Endocrine control of TAG lipase in the fat body of the migratory locust, Locusta migratoria

Aspects of the role and activation of the enzyme triacylglycerol lipase (TAG lipase) in the fat body of the migratory locust Locusta migratoria were investigated. TAG lipase is under the hormonal control of the three endogenous adipokinetic peptides of the migratory locust, Locmi-AKH-I, Locmi-AKH-II and Locmi-AKH-III. Injection of low doses (5-10 pmol) of each peptide causes an increase in lipase activity. The activation of lipase is time dependent: an elevated activity was recorded 15 min after injection of 10 pmol Locmi-AKH-I and maximum activation was reached after 45-60 min. The activation of TAG lipase is also dose-dependent. Doses of 2 pmol of each Locmi-AKH had no effect, whereas 5 pmol caused a significant activation. Maximum activation is reached with a dose of 10 pmol. Analogues of the second messengers cAMP (cpt-cAMP) and IP(3) (F-IP(3)) both activate the enzyme glycogen phosphorylase whereas only cpt-cAMP, but not F-IP(3), activates TAG lipase; cpt-cAMP elevates the lipid levels in the haemolymph. Activation of lipase is specific to the three endogenous AKH peptides: 5 pmol of the endogenous peptide Locmi-HrTH and 10 pmol of corazonin failed to activate lipase. High doses of octopamine did not activate lipase nor did they elevate the lipid concentration in the haemolymph. TAG lipase is stimulated by flight activity but activation is slower than that of glycogen phosphorylase: after 30 min of flight or after 5 min of flight plus 1h of subsequent rest, activity of TAG lipase is increased, but not immediately after 5 min of flight. In contrast, glycogen phosphorylase is activated significantly after 5 min of flight. These activation patterns of the two enzymes mirror-image the concentration of their substrates in the haemolymph: there is a significant decrease in the concentration of carbohydrates after 5 min of flight, whereas no change of the concentration of lipids can be measured after such short time of flight activity; however, a subsequent rest period of 1h is sufficient to increase the lipid concentration.     

飞蝗研究进展

飞蝗作为农业重大害虫,其分布遍及欧洲、亚洲、非洲和澳洲。全世界共有10个亚种,我国有3个亚种。飞蝗不但分布广,而且数量多、发生频率高,危害极为严重。作者在查阅了国内外近20年文献资料的基础上,结合我国蝗害情况,分别从形态、组织、生理、生化、遗传、毒力及分子系统学等方面综述了国内外飞蝗的研究进展,以期对我国飞蝗的基础及应用研究提供参考。     

Transport of d-galactose by the gastrointestinal tract of the locust, Locusta migratoria

Due to exoskeleton, the absorption of nutrients in adult insects takes place across the gastrointestinal tract epithelium. In most physiological studies, sugar intestinal absorption has been described as a diffusional process and to date no sugar transporter has been cloned from the digestive tract of insects. In the present work, the existence of a saturable transport system for galactose in the gastric caeca of Locusta migratoria is clearly demonstrated. This transport shows a relatively high affinity for galactose (apparent K0.5=2-3 mM) and is inhibited by glucose, 2-deoxyglucose and with less potency by fructose and alpha-methyl-d-glucoside. The absence of sodium or the presence of phloridzin hardly affects galactose absorption, indicating that it is not mediated by a SGLT1-like transporter. The absence of K+, Cl-, Mg2+ and Ca2+ or changes in the pH do not modify galactose absorption either. Nevertheless, phloretin, cytochalasin B and theophylline (inhibitors of facilitative transporters) decrease sugar uptake around 50%. Xenopus laevis oocytes microinjected with poly A+ RNA isolated from gastric caeca show sodium-independent galactose uptake that is three times higher than in non-injected oocytes, further supporting the existence of a mRNA coding for at least one equilibrative sugar transporter in L. migratoria gastric caeca.     

Neuroendocrine control of blood lipids in the locust, Locusta migratoria

In order to determine the best conditions, the influence of various parameters on the haemolymph lipid concentration were studied. These parameters are the age, the sex and the feeding of the animals, the time and the number of the haemolymph sample-taking and the temperature of the locust culture. A large in vivo increase in haemolymph lipid concentration was obtained in locusts which received extracts of the whole CC and of their glandular or neurohemal lobes. Reversely, a decrease in this concentration was obtained in locusts operated 7 days before (cardiacectomy or glandular lobe removal). Moreover the pars intercerebralis extracts increased the level of haemolymph lipids. We conclude that adipokinetic factors are present, both in the glandular lobes of the CC and in their neurohemal lobes. It is likely that the latter partly originate from the pars intercerebralis. Results of allatectomy and injections of corpora allata extracts led to the conclusion that corpora allata contain an adipokinetic factor, the juvenile hormone. and factors that inhibit the haemolymph lipid concentration. Finally, from different injections of neurotransmitters and drugs it is argued that it is mainly octopamine which is involved in the mechanism governing the increase of the level of haemolymph lipids.     

Characterization of cuticular proteins from the migratory locust,Locusta migratoria

Proteins were extracted from the still unhardened (teneral) cuticle of the migratory locust, Locusta migratoria. The proteins are soluble only at extreme pH-values and at low ionic strength, the solubility increases with decreasing temperature. The unhardened cuticle contains approx. 100 different proteins according to two-dimensional polyacrylamide gel electrophoresis. The majority of the proteins are very basic. The basicity and solubility properties of the proteins have necessitated development of modified electrophoretic procedures. The amino acid composition of the bulk protein shows that alanine, proline, glycine, valine and tyrosine constitute two thirds of the total amino acid content and that cysteine, methionine and tryptophan are absent.The proteins have been extracted from various parts of the cuticle and analysed by two-dimensional electrophoresis. Characteristic protein compositions were found for cuticle from the different body parts. Amino acid analyses of these extracts are strikingly similar. The only significant difference is in the glycine-alanine ratio. Cuticles that are destined to become hard are extremely rich in alanine, whereas the flexible parts of the cuticle are enriched in glycine. The results indicate that the proteins of locust cuticle constitute a group of structural proteins different from other known structural proteins.     

Hydrolysis of neutral glycerides by lipases of rat brain microsomes.

The hydrolysis of monoacylglycerol and diacylglycerol by rat brain microsomes was followed by measuring the release of glycerol and monooleylglycerol from dispersions of water insoluble glyceryl esters of oleic acid. The microsomes showed three lipolytic activities. One activity, optimal at pH 4.8, catalyzed the hydrolysis of diacylglycerol but not monoacylglycerol. Two other lipolytic activities, optimal at pH 8.0-8.6, catalyzed the hydrolysis of both diacylglycerol and monoacylglycerol. The pH 8.0-8.6 activities were sensitive to heat and SH-reagents. Detergents were inhibitory in all cases. Extraction of the microsomes with KCl, KSCN, urea or Triton X-100 did not change the ratio of diacylglycerol hydrolysis at pH 4.8 and 8.0. The results of subcellular fractionation studies showed that there was no significant enrichment of the acid lipase in any fraction.     

The neural control of spermathecal contractions in the locust,Locusta migratoria

The innervation of the spermatheca and demonstration of neural control of spermathecal contractions in Locusta migratoria was illustrated using anterograde and retrograde fills, combined with electrophysiological stimulation and recording. The anterior portion of the spermatheca receives innervation via the receptaculum seminis nerve (N2B2) from two large ventral neurons and one dorsal neuron. All were bilaterally paired and situated in the VIIIth abdominal ganglion. Three ventral bilaterally paired neurons situated in the VIIIth abdominal ganglion also provide innervation to the posterior portion of the spermatheca via the ductus seminalis aperture nerve (N2B3). Six DUM neurons, located in the VIIIth abdominal ganglion, in addition to two centroposteriorly situated DUM neurons in the VIIth abdominal ganglion, are also associated with these two nerves. N2B4 also provides innervation to the posterior portion of the spermatheca. N2B6b is associated with sensory cells identified in the anterior lateral regions of the genital chamber. The spermatheca contracts spontaneously, with peristaltic contractions beginning at the spermathecal sac and continuing along the length of the spermathecal duct. However electrical stimulation of the ventral ovipositor nerve (VON or N2B), receptaculum seminis nerve (N2B2) and the ductus seminalis aperture nerve (N2B3) indicates that contractions are also under neural control. In particular contractions of the spermathecal sac, coil duct and anterior straight duct are initiated via motor projections from the receptaculum seminis nerve (N2B2) and posterior straight duct contractions are controlled by motor input from the ductus seminalis aperture nerve (N2B3). The results suggest that spermathecal contractions of the anterior and posterior portions of the spermatheca are under separate neural control.     

沿海蝗区东亚飞蝗产卵选择的适应性

以沿海蝗区南大港水库为研究区域,通过2年野外调查旨在掌握东亚飞蝗Locustamigratoriamanilensis(Meyen)产卵场所的选择。结果表明,飞蝗产卵适宜的植被覆盖度、土壤含盐量和土壤5cm含水量分别为0~30%、0.09%~1.99%和10.0%~20.0%,当植被覆盖度>70%,土壤含水量>30%或含盐量>3%时,不再适合飞蝗产卵。土壤有机质和pH值不是影响飞蝗产卵选择的主要因素。与20世纪50年代的研究结果相比,飞蝗产卵时对土壤含水量和含盐量的耐受范围拓宽,对环境的适应能力增强。     

Deltamethrin-induced deregulation of the water balance in the migratory locust,Locusta migratoria

1. Several insecticides were tested for their ability to induce a water deregulation in the larval migratory locust. All of them provoked an accelerated dehydration (when compared to sham-operated insects). Deltamethrin and baygon were the most potent.2. This enhanced dehydration due to deltamethrin in adult locust resulted from an increase in the water loss through the feces. This increase was not due to a direct effect of deltamethrin on urine production by the Malpighian tubules but to a hormonal deregulation.3. Intoxicated insects produced large amounts of the vasopressin-like insect diuretic hormone. This higher synthesis activity occurs within the hours following the insecticide injection and is accompanied by an increase in water loss.4. These hormonal and metabolic modifications are transient. Hormonal level and diuresis rate both return to the basal levels 7 hr after the insecticide injection.     

Stress-induced thermotolerance of ventilatory motor pattern generation in the locust, Locusta migratoria

Ventilation is a crucial motor activity that provides organisms with an adequate circulation of respiratory gases. For animals that exist in harsh environments, an important goal is to protect ventilation under extreme conditions. Heat shock, anoxia, and cold shock are environmental stresses that have previously been shown to trigger protective responses. We used the locust to examine stress-induced thermotolerance by monitoring the ability of the central nervous system to generate ventilatory motor patterns during a subsequent heat exposure. Preparations from pre-stressed animals had an increased incidence of motor pattern recovery following heat-induced failure, however, prior stress did not alter the characteristics of the ventilatory motor pattern. During constant heat exposure at sub-lethal temperatures, we observed a protective effect of heat shock pre-treatment. Serotonin application had similar effects on motor patterns when compared to prior heat shock. These studies are consistent with previous studies that indicate prior exposure to extreme temperatures and hypoxia can protect neural operation against high temperature stress. They further suggest that the protective mechanism is a time-dependent process best revealed during prolonged exposure to extreme temperatures and is mediated by a neuromodulator such as serotonin.     

Electrical properties of developing oocytes of the migratory locust, Locusta migratoria.

The electrical properties of developing nonfertilized oocytes of Locusta migratoria were studied, using intracellular microelectrodes. The inseries potential of the combined oomembrane and of the follicular cells was about 20 mV in the youngest oocytes. It increased as the oocytes developed and it reached a plateau of about 50 mV before full maturation, generally four to seven oocytes away from the fully-developed terminal oocyte. Current-voltage relations were always linear for hyperpolarizing currents. Most oocytes exhibited, however, rectification to outward current. Input resistance values varied with oocyte size from about 5 X 10(6) ohm for young oocytes to about 0.2 X 10(6) ohm for the more developed ones. Some oocytes displayed a transient depolarization on turning off a hyperpolarizing step of current. This depolarization was not correlated with the size of the oocyte or with any observed morphological feature. Any two adjacent oocytes were electrotonically coupled. A single ovariole thus represented a longitudinal chain of developing oocytes which were connected electrically. This was supported by electron microscope observations which revealed junctions partially impermeable to lanthanum and gap junctions between the follicular cells themselves and between follicular cells and oocytes. The coupling coefficient was dependent on the direction of current flow. The attenuation of voltage along an ovariole was always greater at the distal than at the proximal side.     

The development of a collagenous connective tissue in the locust, Locusta migratoria

The ejaculatory duct of adult male locusts is surrounded by a thick layer of collagenous tissue. It starts to develop during the second instar when a group of cells come to lie adjacent to the basal regions of the epidermal cells which form the duct. From the fourth instar onwards, connective tissue is present between these cells. The matrix contains many collagen fibrils, some of which are very large and of irregular cross section. The banding periodicity is about 625 A. The cells differentiate into typical fibroblasts with highly dilated rough endoplasmic reticulum and small Golgi complexes. In the sexually mature adult, the cells are smaller and appear less active.