Fuels for flight in Locusta

In Locusta migratoria, carbohydrate is the predominant energy source for the first 20 to 30 min of flight. During this time carbohydrate is utilized at the rate of 120 μg/min, but thereafter it is used at only 8 to 9 per cent of the initial rate. Initially carbohydrate is not mobilized to any extent from body stores. Lipid, however, is mobilized (from stores in the fat body) in response to adipokinetic hormone released from the glandular lobes of the corpora cardiaca. The mobilized lipid is diglyceride, and 16 : 18 and 18 : 18 diglycerides predominate. The rate of diglyceride mobilization is rapid, both during flight and in response to extracts of corpora cardiaca. The release of adipokinetic hormone during flight has also been estimated and is found to be more rapid than previously suggested. It is calculated that the rate of lipid utilization increases during flight from virtually zero during the first few minutes to 35 μg/min (15 to 30 min) and finally reaches about 85 μg/min during the second 30 min of flight.     

Diacylglycerol-transporting lipoproteins and flight in Locusta

Evidence from chromatographic and heparin precipitation studies shows that the ‘heparin-soluble’ lipoprotein, A⁺, forms in the haemolymph during flight. In locusts flown continuously for 60 min, lipoprotein A⁺ occurs in the haemolymph at low concentrations but accumulates during a short rest period following flight. After injections of tissue extracts containing adipokinetic hormone (AKH), A⁺ accumulates in the haemolymph but disappears more rapidly in flying locusts than in resting locusts. This difference in the rate of disappearance of diacylglycerol from the lipoprotein A⁺ can be used to estimate its rate of utilization during sustained flight (approx. 100μg. min^?1 from 45–90 min of flight). It is suggested that lipoprotein A⁺ is the major carrier of diacylglycerol from the fat body to the flight muscles during prolonged flight. The steady state concentrations of total diacylglycerol and ‘heparin-soluble’ diacylglycerol during continuous flight are unaffected when tissue extracts containing AKH are injected before flight. This suggests that there is a close homeostatic control over the steady state concentration of haemolymph lipid during flight.     

Pre-imaginal flight motor pattern in Locusta

In a wind stream, larval stages of Locusta usually show a tonic muscle activity but they can also exhibit a rhythmic motor output. With ageing such a pattern can be released sooner, the trains become longer. The basic rhythm of 10 Hz does not change. The initial co-contraction of specific muscles is substituted later in development by an antagonistic recruitment. This activity resembles the flight motor pattern of young locusts which lack phasic sensory feedback from the wing region. Azadirachtin, an insect growth regulator, has been used to produce a permanent 5th larval instar. However, the extension of the last larval stage does not lead to a further development of the motor pattern to a level comparable to mature animals.     

Allatectomy and sexual receptivity in females of Schistocerca gregaria

The removal of the corpora allata from immature females of Schistocerca gregaria causes the insects to enter a phase of permanent defence towards courting males. Operated control females copulate often, although they do not appear to show an active form of sexual display or attraction. Implanted corpora allata fail to restore sexual receptivity or normal ovarial development in the allatectomized females due to the inactivity of the denervated glands. The observations are discussed in relation to findings in other locusts, grasshoppers, and crickets.     

Interrelationships between haemolymph lipid and carbohydrate during starvation in Locusta

During starvation in adult female Locusta, the haemolymph total lipid concentration increases markedly while that of the total carbohydrate decreases. The majority of the increased haemolymph lipid is diglyceride and 75% of this is associated with a high molecular weight lipoprotein (A⁺) which disappears rapidly after feeding when the total lipid concentration is restored to the normal resting value. The effect of feeding can be mimicked by injecting or feeding starved locusts with sugars but not with protein. The lowering of the haemolymph total diglyceride concentration in starved locusts by injection of carbohydrates is dose-related and, at doses in excess of 4 mg per locust, almost normal values (for fed locusts) are obtained within 6 hr. It is suggested that in the haemolymph there is an inverse relationship between the concentration of diglyceride and that of trehalose.     

The effects of allatectomy and ovariectomy on haemolymph proteins and lipid mobilization in Locusta

After ovariectomy the concentrations of diacylglycerol and protein in the haemolymph increase markedly. The increased diacylglycerol is associated with increased quantities of the ‘heparin-precipitable’ protein (lipoprotein A) that carries diacylglycerol in the blood of normal resting locusts. After the injection of adipokinetic hormone (AKH), the blood of ovariectomized locusts contains only slight quantities of the ‘heparin-soluble’ lipoprotein A⁺ whereas this forms in large amounts in the blood of sham-operated locusts after AKH injection. After allatectomy, the increase in the adipokinetic response is slower and the full level of responsiveness observed in sham-operated locusts is never attained. Nevertheless, allatectomized locusts develop a marked adipokinetic response which tends to stabilize as they age; it does not deteriorate as it does in aged sham-operated locusts.The effects of ovariectomy on blood metabolites can be prevented completely by allatectomy, but only partially by cautery of the cerebral neurosecretory cells. Treatment with a juvenile hormone analogue (JHA R-20458) counteracts the effects of allatectomy in ovariectomized locusts.     

The morphological and behavioural effects of precocene II on Locusta

The precocious adultiform larvae produced by topical applications of precocene II to early 4th instar Locusta were found to be intermediate between normal 5th instar larvae and normal adults on behavioural as well as morphological criteria. Precocious adultiforms walked significantly smaller distances in 2 hr than normal larvae, but slightly more than normal adults. The adultiforms jumped significantly greater distances without wing movements than normal larvae, but they also made frequent, short, adult-type jumps with wing movements. Most advanced adultiforms made jumps of both types whereas normal adults made only a small percentage of the larval-type jumps. Apparently the central nervous organisation of the adultiforms provides equally and separately for both larval and adult behaviour patterns, rather than compounding the two.Increasing doses of precocene above 100 μg produced more advanced adultiforms, and there was some indication that morphologically similar types produced by different doses may differ in their behaviour.     

Fluid and cation secretion by the Malpighian tubules of Locusta

In vitro preparations of Locusta Malpighian tubules are able to transport K⁺ against its concentration gradient. The ‘urine’ is slightly hyper-osmotic with respect to the bathing solution and the rate of secretion is inversely dependent on the osmotic pressure of the latter. The rate of fluid secretion increases with increasing temperature; being maximal at approx 40°C. The ionic composition of the secreted fluid, as indicated by Na⁺/K⁺ ratios, is altered by the presence of 1 mM ouabain in the bathing solution. Fluid secretion is inhibited by 1 mM ouabain. In addition, oxygen consumption by the Malpighian tubules is inhibited by either the presence of 1 mM ouabain or the absence of K⁺ in the bathing solution. The relationship between respiration, active transport and the Na⁺K⁺-activated ATPase is discussed.     

Lysozyme is stored in the granules of certain haemocyte types in Locusta

Lysozyme has been isolated from the serum of normal adult Locusta migratoria and antibodies were raised in rabbits against this molecule. Immunocytological techniques indicate that lysozyme is synthesized and stored in the two granular hemocyte types of this insect species: the typical granulocyte and the coagulocyte. The release of lysozyme appears to be associated with the process of coagulation.     

Quantitative studies on the activity of the corpora allata in adult male Locusta and Schistocerca

Juvenile hormone has been detected in the haemolymph and corpora allata of adult male Locusta and the haemolymph of adult male Schistocera by a modified Galleria bioassay. The hormone was readily detected in the haemolymph of insects immediately after the final ecdysis, but then became difficult to detect until 2 days prior to the onset of sexual maturation. In sexually mature insects the titre of juvenile hormone was maintained at a constant level. The corpora allata of adult male Locusta increased in size throughout adult life. The juvenile hormone content of the corpora allata was low during the period of somatic growth, but increased at the onset of sexual maturation. Sectioning of the nervi corporis allati I in insects immediately after the final ecdysis prevented the normal increase in size of the corpora allata, but did not render them inactive since juvenile hormone was detected in the haemolymph after the operation. The half life of juvenile hormone in the haemolymph of allatectomized adult male Locusta was 1 to 2 hr.     

The effects of precocene II on early adult development in male Locusta

Haemolymph protein synthesis and the accumulation of protein in haemolymph and accessory reproductive gland of normal adult male locusts were compared to that of males treated with precocene II. The precocene-treated insects showed a lower level of protein synthesis in both the fat body (source of haemolymph protein) and accessory reproductive gland. Topical application of juvenile hormone reversed the effects of precocene treatment. It would appear that the observed effects are the result of a reduction in juvenile hormone titre in the treated animals.     

Effect of the removal of the frontal ganglion on cellular structure in Locusta

An electron microscopical study has been made on the effects of frontal ganglionectomy on the structure of the cells of the fat body, midgut, and the neurosecretory cells of the pars intercerebralis in animals of the following treatments; operated, control operated, and starved. Both frontal ganglion removal and starvation produce changes in the fine structural appearance of the cells of the fat body and midgut. However, the changes are different in the two treatments; the mitochondris of the operated animals are swollen and the cisternae of the endoplasmic reticulum dilated. By contrast, the mitochondria are shrunken in starved animals and the endoplasmic reticulum sparse although numerous free ribosomes are present.     

Allatectomy and flight performance in maleLocusta migratoria

Summary Flight performance in locusts is a function of age, with its peak value at 18 days after emergence. While allatectomy retards the normal development of flight capability, it also has the effect of slowing down the decline in flight performance characteristic of operated control locusts as they age. Periodic topical application of synthetic juvenile hormone remedies the initial effect of allatectomy but its effectiveness wears off with age. The period of optimum flight performance is prolonged in locusts allatectomised when mature.A characteristic features of the flight pattern of immature-allatectomised and matureallatectomised locusts when flown about one week after the operation is a rapid decline in flight speed during the first 20 minutes of flight. Eventually, as the allatectomised locusts age, they assume the flight pattern of normal locusts and subsequent differences in flight performance between operated and normal locusts are confined to differences in flight intensity.Allatectomy has no marked effect on the preflight haemolymph total lipid and carbohydrate levels, the mobilisation of lipid and the amount of carbohydrate depleted. The quantity of lipid mobilised is, however, related to flight performance in both allatectomised and operated control locusts. Locusts which fly faster mobilise more lipid. The lipids mobilised by the adipokinetic hormone are 1618; 1818 and 1616 diglycerides in order of abundance. Allatectomy has no effect on the nature of these diglycerides released during flight.     

Qualitative and quantitative changes in Locusta haemolymph proteins and lipoproteins during ageing and adipokinetic hormone action

Changes in haemolymph proteins and lipoproteins during adipokinetic hormone action have been studied using polyacrylamide gel electrophoresis (PAGE) and a heparin/EDTA precipitation technique. During hormone action, the formation of A⁺ takes place at the expense of Ayellow and C_L-proteins, which decrease in free concentration in the haemolymph. Ayellow is heparin precipitable, whereas A⁺ precipitates with EDTA after prior treatment with heparin. After injection of adipokinetic hormone, heparin-precipitable protein (HPP) decreases after a delay of 10–15 min, but heparin/EDTA precipitable protein (HEPP) increases immediately. These changes occur in response to extracts of corpora cardiaca and to synthetic adipokinetic hormone, and are dose-dependent. Both the lipid and the C_L-protein content of the HEPP rise as its protein content increases. A⁺ formation does not occur in fifth-instar nymphs or newly emerged adults, but this response to adipokinetic hormone develops slowly as the adults mature.     

Effects of frontal ganglion removal and starvation on activity and distribution of six gut enzymes in Locusta

A study has been made on the effects of frontal ganglion removal and starvation on the activities and distribution of α- and β-glucosidase, α- and β-galactosidase, trehalase, and ‘trypsin’ in various regions of the alimentary canal of adult locusts. Both treatments resulted in a reduction in the amount of enzyme activity. In addition, the distribution of enzyme activity was changed by comparison with the operated control insects; the foregut of starved and operated animals showing a smaller proportion of the overall gut enzyme activity. The results are discussed in relation to the control of enzyme secretion.     

Ecdysteroids in adult males and females of Drosophila melanogaster

Ecdysteroid titres in whole flies and different tissues of adult male and female Drosophila were determined at various times after eclosion using a radioimmunoassay. The ecdysteroid titre decreased as the flies matured after eclosion. The differences in titre between males and females can be accounted for by their difference in body weight. The ecdysteroids were found to be distributed throughout several tissues. At eclosion not all of the ecdysteroid complement present could be accounted for by that found localised in tissues. After maturation of the flies the ecdysteroids in various tissues can account for the majority of that detected in whole-fly extracts. Ecdysteroids were produced during in vitro culture of various tissues, but the quantities detected were low by comparison with ring glands of wandering 3rd-instar larvae. Neither the ovaries nor the abdominal body walls (fat body) seem to be a major source of hormone, and they are only able to convert minute quantities of ecdysone to the biologically active form, 20-hydroxyecdysone, in vitro. The amounts of 20-hydroxyecdysone present were measured using high performance liquid chromatography and radioimmunoassay. We tentatively suggest that the differential experession of the yolk-protein-genes in the fat bodies of males and females does not result from differences in hormone titres between them.     

Chemical allatectomy of late Locusta embryos by a synthetic precocene and its effect on hopper morphogenesis

The anti-allatin substance, 7-ethoxy-precocene II (= “precocene III”) was topically applied to eggs of Locusta migratoria migratorioides with fully grown embryos in stage XX (about 64 ± 4% of the whole period of egg development). A day after precocene application the eggs were washed for 10–15 s in acetone and then transferred to clean containers for removing precocene residues and for preventing contamination at hatching. The treatment induced prothetelic morphogenetic disturbances which became overt in the subsequent hoppers; the effect was dose dependent and the ED₅₀ (= effective anti-allatin dose 50%) was low, 20.5 μg precocene III per g fresh egg weight (= 0.37 μg per egg). Quite similar results were obtained following application of precocene III to eggs with embryos in stage XXI (73 ± 4% of the egg development). These findings and direct examination of histological sections of the embryonic corpora allata demonstrated that precocene chemically allatectomizes late Locusta embryos. The lethal effect of precocene III was dependent on the washing. When the eggs were washed in acetone a day after application, mortality did not occur in a dose-dependent way; even the highest dose applied, 256 μg precocene III per egg (= 14405 μg per g fresh weight), was less than the LD₅₀ (lethal dose 50%). In contrast, without washing mortality was dose dependent, but it occurred later, at or after hatching; the LD₅₀ was 1334.9 μg per g (= 22.7 μg/egg). The results show that the late embryos are highly susceptible to the anti-allatin effect of the precocene, but are extremely insusceptible to its lethal effect; toward hatching, however, susceptibility to the lethal effect becomes marked.With doses between 45–14405 μg precocene III per g fresh egg weight, the anti-allatin effect became overt by a quite-uniform belated morphogenetic response. All hoppers which hatched from precocenetreated eggs were morphogenetically normal in the 1st instar and in the beginning of the 2nd instar, but the duration of the 2nd instar was almost doubled and at the end of this instar over 96% of the locusts died in the moult, being unable to shed the exuvia. Artificial removal of the apolyzed old cuticle revealed 3rd instar prothetelic adultiforms. These results and some data in the literature indicate that allatectomy of the embryo does not result in prothetelic morphogenetic disturbances in the 1st and early 2nd instar larvae and may impose the question what is the role of the juvenile hormone in late embryos and early larvae.     

Sexual selection and reproductive success in males of the bicolor damselfish, Eupomacentrus partitus (Pisces: Pomacentridae)

Relative reproductive success of male bicolor damselfish, Eupomacentrus partitus, was measured as the number of egg batches obtained per week per male during a 14-month field study. Large and consistent differences in spawning success, observed in seven colonies containing 23 males, demonstrated that mating was non-random. Various physical and behavioural characteristics of males were also monitored. Significant correlations (P<0.05) were found between reproductive success and four variables: frequency of courtship, inter- and intraspecific aggression, and total length. The importance of other factors such as size and location of male territories and time allotment to various activities was also considered. The interactions between these variables, as well as preliminary observations of spawning, suggest that intersexual selection, that is female choice of mates, is the dominant process in sexual selection in this species.     

Courtship processing in Drosophila melanogaster. II. An adaptation to selection for receptivity to wingless males

Measurements of the locomotor activity in courtship cells of female types from the experiment reported in the previous paper disclosed that the presence of a second animal reduced the activity level of the female being measured. The reduction in activity was greater when the second animal was a courting wingless male than when it was a female. Such lowering of the female's activity during courtship was found to be related to subsequent receptivity to wingless males. Strain differences in activity score and rate of decline in activity over the test period confirmed that this system was probably responsible for the selection gain observed. Further experiments attempted to clarify the causal factors associated with these phenomena.