W.J.A. Van Marrewijk

Insect flight muscle metabolism

The flight of an insect is of a very complicated and extremely energy-demanding nature. Wingbeat frequency may differ between various species but values up to 1000 Hz have been measured. Consequently metabolic activity may be very high during flight and the transition from rest to flight is accompanied by an increase of 50-100-fold in metabolic rate. Small mammals running at maximal speed and flying birds achieve metabolic rates exceeding resting levels by only 7-14-fold. The exaggerated metabolic rate during insect flight is not accompanied by an oxygen debt, which implies -apart from metabolic adaptations- ample availability of oxygen in the organs responsible for flight. Metabolic rate therefore can be estimated, apart from the depletion of fuel depots, by rates of oxygen consumption.

Role of Lipids in Energy Metabolism

Most reviews of lipid metabolism in insects have covered all classes of lipid compounds. Since lipids are generally defined as substances poorly soluble in water but soluble in organic solvents, the authors had to deal with compounds with divergent physiological functions, e.g., phospholipids and pheromones. The subject of this chapter limits the lipoidal substances to be discussed to those that provide a direct source of metabolic energy, though we recognize that other important lipid classes, for example those contributing to (sub)cellular components, are also essential for metabolism.

Regulation of glycogenolysis in the locust fat body during flight

Glycogen reserves in the fat body of Locusta migratoria decrease dramatically during the first two hours of flight. In fat body of rested locusts only 10% of glycogen phosphorylase occurs in the active form. The enzyme is activated significantly during flight, when up to one-third of the total phosphorylase becomes active. Phosphorylase activation can also be accomplished by injection of corpus cardiacum extracts, to give a maximum of circa 75% active enzyme. Locust fat body is shown to contain protein kinase activity, which can be activated both by cyclic AMP and cyclic GMP. Apparent Ka-values are 0.13μM for cyclic AMP and 0.16 μM for cyclic GMP. Results are discussed in relation to regulation of substrate utilization during flight.

A dual function of alcohol dehydrogenase in Drosophila

Alcohol dehydrogenase (ADH) of Drosophila not only catalyzes the oxidation of ethanol to acetaldehyde, but additionally catalyzes the conversion of this highly toxic product into acetate. This mechanism is demonstrated by using three different methods. After electrophoresis the oxidation of acetaldehyde is shown in an NAD-dependent reaction revealing bands coinciding with the bands likewise produced by a conventional ADH staining procedure. In spectrophotometric measurements acetaldehyde is oxidized in an NAD-dependent reaction. This activity is effectively inhibited by pyrazole, as specific inhibitor of ADH. By means of gas chromatographic analysis a quick generation of acetate from ethanol could be demonstrated. Our conclusion is further supported by experimental results obtained with either purified ADHF enzyme or genotypes with or without ADH, aldehyde-oxidase, pyridoxal-oxidase and xanthine-dehydrogenase activity. These results are discussed in relation to ethanol tolerance in the living organism in particular with respect to differences found between ADH in Drosophila melanogaster and D. simulans, and in relation to the possible implications for the selective forces acting on ADH-polymorphism.

Insect adipokinetic hormones

Peptides with adipokinetic (and usually carbohydrate-mobilizing) potency have been demonstrated in various insects, including Locusta migratoria, Schistocerca gregaria, Manduca sexta, Danaus plexippus and Periplaneta americana. As far as characterized by now the adipokinetic factors are blocked peptides, consisting of eight to ten amino acid residues. In locusts the adipokinetic hormones are synthesized in the glandular lobe of the corpus cardiacum and released into the haemolymph in response to flight stimuli. This release is under direct control of neurons, the cell bodies of which are located in the lateral areas of the protocerebrum, while their axons run via the nervi corporis cardiaci II into the glandular lobe. Hormone release is modulated by axons present in the nervi corporis cardiaci I as well as by the haemolymph trehalose concentration. Trehalose apparently exerts its influence via a neuronal network present in the corpus cardiacum. The fat body is the main target organ of the adipokinetic hormones, which are involved in both mobilization and release of flight substrates from fat body stores, i.e., trehalose from glycogen and diacylglycerol from triacylglycerol. Lipid release is accompanied by haemolymph lipoprotein conversions.

Regulation of glycogen phosphorylase activity in fat body of Locusta migratoria and Periplaneta americana

Saline extracts of the corpus cardiacum (CC) of Locusta migratoria activate glycogen phosphorylase in locust fat body. The response of phosphorylase to CC extracts and to synthetic adipokinetic hormone (AKH) suggests that the factor responsible for the activating effect of the CC on phosphorylase is AKH, supplemented to a minor degree with Compound II. Octopamine does not influence fat body phosphorylase activity in locusts, however, it elicits a rapid short-term hyperlipemia. In cockroaches, Periplaneta americana, injection of octopamine results in a strong activation of fat body phosphorylase within 1 min. Cockroach CC extract exerts a more prolonged effect on phosphorylase activity than does octopamine.

FREE AMINO ACIDS IN THE SHRIMP CRANGON CRANGON AND THEIR OSMOREGULATORY SIGNIFICANCE

Abstract Measurements of the concentrations of ninhydrin positive substance ( nps ) and of the individual free amino acids in muscle and haemolymph of Crangon crangon , adapted to various salinities at different temperatures, are recorded, and their significance as osmotic effectors is evaluated. The pattern of nps regulation in muscle under various environmental conditions being strictly analogous with that of the haemolymph osmotic regulation, illustrates its importance to the osmotic adjustment of the cells in response to changes in environmental conditions. The salinity-induced variation in nps in turn reflects the total concentrations of the 3 most abundant amino acids: glycine, proline and alanine. Parallel measurements of muscle and haemolymph nps during the course of adaptation to hyper- and hypo-osmotic salinities evidence an ecologically-significant rapid response of the isosmotic intracellular regulation, and indicate an exchange of nitrogenous material across the cell membranes. The results are discussed comparatively and with regard to the theories on the regulation of the nitrogenous contents and osmotic concentrations in the cells.

Hypertrehalosaemic and hyperlipaemic responses to adipokinetic hormone in fifth larval instar locusts, Locusta migratoria

In fifth instar larvae of Locusta migratoria the haemolymph lipid concentration is elevated after injection of adipokinetic hormone (AKH). This hyperlipaemic response in larvae remains substantially lower than in adults; over 75% of the mobilized lipid consists of diacylglycerol. In addition, unlike adult locusts, fifth instar larvae also exhibit a consistent, though moderate, hypertrehalosaemic response to AKH. The increases of both lipid and carbohydrate concentrations in larvae are dose-dependent, showing a significant linear regression on log dose in the range 0.2–20 pmol AKH. Glycogen phosphorylase in the fat body of fifth instar larvae as well as young adults is activated on injection of AKH, the percentage active phosphorylase increasing linearly with log dose in the range 0.04–20 pmol AKH. For a given response, a somewhat higher dose of AKH is needed in larvae than in young adults. Fat body glycogen phosphorylase is strongly activated during the period of the larval-adult ecdysis, when active phosphorylase accounts for almost half of the total enzyme, which is approximately ten times more than it is two days before, and two days after the ecdysis. The corpora cardiaca of fifth larval instar locusts already possess the potencies to elevate carbohydrate and lipid concentrations in larval haemolymph, and to activate fat body glycogen phosphorylase.

Adipokinetic hormone is dependent on extracellular Ca2+ for its stimulatory action on the glycogenolytic pathway in locust fat body in vitro

Abstract Inclusion of glucose or trehalose in the medium during the incubation of locust fat body in vitro leads to a reduction of the relative amount of active (AMP-independent) glycogen phosphorylase. The presence of adipokinetic hormone (AKH I) results in a rapid activation of phosphorylase, reaching a maximum within 5 min. This AKH effect is highly dependent on added Ca 2+ , and requires ⩾ 1 mM Ca 2+ for maximal enzyme activation. Ca 2+ alone has no effect on phosphorylase activity, but it does activate the enzyme when the ionophore A23187 is also included in the medium. In a cell-free system from locust fat body the activation of endogenous phosphorylase by phosphorylase kinase is stimulated by Ca 2+ . Activity of the latter enzyme can be increased further by high doses of calmodulin. Both in the presence and in the absence of external calmodulin, the calmodulin antagonist trifluoperazine has an inhibitory effect on phosphorylase kinase. Results are discussed in relation to the possible mechanisms underlying hormonal control of glycogenolysis.

signal transduction of adipokinetic hormones involves Ca2+ fluxes and depends on extracellular Ca2+ to potentiate cAMP-induced activation of glycogen phosphorylase

Adipokinetic hormone (AKH)-induced mobilization of insect fat body glycogen occurs through activation of glycogen phosphorylase. In the migratory locust, signal transduction of AKH-I, -II and -III has been shown to involve the formation of cAMP. In the present study, we show that both the elevation of fat body cAMP levels and the activation of phosphorylase by the three AKHs in vitro depend on the presence of extracellular Ca2+; in the absence of Ca2+ in the medium, no phosphorylase activation occurs, whereas a concentration of at least 1.5 mM Ca2+ in the medium is required for maximal activation by each of the hormones. Furthermore, we show that AKH-I, -II and -III increase the influx of extracellular calcium into the fat body, as well as the efflux of cytosolic calcium from the fat body into the medium within 1 min of incubation. Although the time courses of their effects and the maximal responses to massive doses (40 nM) of the three hormones do not differ, AKH-III induces the highest increase in Ca2+ efflux when applied in a physiological dose (4 nM). No difference in the levels of Ca2+ influx induced by 4 nM of the hormones was observed. Quantitative analysis of the data suggests that the AKH-induced influx is larger than the efflux, implying a net rise in the fat body Ca2+ concentration.