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.     

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.     

Delayed effects of juvenile hormone analogues on metamorphosis ofPyrrhocoris apterus are not mediated by the corpus allatum

Summary While it is clear that juvenile hormone analogues disturb metamorphosis, there is some controversy about their effect on the activity of the corpus allatum in the course of metamorphosis. The present experiments showed that juvenile hormone analogues applied to eggs did not prevent inactivation of the corpus allatum at the onset of the last larval instar ofPyrrhocoris apterus.     

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.     

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.     

Ecdysteroid-dependent larval-adult oviduct transformation in the milkweed bugOncopeltus fasciatus requires absence of juvenile hormone

Summary It has been tested whether juvenile hormone plays a role in the larval-adult transformation of lateral oviducts in the milkweed bug. The transformation is ecdysteroid-dependent, as was reported previously². Application of precocene or juvenile hormone III proved that the absence of juvenile hormone is required.Acknowledgments. This work was supported by a grant (Do 163/91) of the DFG. We thank Ms C. Friederichs for excellent technical support.     

Reactivation of juvenile hormone synthesis in adult drones of the honey bee,Apis mellifera carnica

Juvenile hormone synthesis was measured by the corpora allata radiochemical assay in vitro. No hormone was produced during pupal stages, but soon after adult eclosion the corpora allata were reactived. The rate of juvenile hormone III synthesis increased until day 10 after emergence. Possible functions of juvenile hormone in adult drones are 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.     

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.     

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.     

Reversible juvenile hormone inhibition of ecdysteroid and juvenile hormone synthesis by the ring gland of Drosophila melanogaster

Juvenile hormone bisepoxide (JHB3) and juvenile hormone III (JH III) both inhibited the in vitro production of ecdysteroids by ring glands and brain-ring gland complexes from third instar post-feeding larvae of Drosophila melanogaster in a reversible manner, although JHB3 had greater efficacy. The JH III and JHB3 precursor, methyl farnesoate, did not affect ecdysteroid production. The in vitro synthesis of total detectable JH (JHB3 + JH III + methyl farnesoate) by the corpus allatum portion of the isolated ring gland was also inhibited reversibly in the presence of exogenous JHB3 and JH III, but not by methyl farnesoate. These data indicating negative feedback are in agreement with the accepted dogma of endocrine gland regulation.     

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

Corpora allata of the cockroach Diploptera 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 by D. punctata is thus consistent with the observed lack of homoisoprenoid JHs in this insect.     

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.     

Effective dose present in cockroach larvae exposed continuously to a juvenile hormone active insect growth regulator

Summary Larvae of the German cockroach exposed to filter paper impregnated with juvenile hormone (JH) active substance contain, at the middle of the last instar, about one-hundredth of the dose applied to 1 cm². The amount of metabolized substance rises sharply before and disappears rapidly after the ecdysis into the supernumerary instar.Acknowledgments. We wish to express our thanks to Prof. Dr H.A. Schneiderman, Dr W. Vogel and Dr P.A. Vonder Mühll for stimulating discussion and to Mr M. Gruber for correction of the text.     

Reversible juvenile hormone inhibition of ecdysteroid and juvenile hormone synthesis by the ring gland ofDrosophila melanogaster

Juvenile hormone bisepoxide (JHB₃) and juvenile hormone III (JH III) both inhibited the in vitro production of ecdysteroids by ring glands and brain-ring gland complexes from third instar post-feeding larvae ofDrosophila melanogaster in a reversible manner, although JHB₃ had greater efficacy. The JH III and JHB₃ precursor, methyl farnesoate, did not affect ecdysteroid production. The in vitro synthesis of total detectable JH (JHB₃+JH III+methyl farnesoate) by the corpus allatum portion of the isolated ring gland was also inhibited reversibly in the presence of exogenous JHB₃ and JH III, but not by methyl farnesoate. These data indicating negative feedback are in agreement with the accepted dogma of endocrine gland regulation.     

The effects of juvenile hormone analogues on the eggs ofPieris brassicae L.

Summary Mortality was produced in the eggs ofP. brassicae after treatment with 3 juvenile hormone analogues. The mortality was greatest in the early stages of embryonic development.I am grateful to the United Kingdom Ministry of Agriculture, Fisheries and Food, who provided a Research Studentship for this work.     

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.     

Preparation of optically pure juvenile hormone I labelled in the ester methyl group with tritium at very high specific activity

Summary Radiolabelled juvenile hormone I (3) was prepared in an optically pure form by an enzymatic method using the capacity of accessory sex gland homogenates from maleHyalophora cecropia to transfer the methyl group of S-adenosyl-L-methionine to the carboxylic acid function of the (10R, 11S)-enantiomer of racemic presursor2; the specific activity of3 is higher than 53 Ci/mmole (1.96 TBq/mmole).We are indebted to Dr W. Thelfer, Philadelphia, for providing us with diapausing pupae of male Cecropia silk moths, and to Dipl.-Biol. G. Klages, Darmstadt, for preparing racemic JH-Iacid. Financial support by the Deutsche Forschungsgemeinschaft (grant No. Em 14/8) is gratefully acknowledged.     

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.     

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

Use of 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(-5) M or greater). No significant increase in inhibition were seen in CA from day 6 mated females with AST 4 (10(-9)-10(-7) M) 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 content 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.