The alteration of juvenile hormone titre inSpodoptera litura (F.) due to a baculovirus infection

Summary The haemolymph juvenile hormone levels ofSpodoptera litura were remarkably low (540 Galleria units (GU)/ml) at the last larval moult as well as prior to pupation (194 GU/ml). During the last intermoult period this was 2600 GU/ml for a 24 h-period. On the other hand, the JH level in the haemolymph of NPV-infected last instar larvae was initially 1740 GU/ml but was maintained at 2400–2600 GU/ml during the next 48 h. Finally, the JH titre fell to 1393 GU/ml, but only prior to death. The failure of the diseased larvae to undergo the larval pupal moult is ascribed to the alteration of the JH titre in the haemolymph.     

Juvenile hormone titers in penultimate and last instar larvae ofPieris brassicae andBarathra brassicae,in relation to the effect of juvenoid application

Summary JH titer was determined in the haemolymph of penultimate and last instar larvae ofPieris brassicae L. andBarathra brassicae L. The differences we observed were consistent with physiological differences between the two species.The authors feel indebted to Prof.V. J. Brookes for useful comments.     

Lipid transport in the migrating Monarch butterfly,Danaus p. plexippus

Summary The transport of lipid in the haemolymph of the Monarch butterfly during its fall migration was examined. Diglyceride was the major lipid class of 2 electrophoretically distinct lipoprotein fractions in both males and females. Triglycerides, hydrocarbons, free fatty acids, phosphatidyl cholines and phosphatidyl ethanolamines were minor components of these lipoproteins. Differences in lipid transport attributable to sex were not detected.This study is a contribution of the Missouri Agricultural Experiment Station, Journal series No. 8520.     

Juvenile hormone catabolism inManduca sexta: Homologue selectivity of catabolism and identification of a diol-phosphate conjugate as a major end product

We studied time-dependent metabolism of (10R)-[³H] juvenile hormone (JH) III and (10R, 11S)-[³H]JH I injected intoManduca sexta larvae; the hormones are metabolized to polar metabolites, expecially the JH acid-diol, and an unknown. Products were analyzed using a reversed-phase liquid chromatography assay. (10R)-JH III is metabolized much more rapidly than (10R, 11S)-[³H]JH I, whether injected seperately or as a mixture of hormones. The unknown metabolites of JH I and JH III were identified as phosphate conjugates of JH I and JH III diol by tandem mass spectral analysis of isolated samples. The phosphate conjugate of JH I diol is the principle end product of JH I metabolism.     

The influence of juvenile hormone on the feeding behaviour of last instar larvae of the codling moth,Laspeyresia pomonella (Lep., Tortricidae), reared under different photoperiods

Summary Duration of the feeding stage and corresponding weight increase during the last larval instar of the codling moth,Laspeyresia pomonella, are controlled by JH. Larvae reared under short day conditions have a relatively high titer of JH during the last larval instar and enter diapause as mature larvae. They feed longer and become heavier than larvae reared under long day conditions, which have no JH during the last larval instar and pupate when mature. By application of the JH mimetic Altosid^® during the first 2 or 3 days of the last larval instar, the duration of feeding activity of larvae reared under respectively long and short day conditions was prolongated and the larvae became significantly heavier. The feeding behaviour could only be influenced by the juvenoid as long as the feeding activity of the larvae had not yet ceased.     

The transfer of juvenile hormone from male to female during mating in the Cecropia silkmoth

Summary The juvenile hormone (JH) stored in the accessory sex glands (ASG) of adult maleHyalophora cecropia (L.) originates both from sequestration of circulating hormone and from JH synthesized de novo in the ASG from JH acid taken up from the hemolymph. The secretions present in the lumina of the ASG contain most of the accumulated JH. During mating, endogenous JH, labeled biosynthetically via injected [³H-methyl]-methionine, is transferred along with the other seminal material to the bursa copulatrix of the female. The physiological significance of the JH transfer remains unknown.Research was supported by a grant from the National Science Foundation (PCM72-01892) and Organized Research Funds from Texas A&M University.This work was done as partial fulfillment for the degree of Doctor of Philosophy.     

In vitro biosynthesis of juvenile hormone III by the corpora allata ofCalliphora vomitoria and its role in ovarian maturation and sexual receptivity

Summary JH III is the only JH detected by GLC-MS in medium from in vitro incubations of corpora allata of adult females ofCalliphora vomitoria. When corpora allata were removed from females at various times during the reproductive cycle and the JH III produced by the glands in vitro measured by a JH III radioimmunoassay, an increase in the level of synthesis was found to occur before previtellogenesis (0–24 h). A second increase appeared at the onset of vitellogenesis (72–83 h) and continued until the end of vitellogenesis (96 h) and the occurrence of chorionation (120 h). Since sexual receptivity develops with vitellogenesis, the significantly higher levels of JH III biosynthesis in vitro at this time supports a possible role for JH in the acquisitive of receptivity.     

Action of juvenile hormone on the follicle cells ofRhodnius prolixus: Evidence for a novel regulatory mechanism involving protein kinase C

Summary Juvenile hormone (JH) is known to act on the membranes of the follicle cells ofRhodnius, activating a specific Na⁺, K⁺-ATPase. This leads to a decrease in volume of the cells and the appearance of spaces between them (patency). The addition of an inhibitor of protein kinase C, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), to the medium in vitro inhibits the action of JH on the follicle cells. PDBU (phorbol-12,13-dibutyrate) mimics the action of JH in vitro and the response of the follicle cells to, PDBU is blocked by ouabain. It is concluded that the activation of protein kinase C is a required step in the chain of events leading to activation of the JH-dependent ATPase and set in train by the binding of JH to the membrane.     

A specific binding protein for the moulting hormone ecdysterone in locust haemolymph

Summary Specific binding of³H-ecdysterone to a high mol. wt. protein from Locusta migratoria haemolymph was shown by gel filtration. The hormone-protein complex shos a dissociation constant K_d3.10^–7 M, and the concentration of binding sites varies during the last larval instar.     

Flying insects: model systems in exercise physiology

Insect flight is the most energy-demanding exercise known. It requires very effective coupling of adenosine triphosphate (ATP) hydrolysis and regeneration in the working flight muscles.³¹P nuclear magnetic resonance (NMR) spectroscopy of locust flight muscle in vivo has shown that flight causes only a small decrease in the content of ATP, whereas the free concentrations of inorganic phosphate (P _i ), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) were estimated to increase by about 3-, 5- and 27-fold, respectively. These metabolites are potent activators of glycogen phosphorylase and phosphofructokinase (PFK). Activation of glycolysis by AMP and P _i is reinforced synergistically by fructose 2,6-bisphosphate (F2,6P₂), a very potent activator of PFK. During prolonged flight locusts gradually change from using carbohydrate to lipids as their main fuel. This requires a decrease in glycolytic flux which is brought about, at least in part, by a marked decrease in the content of F2,6P₂ in flight muscle (by 80% within 15 min of flight). The synthesis of F2,6P₂ in flight muscle can be stimulated by the nervous system via the biogenic amine octopamine. Octopamine and F2,6P₂ seem to be part of a mechanism to control the rate of carbohydrate oxidation in flight muscle and thus function in the metabolic integration of insect flight.Dedicated to Dr. Ernst Zebe, Emeritus Professor of Zoology (University of Münster) on the occasion of his 70th birthday.     

Transesterification of juvenile hormone occurs in vivo in locust when injected in alcoholic solvents

Catabolism of juvenile hormone was studied in vivo in the African locust by injection of the labelled natural enantiomer (10R) (12-³H) JH-III. Due to the poor solubility of JH in aqueous solution, it was injected in a water-miscible solvent. Ethanol was chosen for its apparently low toxicity towards the locust. In these experimental conditions, reverse phase liquid chromatography procedure (RP-HPLC) coupled with on line radiodetection, revealed an apolar metabolite of JH-III. This compound was found both in adult females and in fifth stadium larvae. We demonstrate that this metabolite resulted from substitution of the carboxyl methyl group of JH-III by some hydrophobic moiety. This compound co-migrates in our RP-HPLC system with the JH analog epoxy-ethyl farnesoate (JH-III ethyl ester) obtained by KCN-catalysed transesterification of JH-III in ethanol. Both JH-III ethyl ester of chemical origin and biological compounds extracted from locusts give the same spectra when analyzed by gas chromatography-mass spectroscopy (GC-MS). Transesterification of JH-III was not observed with locust tissues incubated in vitro but occurred in vivo even if JH was injected in other alcoholic solvents such as propanol. Our data suggest that transesterification of JH-III occurred in vivo and underline the role of injecting solvent in in vivo studies.     

Free amino acids of the haemolymph of the cotton leaf-worm,Spodoptera littoralis Boisduval full-grown larvae,infected with nuclear-polyhedrosis virus

Summary Free amino acids in the haemolymph ofSpodoptera littoralis full-grown larvae infected with a nuclearpolyhedrosis virus were compared with those in the haemolymph of normal insects. Amino acids were separated by 2-dimensional paper chromatography and quantified colorimetrically. Most of the amino acids in the haemolymph of diseased full-grown larvae decreased markedly in concentration but proline, lysine, aspartic acid and histidine occurred in greater concentration in the haemolymph of diseased full-grown larvae than in the haemolymph of healthy insects.     

Juvenile hormone titre and regulation in the cockroachDiploptera punctata

Summary Titres of juvenile hormone (JH) have been determined in both hemolymph and whole body extracts of femaleDiploptera punctata during the first gonotrophic cycle using a method employing gas chromatography/mass spectrometry for qualitative and quantitative analysis. JH III is the sole JH found in both adult and last instarD. punctata. Maximum values of 1500 ng/ml (6M) were observed at the middle of the gonotrophic cycle, when basal oocyte growth rate was greatest. Changes in rates of JH release in vitro by corpora allata paralleled closely the changes in JH titre, suggesting that biosynthesis is a major regulator of titre. JH levels per animal were calculated from observed JH titres, and at certain time points in the gonotrophic cycle JH levels obtained from analysis of whole bodies were significantly greater than those predicted from hemolymph titres. These results suggest the existence of a nonhemolymph JH pool inD. punctata. Decay in JH titre after allatectomy of 5 day females has also been studied. Following a rapid initial decline, the rate of decay slowed appreciably 4 h post-operation. Thus, use of a first-order rate constant to estimate half-life of JH significantly underestimated the longevity of the hormone. The apparent persistence of JH following allatectomy may be due to the existence of a nonhemolymph JH pool.     

The regulation of trehalose metabolism in insects

Trehalose is a non-reducing disaccharide comprising two glucose molecules. It is present in high concentration as the main haemolymph (blood) sugar in insects. The synthesis of trehalose in the fat body (an organ analogous in function to a combination of liver and adipose tissue in vertebrates) is stimulated by neuropeptides (hypertrehalosaemic hormones), released from the corpora cardiaca, a neurohaemal organ associated with the brain. The peptides cause a decrease in the content of fructose 2,6-bisphosphate in fat body cells. Fructose 2,6-bisphosphate, acting synergistically with AMP, is a potent activator of the glycolytic enzyme 6-phosphofructokinase-1 and a strong inhibitor of the gluconeogenic enzyme fructose 1,6-bisphosphatase. This indicates that fructose 2,6-bisphosphate is a key metabolic signal in the regulation of trehalose synthesis in insects. Trehalose is hydrolysed by trehalase (E.C. 3.2.1.28). The activity of this enzyme is regulated in flight muscle, but the mechanism by which this is achieved is unknown. Trehalase from locust, flight muscle is a glycoprotein bound to membranes of the microsomal fraction. The enzyme can be activated by detergents in vitro and by short flight intervals in vivo, which indicates that changes in the membrane environment modulate trehalase activity under physiological conditions.     

Relationship between the absolute configuration and the biological activity of juvenile hormone III

Summary The activity of the pure 10R (=natural) and 10S enantiomers of juvenile hormone III (JH III) was determined in 3 different bioassays, and the relative binding affinity of the 2 enantiomers to the haemolymph JH-binding protein of the cockroachNauphoeta cinerea was measured. In theGalleria wax test, a local morphogenetic assay, the 10R enantiomer was 5240 times more active than, the 10S enantiomer, 1Galleria unit corresponding to 0.42 pg of 10R-JH III as compared to 2.2 ng for 10S-JH III. In a systemic morphogenetic assay with the cockroachNauphoeta cinerea 380 times less 10R enantiomer was necessary in order to induce detectable juvenilisation (58 ng 10R and 22 g 10S) and in a systemic gonadotropic assay withNauphoeta cinerea 255 times less 10R was needed to induce vitellogenin synthesis in 50% of the insects (6.7 ng 10R and 1710 ng 10S). In the JH-binding protein assay 10R-JH III had an affinity for the JH-binding protein (lipophorin) which was approximately 46 times higher than that of 10S-JH III.     

Culturein vitro d'un tissu nymphal de lépidoptère

Summary Using a simplified medium, the composition of which is described in the text, cell cultures (fibroblast type) were obtained from nymphal ovarian tubes ofBombyx mori. The change in the form between fibroblasts and spindle cells is described. The preservation of activity of haemolymph congealed to – 20° C for a long time is shown; the advantages of using nymphs for cultivating tissues in virological series are also noted.     

Changes in protein-calcium association during different hours of a day in the haemolymph of the crabScylla serrata (Forskal)

Summary InScylla serrata, the haemolymph free calcium and bound calcium showed fluctuations during the different hours of the day. Whenever bound calcium decreased or increased in the haemolymph, the free calcium increased or decreased respectively; the significance of this is discussed. Besides, the suitability and reliability of Webster's chloranilic acid method, compared with other methods for the determination of haemolymph free and bound calcium, are empirically assessed.     

Forced synthesis of trace amounts of juvenile hormone II from propionate by corpora allata of a juvenile hormone III-producing insect

Summary Corpora allata of the cockroachDiploptera punctata normally synthesize only the isoprenoid juvenile hormone III (JH III). Only under extreme in vitro conditions (absence of carbon sources other than propionate) do they produce trace amounts of the homoisoprenoid JH II in addition to JH III. The specificity of the in vitro synthesis of JH III byD. punctata is thus consistent with the observed lack of homoisoprenoid JHs in this insect.     

Juvenile hormone I is the principal juvenile hormone in a hemipteran insect,Riptortus clavatus

Juvenile hormone I (JH I) was identified by combined gas chromatography/mass spectrometry as the predominant JH in the hemolymph of female adults of the bean bug,Riptortus clavatus (Thunberg) (Hemiptera: Alydidae). Among JH I, II, and III, JH I was the most effective hormone for inducing the synthesis of yolk proteins in diapause adults.     

Juvenile hormone titre and regulation in the cockroach Diploptera punctata

Titres of juvenile hormone (JH) have been determined in both hemolymph and whole body extracts of female Diploptera punctata during the first gonotrophic cycle using a method employing gas chromatography/mass spectrometry for qualitative and quantitative analysis. JH III is the sole JH found in both adult and last instar D. punctata. Maximum values of approximately 1500 ng/ml (approximately 6 microM) were observed at the middle of the gonotrophic cycle, when basal oocyte growth rate was greatest. Changes in rates of JH release in vitro by corpora allata paralleled closely the changes in JH titre, suggesting that biosynthesis is a major regulator of titre. JH levels per animal were calculated from observed JH titres, and at certain time points in the gonotrophic cycle JH obtained from analysis of whole bodies were significantly greater than those predicted from hemolymph titres. These results suggest the existence of a nonhemolymph JH pool in D. punctata. Decay in JH titre after allatectomy of 5 day females has also been studied. Following a rapid initial decline, the rate of decay slowed appreciably 4 h post-operation. Thus, use of a first-order rate constant to estimate half-life of JH significantly underestimated the longevity of the hormone. The apparent persistence of JH following allatectomy may be due to the existence of a nonhemolymph JH pool.