Insect anti-juvenile hormone and juvenile hormone activity from plants in the genusNama

Summary The insect anti-juvenile hormones precocene I and II (7-methoxy-2,2-dimethyl-2H-1-benzopyran and 6,7-dimethoxy-2,2-dimethyl-2H-1-benzopyran) were identified in three of nineNama (Hydrophyllaceae) species. Precocene I occurred inN. lobbii while precocene II occurred inN. hispidum, N. lobbii andN. sandwicense. N. hispidum contained the highest concentration (ca 0.5% dry weight) of precocene II, which was found in the leaves, stems, seed capsules, corolla, glandular trichomes, and seeds. In addition to the anti-juvenile hormone, insect juvenile hormone activity was detected in the organosoluble extracts ofN. rothrockii andN. sandwicense. N. sandwicense is the first plant discovered to contain compounds with both anti- and juvenile hormone activity.     

Effects of methoprene and juvenile hormone on the oxidative metabolism of isolated mitochondria from flight muscle ofLocusta migratoria L.

Summary In vitro applications of juvenile hormone III and a juvenile hormone analogue, methoprene, were made to mitochondria isolated from dorsal longitudinal flight muscles of adultLocusta migratoria L. Both compounds completely inhibited oxygen consumption at the highest concentrations used. At lower concentrations, state 3 respiration and respiratory control were reduced but the ADP/O ratio was largely unaffected.     

Cycles of juvenile hormone esterase activity during the juvenile hormone-driven cycles of oxygen consumption in pupal diapause of flesh flies

Summary During diapause O₂ consumption in fly pupae is a cyclic event (4-day periodicity at 25°C) driven by cycles of juvenile hormone activity. Levels of juvenile hormone esterase activity change systematically during the cycle, with highest activity observed at the nadir of the O₂ consumption cycle.Supported in part by grant 88-37153-3473 from the USDA Competitive Grants Office and grant 23-088 from the USDA Forest Service. Thanks to Dr Ming Tu Chang for her helpful advice.     

Head critical period and juvenile hormone titre during the last larval instar of the cockroach,Periplaneta americana

From neck ligation experiments with last instar larvae of the cockroachPeriplaneta americana it was concluded that the head critical period is reached around day 17, which corresponds to 59% of the last larval stage. At the same stage the juvenile hormone III titre in the hemolymph dropped to undectable levels.     

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.     

Titers of ecdysone, 20-hydroxyecdysone and juvenile hormone III throughout the life cycle of a hemimetabolous insect,the ovoviviparous cockroachNauphoeta cinerea

Summary Titers of ecdysone, 20-hydroxyecdysone and juvenile hormone III were measured in whole body extracts or hemolymph of embryos, first, penultimate and last stadium nymphs, and adult females ofNaupoheta cinerea. We used a gas-chromatography/mass spectrometry method for quantifying juvenile hormone and a radio-immunoassay for ecdysteroid determination. Juvenile hormone III is particularly abundant in the embryonic stage (up to 960 ng/g), at a low level in first and penultimate stadium nymphs (2–10 ng/ml) and almost absent in the last nymphal stadium; in the adult female the juvenile hormone titer rises to 180 ng/ml in hemolymph during rapid oocyte growth. The titers of ecdysone and 20-hydroxyecdysone undergo similar fluctuations in the embryonic and nymphal stages, being highest at the time of cuticle formation in the embryo and a few days before the nymphal and adult molts (around 100–200 ng/ml for exdysone and 2–4 g/ml for 20-hydroxyecdysone).Acknowledgments. We thank Mrs A. Tschan for rearing the cockroaches, Mr M. Kaltenrieder for drawing the graphs, Mr G.C. Jamieson and Mrs C. Reuter for GC/MS analyses. We are also grateful to the Swiss National Science Foundation (grant no. 3.291-0.82 to B. Lanzrein) and the United States National Science Foundation (grant no. PCM 82-08665 to D.A. Schooley) for their financial support.     

Phosphoramidon enhances allatostatin-mediated inhibition of juvenile hormone biosynthesis in the corpora allta of the cockroach,Diploptera punctata

Use of the enkephalinase inhibitor phosphoramidon in the in vitro radiochemical assay for juvenile hormone biosynthesis enhanced allatostatin-mediated inhibition of hormone production by corpora allata of the cockroach,Diploptera punctata. Significant increases in inhibition in day 2 virgin female CA by AST 1 (at 10^–7 M) and AST 4 (10^–8–10^–7 M) were observed in the presence of phosphoramidon (10^–5M or greater). No significant increases in inhibition were seen in CA from day 6 mated females with AST 4 (10^–9–10^–7M) and phosphoramidon combined. Phosphoramidon alone had no effect on JH biosynthesis. Analysis of allatostatin content of the CA, as determined by ELISA, revealed that addition of phosphoramidon to the medium increased the endogenous allatostatin conten in CA of virgin and mated females. The similarity in primary structure between allatostatins and enkephalin-like peptides and their similar distribution makes it probable that phosphoramidon acts by preventing breakdown of allatostatins within the CA.     

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.     

Effects of a juvenile hormone mimetic,fenoxycarb, on post-embryonic development of the European corn borer,Ostrinia nubilalis Hbn.

Summary The effect of a juvenile hormone mimetic, fenoxycarb, Ro 13-5223, was tested on the larval instars of the European corn borer,Ostrinia nubilalis, by dipping or topical application. When larvae were treated in instars 2, 3 or 4, the duration of the fifth instar was modified. More permanent and fewer supernumerary larvae were obtained when treatment occurred in the early instars. This non-neurotoxic compound exhibited a strong dose-dependent juvenile hormone type of activity when it was applied to last instar larvae. Fenoxycarb prevented the onset of pupation and produced supernumerary larvae and intermediates. Permanent larvae were obtained if fenoxycarb was applied on day 0 or day 1 of the last instar. The use of such a JH mimetic in the understanding of endocrine control of diapause is discussed.     

Immunochemical similarities among the hemolymph juvenile hormone binding proteins of moths

The hemolymph from various species of moths was analyzed for cross-reactivity with a panel of six monoclonal antibodies made against the hemolymph juvenile hormone binding protein ofManduca sexta. With the exception of one antibody, the immunoreactivity was limited to the sphingid family. One monoclonal antibody cross-reacted with a number of lepidopteran species; however, families such as Noctuidae and Pyralidae, known to have high affinity, low molecular weight juvenile hormone binding proteins, did not cross-react. Immunological cross-reactivity withManduca sexta juvenile hormone binding protein in several primitive moth families supports the current model of phylogenetic relationships in the order Lepidoptera.     

Autoinhibition of juvenile hormone production. The case of the cockroachBlattella germanica (L.)

In 6-day-old females ofBlattella germanica, the activity of corpora allata (CA) was inhibited in vitro by juvenile hormone III (JH III). Effective doses (281.5 and 375.4 M in the medium) were somewhat higher than (although of the same order of magnitude as) the estimated intraglandular concentration of JH III at this age, and they induced about 45% inhibition of hormonal release and a significant intraglandular accumulation of JH III and methyl farnesoate. The results suggest that autoinhibitory mechanisms operate in the CA to constrain the upper limit of JH III production at the end of the gonadotrophic cycle.     

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.     

Biological activity of enantiomerically pure forms of insect juvenile hormone I and III inBombyx mori

Summary Biological activity of enantiomerically pure juvenile hormones was assayed by topical application on allatectomizedBombyx fourth instar larvae. JHs tested were (10R)-JH I [methyl (2E,6E,10R,11S)-10,11-epoxy-3,11-dimethyl-7-ethyl-2,6-tridecadienoate], (10S)-JH I [methyl (2E, 6E, 10S, 11R)-10,11-epoxy-3,11-dimethyl-7-ethyl-2,6-tridecadienoate], (10R)-JH III [methyl (2E,6E,10R)-10,11-epoxy-3,7,11-trimethyl-2,6-dodecadienoate] and (10S)-JH III [methyl (2E,6E,10S)-10,11-epoxy-3,7,11-trimethyl-2,6-dodecadienoate]. Among these compounds, natural (10R)-JH I was most active and the dose needed to induce 50% larval molting was 0.04 g/larva; it was approximately 12,000 times more active than unnatural (10S)-JH I. Though natural (10R)-JH III showed slight biological activity, it was only one three-thousandth of that of (10R)-JH I. Unnatural (10S)-JH III exhibited no biological activity at the levels assayed.     

Differences in the stimulation by calcium ionophore of juvenile hormone III release from corpora allata of solitarious and gregariousLocusta migratoria

Summary Incubation of the calcium ionophore A23187 resulted in an increase in the median rate of juvenile hormone III release by corpora allata (CA) of both gregarious and solitarious adultLocusta migratoria females at 3, 5 and 8 days after fledging. At all 3 datapoints, the enhancement of release rates was highly significant for CA from gregarious females but not significant for CA from solitarious females.     

Juvenile hormone degradation in brain and corpora cardiaca-corpora allata complex during the last larval instar ofGalleria mellonela (Lepidoptera,Pyralidae)

Summary Time course analysis of juvenile hormone degradation in the brain and the corpora cardiaca-corpora allata complex shows that during the first two days of the last larval instar the juvenile hormone degradation is very low. Starting from the third day up to the seventh day a continuous increase of esterase activity is observed.     

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.     

Repair of integumental lesions in the decapod crab,Carcinus maenas (L.), in the presence of molting hormone

Summary Epicuticle is not included as any part of the repair cuticle that closes integumental lesions in crabs receiving multiple injections of molting hormone (20-hydroxyecdysone) before and/or after damage to the integument. It is suggested that deposition of the exclusively lamellate material of repair cuticle is mediated by some epidermal factor(s) rather than by external, humoral influences.