In vitro inactivation of corpora allata of the bugOncopeltus fasciatus by precocene II

Summary Corpora allata fromOncopeltus fasciatus incubated in vitro in medium containing 10^–5.35 M (1 g/ml) of precocene II lose their ability to secrete juvenile hormone when reimplanted into last instar larvae.Acknowledgments. We thank Mr K. Dorn, Mrs L. Dolezal, Mrs V. Nötzli-Graf, Mr K.H. Trautmann and Mr A. Schuler for technical help, Dr W. Vogel and Dr A. Dübendorfer for valuable discussions.     

Effects of exogenous juvenile hormone I on the numbers and distribution of antennal and maxillary and labial palp sensilla of maleBlattella germanica (L.) (Dictyoptera: Blattellidae)

Summary Unlike in the female, application of exogenous juvenile hormone I to 6th-instar male nymphs does not result in the retention of nymphal characteristics on the antennae and maxillary and labial palps. JH-treatment differentially affects the numbers of sensilla on the 3 appendages of adult maleB. germanica.     

Types of response of insects on treatment with juvenile hormone active insect growth regulators

Summary The reactions of immature insects of more than 40 species placed in continuous contact with high doses of juvenile hormone active insect growth regulators were analyzed. 4 different types were recognized: Inhibition of both ecdysis and metamorphosis, defective metamorphosis, defective adult emergence and defective embryogenesis. The reactions are understood as a consequence of JH/ecdysone antagonism. The limits for the practical application of these substances are discussed.Acknowledgments. We wish to express our thanks to Miss M. Kälin, Miss M. Lehner, Mrs T. Mühle and Mr P. Müller for technical assistance and to Prof. G. Benz, Mr M. Gruber, Dr E. Günthart and Prof H. A. Schneiderman for helpful discussion and correction of the text.     

Ecdysteroid titers during larval reproduction of the dipteran insectHeteropeza pygmae

Summary Ecdysteroid titers were measured in whole-body extracts of pedogenetically reproducing larvae of the dipteran insectHeteropeza pygmaea and in the dietary fungus. The titers are very low in the first 3 days of larval growth, but increase during the last 2 days. The level of 20-hydroxyecdysone is then significantly higher than that of ecdysone. Measurements of the titers in the fungus gave no conclusive results.Supported by the Swiss National Science Foundation, grants No. 3.218-0.77 to D.F.W. and 3.714-0.80 to B.L. We thank Xiang-Xiong Zhu for help with the RIA and Mrs G. Rhyner for technical assistance. Gifts of ecdysone antiserum from Dr J. D. O'Connor and of 23,24-[³H₂]-ecdysone from Dr J. Koolman are also gratefully acknowledged.     

Ecdysteroid receptors in the neuroendocrine-endocrine axis of a moth

With a combination of thaw-mount autoradiography using a tritiated 20-hydroxyecdysone agonist, ponasterone A, and immunocytochemistry with a monoclonal antibody to 29 K-prothoracicotropic hormone, high affinity binding sites for ecdysteroids were identified in the tissues of the neuroendocrine-endocrine axis inManduca sexta larvae. At specific times during larval-pupal development in fifth stadium larvae, nuclear ecdysteroid binding sites were present in the cerebral prothoracicotropes, the corpora allata and prothoracic glands, the main axis for the regulation and production of ecdysteroids. A stage-specific appearance of ecdysteroid receptors also occurred in cells of fat body, midgut and Malpighian tubules, tissues which convert ecdysone into 20-hydroxyecdysone. Our data identify new target tissues for ecdysteroids and suggest that ecdysteroids could affect their own production at the genomic level via long and short feedback loops.     

RH-5849, a nonsteroidal ecdysone agonist,does not mimic makisterone-A inDysdercus koenigii

The response of final instar nymphs ofDysdercus koenigii to topical application of the non-steroidal ecdysone agonist, RH-5849, was dose dependent. The candidate compound produced mortality even at moderate doses, but precocious adult development was not observed. Similar results were obtained after oral administration or injection. Conversely, injections of makisterone-A (the principal moulting hormone ofDysdercus) into 5th instar nymphs resulted in precocious adult development within 4 days. We conclude that RH-5849 does not mimic makisterone-A, as is the case with ecdysone, and that toxicity is mediated instead through non-endocrine targets in this insect species.     

Sexual behavior without adult morphogenesis inLocusta migratoria

Summary Injection of azadirachtin into young Vth (=last) instarLocusta hoppers inhibits molt and a considerable portion of the animals survice for more than 40 days as over-aged nymphs; in contrast, the duration of the Vth instar is 8–10 days in normal controls. Males of over-aged nymphs exhibit adult sexual behavior, and injection of juvenile hormone intensifies this behavior. The results demonstrate that the terminal molt leading to morphogenetic adult differentiation is not necessary for an adult behavioral pattern to develop, and/or to become overt, in a hemimetabolous insect.We thank Prof. H. Rembold for a generous gift of azadirachtin and Mr Y. Toren for keeping the locust culture.     

Distribution of free and conjugated ecdysteroids between follicle cell sheath and ooplasm in oocytes of the cockroachNauphoeta cinerea

Summary It is shown that both follicle cell sheath and ooplasm contain only small quantities ofHelix-hydrolyzable ecdysteroid conjugates and that 20-hydroxy-ecdysone is the predominant free ecdysteroid. Its concentration is very high in the follicle cell sheath immediately before chorion formation (6825 ng/g) and much lower after chorion formation (150 ng/g), while it is 234 ng/g and 95 ng/g in the ooplasm at the same stages respectively.Thanks are due to Dr J.D. O'Connor (Los Angeles) for the ecdysone antiserum, to Dr J. Koolman (Marburg) for 23,24-[³H]-ecdysone and to Mrs A. Tschan for technical assistance. A fellowship to X. Zhu from the Chinese Ministry of Education and financial support from the Swiss National Science Foundation (grant no. 3.714-0.80 to B. Lanzrein) are gratefully acknowledged.     

Role of low ecdysteroid levels in the early last larval instar ofBombyx mori

The relationship between juvenile hormone and ecdysteroid titers during the early stage of the last larval instar of the silkworm,Bombyx mori, was examined in this study. When larvae were fed 20-hydroxyecdysone-supplemented mulberry leaves throughout the last larval instar, 100% underwent supernumerary larval molting instead of metamorphosis. The application of juvenile hormone mimic during the early last instar did not induce supernumerary larval molting, but did delay metamorphosis. Temporal and quantitative ecdysteroid titer measurements revealed that in normal larvae the titers maintained very low levels (3–12 ng/ml) during the early stage of the last instar; however, in 20-hydroxyecdysone-treated larvae, levels ranging from 24 to 45 ng/ml were detected, and a major peak (246 ng/ml) was observed on day 6. These results show that very low ecdysteroid titer levels during the early stage of the last larval instar may play an important role in initiating decreases in juvenile hormone titers as well as in directing metamorphosis.     

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.     

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.     

Pancreatic polypeptide: A possible role in the regulation of food intake in the mouse. Hypothesis

Summary Pancreatic polypeptide (PP) is a recently identified hormone produced by pancreatic endocrine cells. The islets of genetically obese mice (ob/ob, C57 BL/6J), which are suspected to lack a circulating satiety factor, contain relatively few of the PP-producing cells. Administration of bovine pancreatic polypeptide (bPP) reduces food intake and suppresses body weight gain in the hyperphagic obese mice. It is postulated that PP participates in the regulation of food intake in a manner as yet undefined.This work was supported by grant No. 3.553.75 from Swiss National Science Foundation. We thank Mrs M. Eissler and Mr R. Cuche for their valuable help.     

Ecdysteroids: possible candidates for the hormone which triggers salivary gland degeneration in the ixodid tickAmblyomma hebraeum

Summary Following engorgement, female ixodid ticks secrete a tick salivary gland degeneration factor (TSGDF) into the hemolymph. Here we show that the arthropod ecdysteroid hormones, ecdysome and 20-hydroxyecdysone, induce degeneration of tick salivary glands maintained in organ culture. The effective dose range in vitro is 30–300 ng/ml, a range reported to be physiological for this species following repletion. In addition, infusion of 20-hydroxyecdysome in vivo induces salivary gland degeneration. We therefore propose that TSGDF may be an ecdysteroid.Acknowledgments. Some of the data reported here were presented to the annual meeting of the Canadian Society of Zoologists, 15–18 May 1983; Program of abstracts, page 53. Financial support of the Canadian National Sportsmen's Fund and NSERC Canada to W.R.K. is gratefully acknowledged.     

Freeze tolerance in the frog,Rana sylvatica

Summary Wood frogs survive extracellular freezing at moderate subzero temperatures (–4°C) for at least 11 days. Freezing survival is aided by the accumulation of high concentrations of glucose as a cryoprotectant in blood and tissues. Glucose production was accompanied by a rapid decline in liver, but not muscle, glycogen levels suggesting that liver is the organ controlling cryoprotectant synthesis.Acknowledgments. This study was supported by grants to L.M.G. from the Kroc Foundation (Santa Ynez, California) and from the National Institute of Dental Research (Grant No. DE-03987). The authors wish to thank K. Yorko, M. Shakin, J. Finan and Mrs N. Manivannan for their technical and secretarial assistance.Acknowledgments. I thank Dr J. Ballantyne, Dr F. Schueler and I. McMurray for help with frog collections and Dr. W. Schmid, Dr J. Bogart and Dr F. Cook for helpful discussions. Supported by an N.S.E.R.C. operating grant and by a grant from the Atkinson Charitable Foundation.     

Carboxylesterases of high molecular weight in the hemolymph ofLocusta migratoria

Summary The main carboxylesterase in the hemolymph of the migratory locust,Locusta migratoria, is a protein of high molecular weight; about 700–750 kDa. This esterase hydrolyzes juvenile hormone III, -naphthylacetate and -naphthylacetate. The carboxylesterase dissociates to give an esterase of molecular weight 148 kDa after treatment of the hemolymph with mercaptoethanol.     

Critical period and ecdysone titers in the pupae ofPieris brassicae

Summary The ecdysone titer in the haemolymph of pupae of Pieris brassicae shows a sharp peak (1.90 g/ml) on days 5 and 6 of the pupal stage. Isolated abdomens were able to form adult abdomens at 72 h, this being 2 or 3 days before maximum ecdysone levels and at a time when the concentration of hormone in the abdominal haemolymph was only 0.23 g/ml.     

Effect of air ionization on blood serotonin in vitro

Summary The effect of negative and positive air ionisation on siliconized blood serotonin was studied in vitro. The experiments showed that within 10 min positive ionisation increased serotonin levels in total blood (+40%), plasma (+90%), erythrocytes (+50%) and thrombocytes (+200%). On the other hand, negative ionization (10 min) lowered the serotonin content of total blood (–30%), plasma (–42.5%), erythrocytes (–41.7%) and thrombocytes (–72.3%), thus confirming the Krueger Effect in vitro.This research was generously supported by a grant from the Joint Research Fund of the Hebrew University and Hadassah, and Dr. and Mrs.Leonard Appleby of Palmbeach, Florida.Acknowledgment: Our thanks are due to Mr.B. Shalita, Mrs.Zippora Adar and Mrs.Olga Fradkin for devoted technical assistance.     

In vitro secretion of ecdysteroids by Y-organs of the crayfish,Procambarus clarkii

It was demonstrated that excised Y-organs of the crayfish,Procambarus clarkii, synthesize in vitro 3-dehydroecdysone (3-DHE) as the major product, together with small amounts of ecdysone. Both were identified by immunological and spectroscopic methods. The increase of ecdysteroidogenesis in the Y-organs was accompanied by an increase of the major free ecdysteroid, 20-hydroxyecdysone, in the hemolymph. This suggests a physiological role of 3-DHE, the details of which are still to be elucidated.     

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.     

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.