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Neuropeptides, amines and amino acids in an elementary insect ganglion: functional and chemical anatomy of the unfused abdominal ganglion.

The insect ventral nerve cord consists of metamerically repeated ganglia subserving the thoracic and abdominal segments. The abdominal ganglia control basic functions such as respiration, circulation, heartbeat, diuresis, hindgut motility, functions of the genitalia and ovipositor and abdominal posture. Some of this control is by efferent innervation of target tissues but hormonal control also is exerted by abdominal neurosecretory cells via release from neurohemal organs or other release sites. The present review summarizes what is known about the distribution of neurotransmitters, monoamines and neuropeptides in the abdominal ganglia of different insect species. Special emphasis is on the unfused abdominal ganglion, since this is the least complex of all central ganglia and therefore may reveal the minimum number of neuroactive compounds utilized in neurotransmission, neuromodulation and neurohormonal control. Both GABA and glutamate are present in both interneurons and motoneurons, whereas biogenic amines such as serotonin, dopamine and histamine are found primarily in interneurons (although some cases of sensory cells and efferent neurons are known). Octopamine can be seen both in interneurons, efferent neurons and neurosecretory cells. A large number (about 20 different main types) of neuropeptides has been indicated in abdominal ganglia. Each peptide has a very specific distribution pattern. Depending on the peptide type, the localization is known to be in interneurons, neurosecretory cells or motoneurons, or combinations of these. The structure and known functions of the different neuropeptides in different insect species are summarized in some detail. Both GABA and glutamate appear to have roles as fast neurotransmitters, whereas amines and neuropeptides seem to have modulatory roles both within the CNS and at peripheral targets. After a comprehensive overview of different substances in studied insect species, the unfused abdominal ganglia from the moth Manduca sexta, locusts and cockroaches are dealt with in some detail and a comparison is made with insects possessing fused abdominal ganglia such as blowflies and Drosophila. Some emphasis is made of the presence of neuroactive compounds in neurosecretory cells and other identifiable neurons for which physiological analysis is feasible.

Amino Acids↗

Glucose metabolism in an insect Manduca sexta and effects of parasitism.

The metabolism of [1-13C]glucose was examined during the last larval stadium of an insect Manduca sexta (Lepidoptera: Sphingidae) parasitized by Cotesia congregata (Hymenoptera: Braconidae). Following injection, the isotopically substituted glucose was metabolized at a significantly lower rate by parasitized larvae than by normal, control insects. 13C enrichment was principally observed in [1-13C]trehalose in both groups. Randomization of the label at the triose phosphate step was evidenced by incorporation of 13C into C6 of trehalose. Parasitized and control larvae both synthesized [1,6-13C]glycogen but the relative amount of label observed in parasitized larvae was greater. The ratio of C6/C1 enrichment in trehalose and glycogen was significantly less in parasitized larvae. The rate of labelled trehalose and glycogen synthesis was relatively high when compared with the estimated rate of glycolytic glucose oxidation, and the difference in C6/C1 enrichment ratio between normal and parasitized insects was, therefore, not reflective of a difference in the rate of substrate cycling, but rather, was due to the increased synthesis of [1-13C]glycogen in parasitized larvae when compared with controls. Inhibition of glycolysis by administration of iodoacetate to normal larvae resulted in an increase in the incorporation of 13C into glycogen relative to glucose metabolized, suggesting that inhibition of glycolysis may be responsible for the higher level of glycogen synthesis observed in parasitized insects. In control larvae, significant 13C enrichment from [1-13C]glucose was observed in fat, but no evidence of lipogenesis was observed in parasitized insects. Iodoacetate had no observable effects on the relative amount of 13C incorporated into fat. Malonic acid and cyanide resulted in accumulation of 13C from [1-13C]glucose in several TCA cycle intermediates of normal larvae, but had little effect on the relative enrichments of trehalose, glycogen and fat.

Animals↗

A comparative catabolism study of isoleucine by insect and mammalian tissues.

The metabolism of [U-14C]isoleucine was examined in different tissues of five species of lepidopteran and four species of non-lepidopteran insects. Slices of fat body, epidermis, Malpighian tubule, gut, and muscle were incubated in a culture medium containing [U-14C]isoleucine; the medium was analyzed by ion-exclusion LC to quantify labeled metabolites. Tissues of lepidopteran insects secrete high levels of metabolites including 2-keto-4-methylvalerate, 2-methylbutyrate, propionate, and acetate. Tissues of non-lepidopteran insects secrete low amounts of these acids. Analysis of isoleucine transaminase activity in selected tissues of non-lepidopteran insects indicated that those tested contain significant activity. These results demonstrate that tissues of lepidopteran insects have a unique ability to secrete short chain acids, derived from isoleucine, into the medium. The secretion of propionate correlates with the ability to synthesize ethyl-branched juvenile hormones and indicates the presence of an efficient transport system for short chain acids. We also monitored the secretion of acidic metabolites of isoleucine by different tissues of the rat. Muscle was most active in secreting keto acid whereas heart secreted high levels of 2-methylbutyrate. Negligible quantities of metabolites of isoleucine were secreted by the liver.

Animals↗

Cuticle degrading proteases from insect moulting fluid and culture filtrates of entomopathogenic fungi.

Insects degrade their own cuticle during moulting, a process which is catalysed by a complex mixture of enzymes. Entomopathogenic fungi infect the insect host by penetration of the cuticle, utilizing enzymatic and/or physical mechanisms. Protein is a major component of insect cuticle and a major recyclable resource for the insect and, therefore, represents a significant barrier to the invading fungus. To this end, both insects and entomopathogenic fungi produce a variety of cuticle degrading proteases. The aim of this paper is to review these proteases and to highlight their similarities, with particular reference to the tobacco hornworm, Manduca sexta, and the entomopathogenic fungus, Metarhizium anisopliae.

Animals↗

Use of cowpea trypsin inhibitor (CpTI) to protect plants against insect predation.

Transgenic tobacco plants expressing a cowpea trypsin inhibitor gene have enhanced levels of insect resistance to a variety of insect pests. Furthermore, insect bioassay has shown the cowpea trypsin inhibitor to have anti-metabolic activity to insect pests of the orders Lepidoptera, Coleoptera and Orthoptera. The advantages and disadvantages of this approach to developing insect resistant crops is discussed in relationship to other methods, including conventional plant breeding and chemical control. Eventually it is hoped that African farmers will benefit from this industrially sponsored research.

Journal Article↗

Insect control with baculoviruses.

Baculoviruses have been researched extensively for insect control. Three of their features have been particularly attractive: their host specificity and consequential environmental safety, their virulence in host insects, and their capability for causing disease epizootics. There have been four approaches to their use for insect control: as microbial insecticides for short-term insect population suppression, through seasonal colonization or a recurring "booster shot" for control of more than one pest generation, through introduction-establishment where the viral species or strain was not indigenous, and through environmental manipulation to make the ecosystem more favorable for viral epizootics. Actual usage of baculoviruses in pest management has been disappointing, particularly with the microbial insecticide approach, primarily for three reasons: economics, slow speed of kill, and adverse effects of the environment on the viruses. The recombinant-DNA revolution has greatly increased the prospects for baculoviruses in insect pest management.

Journal Article↗

Monoamine uptake in insect synaptosomal preparations.

Biochemical studies of mammalian synaptosomal nerve fractions indicate the existence of multiple transporter proteins important for the termination of synaptic transmission by each of several monoamines. In insects, however, data on monoamine uptake has been limited to the study of whole tissue preparations, making it unclear whether neuronal (as opposed to glial) uptake is a significant mechanism in the insect. The present experiments elucidate the difficulties that have limited the use of insect synaptosomal preparations for characterizing amine reuptake. Key procedural improvements, including the utilization of carrier protein for tracer separation and the use of receptor antagonists to decrease non-specific membrane binding are described. With these and other modifications, reproducible sodium-dependent and cocaine-inhibitable dopamine and octopamine uptake are described in synaptosomal-containing preparations from insect brain and ganglia. These studies therefore support the existence of specific Na(+)-dependent uptake mechanisms in insect neurons.

Animals↗

What can we learn from prostaglandins and related eicosanoids in insects?

Eicosanoids are oxygenated metabolites of three C20 polyunsaturated fatty acids (20:3n-6, 20:4n-6, and 20:5n-3). While eicosanoids are very well known in mammalian systems, mostly due to their pharmaceutical interest, there is increasing recognition of the significance of these compounds in insects and other invertebrates. In this paper we consider four major concepts emerging from work on eicosanoids in invertebrates. First, the biological significance of eicosanoids extends far beyond their physiological and pathophysiological actions in human and veterinary medicine. Second, we can greatly improve our understanding of eicosanoids in insects by integrating our work on insects into ongoing studies of other invertebrates. Third, some eicosanoid actions may be fundamental to animals. Fourth, the biochemistry of eicosanoids in insects and other invertebrates can differ from expectations based on the mammalian background. Finally, we point to an uncharted frontier in insect studies--the biochemical mechanisms of eicosanoid action--by drawing attention to some of the work on eicosanoid receptors in mammalian systems.

Animals↗

An experiment using neutron activation analysis and a rare earth element to mark cotton plants and two insects that feed on them.

Studies on insect dispersal and other behaviors can benefit from using markers that will not alter flight and fitness. Rare earth elements, such as samarium (Sm), have been used as ingested markers of some insects and detected using neutron activation analysis (NAA). In this study, samarium nitrate hexahydrate was mixed into artificial diet for boll weevils, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), at different dosages and in water used to irrigate cotton, Gossypium hirsutum L. Samarium was detected in adult boll weevils fed on the samarium-labeled diet, but not after 5 or 10 days of being switched to non-labeled diet, even if the insects were given labeled diet for as long as 7 consecutive days. Introduced in irrigation water, 1% samarium (m/m) was detectable in cotton squares and leaf tissue. However, boll weevil adults fed samarium-labeled squares did not retain detectable levels of samarium, nor did boll weevil adults reared to adulthood from samarium-labeled squares. Fourth instar beet armyworms, Spodoptera exigua (Hübner) (Noctuidae: Lepidoptera), fed on samarium-labeled cotton leaves obtained enough samarium for NAA detection, but adult moths reared from them did not have detectable amounts of samarium. Although samarium can be useful as a marker when insects are presented with a continuous pulse of the label, elements that are assimilated by the insect would be more useful if a continuous infusion of the marker cannot be provided.

Animals↗

Sperm viability matters in insect sperm competition.

Experimental studies in insects have shown how sperm competition can be a potent selective force acting on an array of male reproductive traits . However, the role of sperm quality in determining paternity in insects has been neglected, despite the fact that sperm quality has been shown to influence the outcome of sperm competition in vertebrates . A recent comparative analysis found that males of polyandrous insect species show a higher proportion of live sperm in their stores . Here, we test the hypothesis that sperm viability influences paternity at the within-species level. We use the cricket Teleogryllus oceanicus to conduct sperm competition trials involving prescreened males that differ in the viability of their sperm. We find that paternity success is determined by the proportion of live sperm in a male's ejaculate. Furthermore, we were able to predict the paternity patterns observed on the basis of the males' relative representation of viable sperm in the female's sperm-storage organ. Our findings provide the first experimental evidence for the theory that sperm competition selects for higher sperm quality in insects. Between-male variation in sperm quality needs to be considered in theoretical and experimental studies of insect sperm competition.

Animals↗

Functional conservation of an insect odorant receptor gene across 250 million years of evolution.

Pest insects have a profound negative impact on agriculture and human health. Significant global losses of crops, stored agricultural products, timber and livestock can be attributed to damage and destruction by insects . Blood-feeding insects such as mosquitoes, flies and ticks transmit many of humanity's most devastating infectious diseases. Insect-borne diseases account for more than one million annual fatalities, and insect-associated illnesses surpass 300 million annual reported cases . The medical and economic impact of these animals can be ascribed in part to the sensitivity and selectivity of their olfactory systems, essential for location of their preferred plant and animal hosts.

Amino Acid Sequence↗

Nutrition influences growth and virulence of the insect-pathogenic fungus Metarhizium anisopliae.

Nutrition influenced growth, sporulation and virulence of the insect pathogenic fungus, Metarhizium anisopliae. Virulent conidia were produced on susceptible insect hosts, 1% yeast extract, 2% peptone, osmotic stress medium (OSM) and CN 10:1 medium. Several strain independent markers were identified that could be used to predict the virulence of M. anisopliae conidia. Virulent conidia typically had high levels of spore bound Pr1, an important cuticle degrading protease, and high germination rates. We also show for the first time that virulent conidia have an endogenous CN ratio below 5.2:1. Real Time PCR revealed that virulent conidia from insects contained significantly higher levels of transcripts of pr1 A and other pathogenicity-related genes than inoculum from artificial media. Of the artificial media studied, 1% yeast extract medium yielded the most virulent conidia, these had higher levels of transcripts of these pathogenicity-related genes than the least virulent conidia from the high conidia yielding CN 35:1 medium (=SDA), however, the levels were significantly lower than those in insect-derived conidia. Our study shows for the first time that the passaged inoculum is virulent irrespective of the original culture medium or insect host. Virulent conidia were consistently produced on OSM even though growth and sporulation were poor. We postulate that starvation conditions, whether in vivo or in vitro, results in de-repression of Pr1 and that elevated levels of this enzyme enhance fungal virulence.

Animals↗

Lipid uptake by insect oocytes.

Approximately 30-40% of the dry weight of an insect egg consists of lipid, mostly triacylglycerol (TAG). Although this lipid is essential for the energy needed by the developing embryo, little is known about the mechanism that leads to the accumulation of TAG in the insect egg. Insect oocytes can readily synthesize TAG from free fatty acids (FFAs) and glycerol, however, de novo synthesis of FAs by the oocyte is marginal. Hence, FAs have to be imported from the fat body or the diet. Insect hemolymph contains two lipoproteins that transport lipids, lipophorin and vitellogenin. Both are taken up via endocytosis by the oocyte, however, this provides only about 10% of the egg's lipid reserves. The rest is unloaded from circulating lipoprotein particles at the oocyte surface in the form of diacylglycerol (DAG), the major lipid transport form in insects, or as FFA. The mechanism of lipoprotein unloading at the oocyte surface is currently unclear. Possible roles of the lipid transfer particle (LTP), FA transporters, and lipoprotein lipase activity are discussed.

Animals↗

Insect haemocytes: what type of cell is that?

Classification of insect larvae circulating haemocytes is the subject of controversy, and the terminology used to designate each cellular type is often different from one species to another. However, a survey of the literature on insect haemocytes suggests that there are resemblances for most of the cell types and functions, in different insect species. In this review paper, we compare the structure and functions of circulating haemocytes in those insect species that are, by far, the most often used species for insect physiology studies, i.e. lepidopteran species and Drosophila. We show that there is high degree of homology of haemocyte types and suggest possible synonymies in terminology among species from these taxa.

Animals↗

Variations in UV-B tolerance and germination speed of Metarhizium anisopliae conidia produced on insects and artificial substrates.

Solar ultraviolet radiation (UV-A and UV-B) is a major factor in failure of programs using the insect pathogenic fungus Metarhizium anisopliae as a biological control agent. Studies were conducted to determine if growth conditions, viz. artificial (agar media or rice grain) or natural (infected insects) substrates for conidial production affect two traits that directly influence performance of conidia after field application: tolerance to UV-B radiation and conidial germination speed. Conidia of two isolates (ARSEF 23 and ARSEF 2575) of M. anisopliae var. anisopliae produced on potato dextrose agar plus yeast extract (PDAY) or on fungus-killed larvae of two insect species, Galleria mellonella and Zophobas morio, were inactivated by exposure to UV-B radiation. Conidia of both isolates when produced on insect cadavers were significantly more sensitive to UV-B radiation than conidia produced on PDAY. Also, conidia from insect cadavers germinated slower than those from PDAY cultures. A comparison of conidia from artificial substrates showed that conidia produced on Czapek's and Emerson's YpSs agar media or rice grains had higher tolerance to UV-B radiation and germinated faster than conidia raised on PDA and PDAY. Accordingly, the growth substrate and nutritional environment in which conidia are produced influences M. anisopliae conidial UV-B tolerance and speed of germination; and manipulation of these variables could be used to obtain conidia with increased tolerance to UV-B radiation and shorter germination times.

Animals↗

Insect-parasitic nematodes: from lab curiosities to model organisms.

Interest in studying insect-parasitic nematodes was originally focused on their potential as biological control agents of insects and other arthropod pests. Now, after 30 years of intense basic and applied research, realization of the practical use of insect-parasitic nematodes, particularly of entomopathogenic nematodes and their symbiotic bacteria, has spurred developments across a far broader scientific front. We are now entering a new era of discovery in which tools of molecular genetics are being increasingly used to address a range of biological questions. The knowledge gained from these efforts will directly benefit the practical application of insect-parasitic nematodes as more effective biopesticides. Moreover, these studies will advance these nematodes as unique and intrinsically interesting biological model systems not only for basic research but also in applied fields such as plant health, human medicine, pharmaceutical bioprospecting, and genetic engineering. In this review, the past and current state of insect-parasitic nematode research is summarized. Future research priorities and goals are identified and discussed.

Animals↗

Use of light, scanning electron microscopy and bioassays to evaluate parasitism by entomopathogenic fungi of the red scale insect of palms (Phoenicococcus marlatti Ckll., 1899).

We have evaluated the parasitism of the red scale insect of the date palm (Phoenicococcus marlatti) by entomopathogenic fungi, using light microscopy (LM), scanning electron microscopy (SEM) and low temperature scanning electron microscopy (LTSEM). Beauveria bassiana, Lecanicillium dimorphum and Lecanicillium cf. psalliotae, were inoculated directly on the scale insects or on insect infested plant material. We found that L. dimorphum and L. cf. psalliotae developed on plant material and on scale insects, making infection structures. B. bassiana was a bad colonizer of date palm leaves (Phoenix dactylifera L.) and did not parasite the scale insects.

Cocos↗

Time reduction and process optimization of the baculovirus expression system for more efficient recombinant protein production in insect cells.

Rapid expression of recombinant proteins for structure determination is one of the major challenges in pharmaceutical and academic research, since the number of potential drug targets has increased significantly in the last decade. Despite the fact that the baculovirus expression vector system is widely used for this purpose, the system is hampered by three very slow and tedious procedures, namely generation of high titer baculovirus stock, determination of the virus titer and discovery of the best conditions for protein expression. We herein describe the development of the ultraBac system to address and overcome these issues for protein expression in insect cells. We have established a new baculovirus expression technology for insect cells that is based on co-expression of GFP with target genes, a new regime for cell culturing and a highly efficient purification and enrichment procedure for recombinant baculovirus particles. Co-expression of GFP is used to monitor the infection of insect cells, to simplify titer determination and to optimize expression conditions. The new regime for cell culturing with increased viability of non-infected insect cells and its combination with the massive enrichment of virus particles via high-speed centrifugation enables the production of large amounts of recombinant virus in a very short period of time. By combining these techniques and by using the bicistronic vector pUltraBac-1, we have been able to cut the time-lines for protein expression in insect cells by half, approaching those for protein production in Escherichia coli. This new expression system is a significant step forward towards industrialized protein production in both, industry and academia.

Animals↗