de sensilles réagissant. Nous suggérons que des variations de la teneur en éléments nutritifs de l'haemolymphe, dues à des modifications de la demande en protéines et en glucides nécessaires à la croissance, influent sur la sensibilité gustative.

     

Changes in haemolymph osmotic pressure in Locusta migratoria larvae in relation to feeding

An account is given of the effect of feeding on haemolymph osmotic pressure in larvae of Locusta migratoria. Changes in concentration of solutes and haemolymph volume are investigated and discussed.     

飞蝗越冬卵过冷却点的季节性变化及生态学意义

运用热电偶方法测定了飞蝗LocustamigratoriaL .越冬卵过冷却点月份间的变化。结果表明 :飞蝗卵的过冷却点值波动较大 ,主要形成 2部分 ,一部分分布在 -2 5℃左右 ,另一部分分布在 -1 0℃左右 ,同时 ,这两部分的比例随着季节性气温的变化有明显的差异 ,气温较低的月份 ,-2 5℃左右的比例增加 ,而且最小过冷却点的值也更低 ;反之 ,-1 0℃左右的比例增加 ,过冷却点最高值也升高。这种现象说明了飞蝗卵对冬季低温的一种生态适应。     

Behavioural differences in Locusta migratoria associated with albinism and their relation to [His7]-corazonin

Abstract. The neuropeptide [His⁷]-corazonin induces black colouration in locusts, a feature typically occurring in gregarious animals. To date, the function of this neuropeptide in relation to phase transition has not been fully clarified. Unanswered questions on its involvement in behavioural and morphometrical phaseshifting and possible temperature-induced regulation still remain. In the African locust, Locusta migratoria migratorioides (Reiche and Fairmaire) (Orthoptera: Acrididae), the gregarious form possesses mainly a brown colouration with typical intense black pigmentation. To find a possible behavioural function, an albino L. migratoria mutant, deficient in [His⁷]-corazonin and thus lacking the typical gregarious colour, was used. Between these two phenotypes, nonphase specific behavioural differences in a phase-dependent behavioural set-up were found. Additionally, a wild-type of the Okinawa strain was screened behaviourally and located in comparison to the other groups. Treatment of albinos with corazonin induced normal phenotype behaviour. The degree of interference of corazonin in relation to these findings and a possible effect at the level of visual perception is discussed.     

Developmental expression of odorant‐binding proteins and chemosensory proteins in the embryos of Locusta migratoria

We have investigated the development of chemosensilla and the secretion of odorant‐binding proteins (OBPs) and chemosensory proteins (CSPs) in the embryo of Locusta migratoria manilensis. We first report the changes of each sensillum in embryo just preceding hatch in detail and show that different sensilla have different developmental processes. Trichogen cells are first involved in forming the structure of pegs, and then, after retraction, they start secreting OBPs and CSPs in the sensillar lymph. The synthesis of LmigOBP1 starts during the embryogenesis about 0.5 h preceding hatching, specifically in sensilla trichodea and basiconica of the antenna. LmigOBP2, instead, was only found in the outer sensillum lymph (oSl) of sensilla chaetica of the antenna, while we could not detect LmigOBP3 in any type of sensilla of the antenna. The ontogenesis of CSPs in the embryos is similar to that of OBPs. Expression of CSPI homolog in Locusta migratoria is detected using the antiserum raised against SgreCSPI. CSPI is specifically expressed in the outer sensillum lymph of sensilla chaetica of the antenna, and anti‐LmigCSPII dose not label any sensilla of the embryos. These data indicate that in locusts, OBPs and CSPs follow different temporal expression patterns, and also that OBPs are expressed in different types of sensilla. © 2009 Wiley Periodicals, Inc.     

The structure of the terminal sensilla on the maxillary palps of Locusta migratoria (L.), and changes associated with moulting

Summary The basic structure of the terminal sensilla of Locusta migratoria resembles that of Schistocerca gregaria. There are commonly six or ten neurons whose dendrites extend almost to the opening of the peg. Proximally the dendrites are clothed by a neurilemma cell which also encloses a basal cavity through which their ciliary region passes. The tormogen cell encloses the receptor-lymph cavity and actively secretes material into it. The receptor-lymph cavity and the basal cavity are quite separate.The development of new pegs at a moult is described. After apolysis the scolopale extends across the subcuticular space and protects the dendrites, which remain in a functional condition until shortly before ecdysis. As the trichogen cell grows out to form a new peg the tip is surrounded by a mass of electron-dense material, probably derived from the receptorlymph cavity. The function of this material is unknown. Regeneration of the dendrites is considered.The possible mechanism by which the tip of the peg opens and closes is considered and the general structure of the organule is discussed in relation to functioning.     

Developmental changes of the glycolytic enzymes in the human fetal heart

The ontogeny of hexokinase, phosphofructokinase, phosphoglucoisornerase, aldolase, pyruvate kinase and lactate dehydrogenase activities which are associated with glycolysis, an important energy yielding process, has been studied in human fetal heart for periods ranging from 13 weeks to above 33 weeks of gestation. Hexokinase, phosphoglucoisomerase and pyruvate kinase activities show similar developmental profiles exhibiting maximum activity at 25–28 weeks ofgestation. Phosphofructokinase activity, on the other hand, shows a minimum at this period and exhibits a peak value at early stages (13–16 weeks of gestation). Though considerable activity for aldolase is observed at an early period, it declines thereafter, but again increases in the later period. The probable role and correlations of these glycolytic enzymes with energy demand and general functional development in human fetal heart in ontogeny are evaluated.     

Developmental changes in ventricular myosin isoenzymes of the tammar wallaby

Ventricular myosin in eutherian mammals undergoes a perinatal change in response to a sharp rise in thyroid hormone levels during development. In this investigation, changes in ventricular myosin heavy chains (MyHCs) of the tammar wallaby (Macropus eugenii) from early pouch life to adulthood were analysed using native gel electrophoresis, SDS-PAGE and western blotting. Adult wallaby ventricle showed three myosin isoenzymes, V₁, V₂ and V₃; western blots using specific anti-α-MyHC and anti-β-MyHC antibodies showed their MyHC compositions to be αα, αβ and ββ, respectively. Ventricular muscle in early pouch joeys expressed predominantly β-MyHC. Up to 200 days, the time of initial pouch exit, α-MyHC content was around 5%. Thereafter, there was a sharp increase of α-MyHC expression to 35% by 242 days of age, eventually falling back to 23% in the adult. These changes correlate with known surges in plasma levels of thyroid hormones around pouch exit. The results suggest that ventricular myosins in a marsupial mammal also undergo a developmental change, and that marsupial ventricular myosins are thyroid responsive as in eutherians. The increased α-MyHC expression empowers the heart to meet the enhanced cardiovascular demands of out-of-pouch activity and the thermogenic action of thyroid hormones.     

Carbohydrases and carbohydrate digestion in the gut of Locusta migratoria

The digestion of various carbohydrates and synthetic substrates by the gut of Locusta migratoria was analysed quantitatively. Maltose, starch, and sucrose were found to be hydrolysed most rapidly, whereas the splitting of cellobiose, trehalose, lactose, and melecitose took place at much slower rates.The absolute carbohydrase activities in foregut and midgut are nearly equal. However, specific activities are much higher in the foregut. Only low activities were found in extracts from the hindgut and salivary glands. The latter show a pattern of sugar splitting which is different from that found in gut preparations.The distribution of carbohydrase activities between the epithelia and lumina of the foregut, midgut, and hindgut and between soluble and particulate fractions were studied. The midgut epithelium is shown to have a particularly high content of enzymes, although some carbohydrases are rather active also in the epithelium of the hindgut. During hunger periods the relative enzymatic activities of the epithelium are distinctly increased.The isolation and purification of the carbohydrases were attempted and a partial separation of individual enzymes was obtained by gel-filtration. These results indicate the presence of at least seven distinct carbohydrases in the locust gut. The molecular weights of the enzymes were estimated by gel-filtration, and K_M values and pH-optima are reported.     

东亚飞蝗主要过敏原的分析、鉴定与纯化

通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离东亚飞蝗Locusta migratoria manilensis(Meyen)的蛋白质组分并测定其分子量,收集过敏病人血清,采用免疫印迹(Western—blotting)法鉴定其过敏原成分,通过凝胶过滤层析对东亚飞蝗过敏原进行分离纯化。结果表明:东亚飞蝗蛋白粗提液条带大概有30条左右,其中主带大约有10条,相对分子量约为13、15、25、28、40、45、55、70、100、110ku,其中蛋白含量最丰富的约在70ku左右。免疫印迹结果显示,蝗虫过敏条带主要有5条,相对分子量分别约为19、29、38、70、130ku。通过凝胶过滤层析对东亚飞蝗过敏原进行分离纯化,得到了一个高纯度相对分子质量约为70ku东亚飞蝗过敏原,并且发现了一个相对分子质量约为130ku的蝗虫新过敏原。本研究为临床上蝗虫食物变态反应性疾病的诊断和治疗奠定基础。     

Chemo-discriminatory neurones in the sub-oesophageal ganglion of Locusta migratoria

The sub-oesophageal ganglion of Locusta migratoria was searched for neurones responsive to stimulation of the maxillary palps using intracellular recording techniques. Two plant stimuli were used: wheat, a host plant and cabbage, an unacceptable non-host plant. The stimuli were presented to the palp as both intact leaf tissue and as droplets of aqueous solutions of plant extracts. All the sampled neurones that responded to stimulation of the palp also responded to simple mechanical stimulation. However, 25% of the neurones exhibited consistent differences in response to the two plants when presented as both leaf tissue and droplets, strongly suggesting that they also received a chemosensory input. These differential responses most commonly took the form of differences in the duration of cell activity and/or variation in the latency of the onset of response. The receptive fields of differentially responding neurones were confined to the maxillary palp, or at most to the maxillary and labial palps.     

Secretory granule formation and membrane recycling by the trans-Golgi network in adipokinetic cells of Locusta migratoria in relation to flight and rest

The influence of flight activity on the formation of secretory granules and the concomitant membrane recycling by the rans-Golgi network in the peptidergic neurosecretory adipokinetic cells of Locusta migratoria was investigated by means of ultrastructural morphometric methods. The patterns of labelling of the trans-Golgi network by the exogenous adsorptive endocytotic tracer wheat-germ agglutinin-conjugated horseradish peroxidase and by the endogenous marker enzyme acid phosphatase were used as parameters and were measured by an automatic image analysis system. The results show that endocytosed fragments of plasma membrane with bound peroxidase label were transported to the trans-Golgi network and used to build new secretory granules. The amounts of peroxidase and especially of acid phosphatase within the trans-Golgi network showed a strong tendency to be smaller in flight-stimulated cells than in non-stimulated cells. The amounts of acid phosphatase in the immature secretory granules originating from the trans-Golgi network were significantly smaller in stimulated cells. The number of immature secretory granules positive for acid phosphatase tended to be higher in stimulated cells. Thus, flight stimulation of adipokinetic cells for 1 h influences the functioning of the trans-Golgi network; this most probably results in a slight enhancement of the production of secretory granules by the trans-Golgi network.     

Octopamine modulates a central pattern generator associated with egg-laying in the locust,Locusta migratoria

Egg-laying in Locusta migratoria involves the control of a variety of complex behavioural patterns including those that regulate digging of the oviposition hole and retention of eggs during digging. These two behavioural patterns are under the control of central pattern generators (CPGs). The digging and egg-retention CPGs are coordinated and integrated with overlapping locations of neural substrate within the VIIth and VIIIth abdominal ganglia of the central nervous system (CNS). In fact, the egg-retention CPG of the VIIth abdominal ganglion is involved in both egg-retention and protraction of the abdomen during digging. The biogenic amine, octopamine, has peripheral effects on oviduct muscle, relaxing basal tension of the lateral and upper common oviduct and enabling egg passage. Here we show that octopamine also modulates the pattern of the egg-retention CPG by altering the motor pattern that controls the external ventral protractor of the VIIth abdominal segment. There is no change in the motor pattern that goes to the oviducts. Octopamine decreased the frequency of the largest amplitude action potential and decreased burst duration while leading to an increase in cycle duration and interburst interval. The effects of octopamine were greatly reduced in the presence of the α-adrenergic blocker, phentolamine, indicating that the action of octopamine was via a receptor. Thus, octopamine orchestrates events that can lead to oviposition, centrally inhibiting the digging behavior and peripherally relaxing the lateral and common oviducts to enable egg-laying.     

飞蝗羧酸酯酶基因LmCarE25的体外重组表达

羧酸酯酶(Carboxylesterase,CarE)是一类在生物中广泛分布的多功能家族酶系,在昆虫抗药性形成机制中发挥重要作用。为探讨飞蝗Locusta migratoria(Meyen)羧酸酯酶基因LmCarE25的生物学功能,本文采用荧光实时定量PCR技术进行研究,发现其在飞蝗成虫各组织部位均有表达,其中胃盲囊、翅和肌肉中表达量较低,中肠、马氏管和脂肪体表达量较高。本文尝试在大肠杆菌和昆虫细胞体外重组表达该酶,构建重组表达载体pET32a-LmCarE25,SDS-PAGE结果显示,LmCarE25可在大肠杆菌BL21(DE3)和JM109中表达,蛋白分子量约为60 ku,但为包涵体;将LmCarE25基因插入真核表达载体pFastBacHTA中,借助Bac-to-Bac体系获得重组Bacmid,以sf9细胞系作为宿主细胞表达目的蛋白,Western-Blot检测结果显示,LmCarE25获得可溶性表达。上述结果为进一步深入研究飞蝗羧酸酯酶家族功能提供了基础资料。     

Organ-specific distribution of the calcium sensor CaMKII in Locusta migratoria

The Ca(2+)/calmodulin-dependent kinase CaMKII is a key signaling component in Ca(2+)-dependent physiological processes. The expression and function of CaMKII in insect brain is well documented but less investigated for other tissues of insects. The present study demonstrates that in the locust Locusta migratoria CaMKII is widely expressed in various tissues. Relatively high expression levels of CaMKII were found in the brain, upper part of the digestive tract (pharynx, esophagus), and the flight and leg muscles. The different expression patterns of CaMKII in various tissues, as well as different molecular masses of CaMKII between 48 and 60 kDa indicate a tissue-specific expression of CaMKII variants. The expression was monitored with a polyclonal anti-(rat)CaMKII antibody. About 60% of total CaMKII activity in flight muscle cells is associated to the myofibril-rich, particulate fraction suggesting an important role of CaMKII in sarcomeric function.     

Noise filtering in the auditory system of Locusta migratoria L

Many acoustically communicating grasshoppers live in crowded populations where sound of many individuals may cause permanent noise. Tympanic receptors and first-order auditory interneurons of Locusta migratoria code such noise tonically, whereas many higher order interneurons react only weakly. In response to simultaneously presented sound they exhibit a better signal-to-noise ratio than their presynaptic elements. Two possible filter mechanisms are suggested for noise reduction in higher-order interneurons: (i) high-pass filtering of receptor spike frequencies and (ii) filtering due to synchronization of receptor spikes. Different receptor spike frequencies were elicited by series of short noise pulses with variable repetition rates. Receptor activities differing in their degree of synchronization were elicited by sound stimuli with variable rising times. In contrast to the first order interneurons some higher order interneurons responded best to receptor spike frequencies above 150–200 Hz, thus showing the postulated filtering. Only one higher order interneuron (AN4) distinguished between synchronous and asynchronous receptor activities. It is suggested that high-pass filtering of receptor spike frequencies is responsible for the noise filtering observed in these interneurons. The synchronization selectivity of AN4 is proposed to be responsible for temporal pattern detection of conspecific sounds.