The activity of juvenile hormone mimics for the eggs of Rhodnius prolixus

Twenty four juvenile hormone (JH) mimics of several different chemical classes were applied to freshly laid eggs of Rhodnius prolixus. Activity of the JH mimics was assessed in terms of their ability to inhibit eclosion. All chemical classes tested except one of the natural JHs and two very closely related long chain compounds contained members which inhibited eclosion; the most active chemicals were derivatives of geranyl para-aminobenzoic acid esters and aryl methylene dioxyphenyl ethers. In general, those chemicals of a particular class which had highest morphogenetic activity when applied to last instar larvae, were also most active in inhibiting eclosion. However, not all structure/activity relationships of JH mimics for last instar larvae were applicable to eggs; in particular the presence of a terminal epoxide group increased activity greatly for eggs but only slightly for last instar larvae. The dose per unit weight to inhibit eclosion is far greater than the equivalent dose to disrupt normal metamorphosis.     

Inhibitors of ester hydrolysis as synergists for biological activity of Cecropia juvenile hormone.

In Manduca sexta pupae the sensitivity to exogenous Cecropia C18 juvenile hormone (JH) is governed in large part by the rate at which the hormone is metabolized. By analysing the distribution of labelled JH and its metabolites, when dispensed by injection in light mineral oil, I have shown that the half-life of JH at concentrations ranging from 0·02 to 1 μg/g live weight of insect is approximately 24 hr.When the same doses of JH are dispensed in mineral oil containing endocrinologically inactive carboxylesterase inhibitors, organophosphate, and carbamate insecticides, the hormone's half-life in certain cases is prolonged significantly. The formulations containing those materials which most effectively prolong the hormone's half-life give higher assay scores than equivalent doses of hormone in mineral oil alone.From these results it is inferred that the chemicals tested synergize the Cecropia juvenile hormone's biological activity by serving presumably as inhibitors of the principal enzyme responsible for the rapid breakdown of exogenous C18-JH in haemolymph—a carboxylesterase. Preliminary in vitro experiments with carboxylesterase-rich Manduca pupal blood further support this inference.     

The effect of the persistence of juvenile hormone mimics on their activity on Rhodnius prolixus

The apparent juvenile hormone (JH) activity of methyl 12-homo-juvenate on fed fifth instar Rhodnius prolixus is nearly twelve times greater if the compound is applied topically over a period of 7 days (serial application) rather than as a single dose 1 day after feeding. Of five other JH mimics tested, four were more active by serial application, although none showed so great an increase in activity as methyl 12-homojuvenate; only farnesyl methyl ether was more active by single application. It is considered that the generally greater activity shown in the serial application tests is because less JH mimic is wasted by degradation and/or excretion. Therefore the serial application test provides a more accurate estimate of inna e JH activity than the single application test.     

A comparison of the morphogenetic and sterilizing activities of juvenile hormone mimics on Aedes aegypti

At 25°C, adult female aedes aegypti are most sensitive to sterilization by juvenile hormone (JH) mimics when such chemicals are applied 32 to 36 hr after the blood meal (when the ovaries are at late stage III to early stage IV). Application of JH mimics during this period reduces egg fertility and female fecundity and induces the production of large numbers of visually abnormal eggs. As the most sensitive phase for sterilization with JH mimics is well before oviposition, and as many abnormal eggs are laid following JH mimic treatment, it is likely that in this species sterilization effects are induced by some action on the developing oöcyte rather than on embryonic development.The relative activities of several JH mimics in sterilizing adult female A. aegypti are very similar to their relative activities in inhibiting metamorphosis. Thus the sterilizing action of JH mimics is likely to be a true JH effect and can be used as a test for JH activity for A. aegypti.     

Molecular similarity analysis between insect juvenile hormone and N, N-diethyl-m-toluamide (DEET) analogs may aid design of novel insect repellents

Molecular similarity analysis of stereoelectronic properties between natural insect juvenile hormone (JH), -a synthetic insect juvenile hormone mimic (JH-mimic, undecen-2-yl carbamate), and N, N-diethyl-m-toluamide (DEET) and its analogs reveals similarities that may aid the design of more efficacious insect repellents and give a better insight into the mechanism of repellent action. The study involves quantum chemical calculations using the AM1 semi-empirical computational method enabling a conformational search for the lowest and most abundant energy conformers of JH, JH-mimic, and 15 DEET compounds, followed by complete geometry optimization of the conformers. Similarity analyses of stereoelectronic properties such as structural parameters, atomic charges, dipole moments, molecular electrostatic potentials, and highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energies were performed on JH, JH-mimic and the DEET compounds. The similarity of stereoelectronic attributes of the amide/ester moiety, the negative electrostatic potential regions beyond the van der Waals surface, and the large distribution of hydrophobic regions in the compounds appear to be the three important factors leading to a similar interaction with the JH receptor. The similarity of electrostatic profiles beyond the van der Waals surface is likely to play a crucial role in molecular recognition interaction with the JH receptor from a distance. This also suggests electrostatic bioisosterism of the amide group of the DEET compounds and JH-mimic and, thus, a model for molecular recognition at the JH receptor. The insect repellent property of the DEET analogs may thus be attributed to a conflict of complementarity for the JH receptor binding sites.     

Mosquito larvicidal activities of farnesol and farnesyl acetate via regulation of juvenile hormone receptor complex formation in Aedes mosquito

Due to their target specificity and low-toxicity, insect growth regulators (IGRs) are regarded as promising alternatives to chemical insecticides. In this study, farnesol and farnesyl acetate exhibited juvenile hormone (JH)-based IGR activities. While farnesyl acetate showed JH agonist (JHA) activity in concentration-dependent manner, farnesol was identified as JH antagonist (JHAN) by interfering pyriproxyfen-mediated binding JH receptor complex. Both compounds showed mosquito larvicidal activities and caused retardation of ovarian growth of female Aedes albopictus by modulating the formation of JH receptor complex, expression of JH-inducible genes, and thereby disrupting JH-based endocrine regulations. These results suggested that farnesol and farnesyl acetate could be applicable for investigating underlying mechanisms of JH-regulated insect physiologies as well as developing novel eco-friendly insecticides.     

Fenoxycarb,a potent contaminant of the silkworm,Bombyx mori L., does not influence its juvenile hormone titer

Fenoxycarb (FC) effects were studied on juvenile hormone (JH) titers and JH‐esterase activities in the silkworm, B. Mori. In the literature, FC was observed to induce high JH titers but also to act without corpora allata (CA). These contradictory results did not permit us to conclude whether FC was a potent JH mimic or it was acting through the enhancement of JH titers in the hemolymph. Analysis of hemolymph JH‐esterase activities during the last larval instar reveals that FC was not a JH‐esterase inhibitor. Considering JHs, only JH II was detected in the European hybrid 200×300. Furthermore, JH titer was exactly identical in control and FC‐treated larvae, i.e., it dropped during the first 2 days of the last larval instar and became undetectable after day 2. This result is important since it contradicts the generally admitted concept that FC was acting by increasing the titer of JH. On the contrary, it was found that, despite its non‐terpenoid chemical structure, FC might be a JH mimic. In addition, FC suspected contamination of mulberry leaves was analyzed from a physiological viewpoint. We observed that "contaminated" mulberry leaves‐fed larvae became permanent larvae through the inhibition of their prothoracic glands (PG) activity and without any modification of their JH titers, i.e., exactly as for FC‐treated larvae. This last point adds information concerning the suspected implication of FC in the induction of the non‐spinning syndrome. Arch. Insect Biochem. Physiol. 40:141–149, 1999. © 1999 Wiley‐Liss, Inc.     

Action of a juvenile hormone analogue on the activity of Periplaneta americana corpora allata in vitro and on juvenile hormone III levels in vivo

The effects of the juvenile hormone (JH) analogue fenoxycarb (ethyl[2-(4-phenoxyphenoxy)-ethyl]carbamate) on the activity of corpora allata (CA) from adult female Periplaneta americana have been investigated. The in vitro biosynthesis of JH III by isolated CA was inhibited by about 85% in the presence of a high concentration (1 × 10⁻⁴ M) of fenoxycarb. However, at lower concentrations (1 × 10⁻⁶ M and 1 × 10⁻⁸ M) no inhibition of JH biosynthesis was apparent. Topical treatment of adult female cockroaches with fenoxycarb (100 μg/insect) did not reduce the subsequent rate of JH III biosynthesis by CA in vitro. By contrast, the same treatment markedly reduced the titre of endogenous JH III in intact cockroaches. These results suggest that CA activity in adult female P. americana may be controlled by negative feedback, and that this system of control is dependent on the maintenance of contact between the CA and nervous or humoral factors in the intact insect. Alternatively, it is possible that treatment with fenoxycarb increases the rate at which endogenous JH is metabolized.     

NMR assignments of juvenile hormone binding protein in complex with JH III

A hemolymph juvenile hormone binding protein (JHBP) shuttles hydrophobic JH, a key hormone in regulation of the insect life cycle, from the site of the JH biosynthesis to the cells of target organs. We report complete NMR chemical shift assignments of Bombyx mori JHBP in the JH III-bound state.     

Juvenile hormone activity of substituted aryl 3,7-dimethyl-6-octenyl ethers in the stable fly and house fly

Tests of substituted aryl 3,7-dimethyl-6-octenyl ethers and their epoxidized analogues produced a juvenile hormone (JH) mimic that was active against both the stable fly, Stomoxys calcitrans, and the house fly, Musca domestica. The criterion of activity in the JH bioassays was the formation of pupal-adult intermediates within the puparia.     

Bioassays of compounds with potential juvenoid activity on Drosophila melanogaster: Juvenile hormone III, bisepoxide juvenile hormone III and methyl farnesoates

Metabolites of the 6,7,10,11 bisepoxide juvenile hormone III (JHB₃), and other potential juvenoids, were tested for juvenile hormone activity using early instar or early stage pupae of Drosophila melanogaster. Importantly, methyl farnesoates were tested as they might have JH-like activity on Dipteran juveniles. Larvae were exposed to compounds in medium, or the compounds were applied to white puparia. In the assays employed in the present study, there was no indication for JH activity associated with the metabolites of JHB₃. The activity of methyl farnesoate (MF) was higher than that of JH III and far greater than bisepoxide JH III. As opposed to the two endogenous juvenile hormones, methyl farnesoate has weak activity in the white puparial bioassay. When fluorinated forms of methyl farnesoate, which is unlikely to be converted to JH, were applied to Drosophila medium to which fly eggs were introduced, there was a high degree of larval mortality, but no evidence of subsequent mortality at the pupal stage. One possible explanation for the results is that methyl farnesoate is active as a hormone in larval stages, but has little activity at the pupal stage where only juvenile hormone has a major effect.     

昆虫保幼激素受体的研究与应用进展

保幼激素(juvenile hormone, JH)是昆虫内分泌系统中的关键激素之一,对昆虫生长发育、变态、繁殖起着重要的调控作用。近年来有关JH的分子作用机制取得了极大的进展,主要得益于JH受体的鉴定,大量研究表明JH可通过胞内受体和膜受体两个途径来发挥生理调控功能。本文将从JH胞内受体Met的发现及鉴定、Met转录活性的调控因素、Met功能研究进展,以及Met作为JH受体在JH激动剂及拮抗剂筛选中的应用等方面对JH胞内受体的研究进展进行重点阐述;同时综述了有关JH膜受体的信号通路以及膜受体与核受体的互作等方面的研究进展。     

保幼激素在昆虫中的分子作用机理

随着分子生物学技术的快速发展,对生态环境中各类生物的研究,包括对生物某些特定基因结构和功能的研究等逐步拓展和加深。保幼激素(Juvenile Hormone,JH)是由咽侧体(Corpus Allatum,CA)分泌的,在昆虫发育、变态和生殖过程中起重要作用的激素。目前对JH信号传导途径的作用机理还不十分清楚。现有研究表明,Kruppel homolog-1(Kr-h1)是一种含C2H2锌指结构的转录因子,处于保幼激素信号途径下游,在保幼激素信号通路中起着重要作用。已报道的Kr-h1基因的功能主要包括:调控幼虫生长发育和变态,与蜜蜂的觅食行为密切相关,参与果蝇幼体神经细胞的形成等等。对就近十年来Kr-h1基因的特性和功能研究作一个综述以了解不同昆虫中保幼激素的分子作用机制,为开发生物农药奠定理论基础,也为维护良好的生态环境作出理论贡献。     

Morphogenetic effects of juvenile hormone and juvenile hormone mimics on adult development of Drosophila

The morphogenetic effects of t,t-farnesol, Law-Williams juvenile hormone analogue, dichlorofarnesenic acid ethyl ester (DFAEE), and a syntetic racemic or isomeric mixture of C₁₈ juvenile hormone (JH), when applied topically to pharate pupae and adults of D. melanogaster have been studied. Of these various agents tested, only DFAEE and JH affected adult development and eclosion and the pharate pupae were the most sensitive to these agents. The racemic mixture of JH induced the secretion, in the abdomen, of a supernumerary cuticle indistinguishable from that of the pupa; it, in addition, retarded the synthesis of brown eye pigments, general body pigmentation, and affected the differentiation of various internal organs and cuticular structures of the abdomen. By comparing the effects of JH with those of Minute (M) and bobbed (bb) mutations on the adult development, it is suggested that JH, by retarding genetic translation mimics M or bb.     

The isolation and identification of juvenile hormone from cockroach corpora allata in vitro

Methyl (2E, 6E) - 10, 11 - epoxy - 3, 7, 11 - trimethyl - 2, 6 - dodecadienoate (JH III) is synthesized by corpora cardiaca/allata of the American cockroach $\text{Periplaneta americana in vitro}$ and released into the culture medium. It constitutes the principal part of the biologically detectable JH activity. A fully synthetic medium without terpenoid precursors or serum was used, which proves, that the entire molecule is synthesized $\text{de novo}$. The presence of JH III in orders as different as Blattodea, Orthoptera, Coleoptera and Lepidoptera indicates, that it is the most widely occuring JH of insects.     

Diapause of the southwestern corn borer, Diatraea grandiosella: Further evidence showing juvenile hormone to be the regulator

Further evidence is presented to demonstrate the involvement of juvenile hormone (JH) in regulating diapause in the final larval stage of the southwestern corn borer. Diatraea grandiosella. JH titres in the haemolymph were measured throughout the entire diapause period. Additional results showed that actively secreting corpora allata are necessary to maintain diapause because allatectomized larvae terminated diapause prematurely. A topical application of JH mimic 2 days after the allatectomy prevented this premature termination of diapause. Intact nervous connections between the brain and the corpora allata were necessary for the maintenance of JH secretion. Other surgical work showed that the brains of nondiapausing larvae exhibited a higher ecdysiotropic activity than those of pre-, early-or mid-diapausing larvae.A single application of a JH mimic was more effective in maintaining a diapause-like state in nondiapausing larvae than were repeated topical applications of C₁₈-JH or an implantation of active corpora allata, suggesting that JH was more rapidly metabolized than was the JH mimic. The oxygen consumption of diapausing larvae which had received repeated topical applications of JH mimic was not significantly elevated over that of the controls indicating that treated larvae maintained a low metabolic rate even though they reverted to the spotted morph. A single application of 0.03 μg JH mimic/larva was sufficient to prolong diapause, thereby confirming that JH is necessary for diapause maintenance.     

Mosquitocidal activity of penfluridol as juvenile hormone antagonist

Juvenile hormone antagonists (JHANs) are known to interfere with the formation of juvenile hormone (JH) receptor complex. JHANs might be effective for control of target pests in larval stages at which stages high level of endogenous JH titer is maintained. In order to identify novel insecticidal compounds, 2352 compounds were surveyed on their JHAN activities using the yeast-two hybrid system. Among 53 compounds with JHAN activities, penfluridol showed high level of insecticidal activity against larvae of Aedes albopictus. JHAN activity was increased in proportion to the concentration of penfluridol. Larvicidal activity of penfluridol was 1.3–2.0 folds higher than that of pyriproxyfen. These results suggested that penfluridol could be useful for control of mosquito larvae.