To gain insights into how juvenile hormone (JH) came to regulate insect metamorphosis, we studied its function in the ametabolous firebrat, Thermobia domestica. Highest levels of JH occur during late embryogenesis, with only low levels thereafter. Loss-of-function and gain-of-function experiments show that JH acts on embryonic tissues to suppress morphogenesis and cell determination and to promote their terminal differentiation. Similar embryonic actions of JH on hemimetabolous insects with short germ band embryos indicate that JH's embryonic role preceded its derived function as the postembryonic regulator of metamorphosis. The postembryonic expansion of JH function likely followed the evolution of flight. Archaic flying insects were considered to lack metamorphosis because tiny, movable wings were evident on the thoraces of young juveniles and their positive allometric growth eventually allowed them to support flight in late juveniles. Like in Thermobia, we assume that these juveniles lacked JH. However, a postembryonic reappearance of JH during wing morphogenesis in the young juvenile likely redirected wing development to make a wing pad rather than a wing. Maintenance of JH then allowed wing pad growth and its disappearance in the mature juvenile then allowed wing differentiation. Subsequent modification of JH action for hemi- and holometabolous lifestyles are discussed.

• Keywords
• developmental biology, differentiation, ecdysone, juvenile hormone, metamorphosis, myoglianin, precocene, thermobia domestica,
• MeSH
• Metamorphosis, Biological * physiology   MeSH
• Insecta MeSH
• Juvenile Hormones * MeSH
• Morphogenesis MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Juvenile Hormones * MeSH

Germ cells are essential to sexual reproduction. Across the animal kingdom, extracellular signaling isoprenoids, such as retinoic acids (RAs) in vertebrates and juvenile hormones (JHs) in invertebrates, facilitate multiple processes in reproduction. Here we investigated the role of these potent signaling molecules in embryonic germ cell development, using JHs in Drosophila melanogaster as a model system. In contrast to their established endocrine roles during larval and adult germline development, we found that JH signaling acts locally during embryonic development. Using an in vivo biosensor, we observed active JH signaling first within and near primordial germ cells (PGCs) as they migrate to the developing gonad. Through in vivo and in vitro assays, we determined that JHs are both necessary and sufficient for PGC migration. Analysis into the mechanisms of this newly uncovered paracrine JH function revealed that PGC migration was compromised when JHs were decreased or increased, suggesting that specific titers or spatiotemporal JH dynamics are required for robust PGC colonization of the gonad. Compromised PGC migration can impair fertility and cause germ cell tumors in many species, including humans. In mammals, retinoids have many roles in development and reproduction. We found that like JHs in Drosophila, RA was sufficient to impact mouse PGC migration in vitro. Together, our study reveals a previously unanticipated role of isoprenoids as local effectors of pre-gonadal PGC development and suggests a broadly shared mechanism in PGC migration.

• Keywords
• Hmgcr, cell movement, embryonic development, gametogenesis, germ cells, gonad, juvenile hormones, ovary, retinoids, testis,
• MeSH
• Drosophila melanogaster * MeSH
• Drosophila MeSH
• Gonads MeSH
• Juvenile Hormones * MeSH
• Humans MeSH
• Mice MeSH
• Cell Movement MeSH
• Mammals MeSH
• Terpenes MeSH
• Germ Cells MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Names of Substances
• Juvenile Hormones * MeSH
• Terpenes MeSH

To gain insights into how juvenile hormone (JH) came to regulate insect metamorphosis, we studied its function in the ametabolous firebrat, Thermobia domestica. Highest levels of JH occur during late embryogenesis, with only low levels thereafter. Loss-of-function and gain-of-function experiments show that JH acts on embryonic tissues to suppress morphogenesis and cell determination and to promote their terminal differentiation. Similar embryonic actions of JH on hemimetabolous insects with short germ band embryos indicate that JH's embryonic role preceded its derived function as the postembryonic regulator of metamorphosis. The postembryonic expansion of JH function likely followed the evolution of flight. Archaic flying insects were considered to lack metamorphosis because tiny, movable wings were evident on the thoraces of young juveniles and their positive allometric growth eventually allowed them to support flight in late juveniles. Like in Thermobia, we assume that these juveniles lacked JH. However, a postembryonic reappearance of JH during wing morphogenesis in the young juvenile likely redirected wing development to make a wing pad rather than a wing. Maintenance of JH then allowed wing pad growth and its disappearance in the mature juvenile then allowed wing differentiation. Subsequent modification of JH action for hemi- and holometabolous lifestyles are discussed.

• Keywords
• differentiation, ecdysone, juvenile hormone, metamorphosis, myoglianin, precocene,
• Publication type
• Journal Article MeSH
• Preprint MeSH

Insect Growth Regulators (IGRs) represent advanced, bio-rational insecticides. This Special Issue reflects progress in IGR development that has been enabled by insight into the molecular principles of biosynthetic or hormone signaling pathways. The unifying principle is aiming at processes and molecular targets that are unique to arthropods and ideally to narrower insect taxa representing pests or disease vectors. While some strategies of obtaining the desired compounds for chemical intervention rely on rational, structure-based design or computational power, others exploit technologies allowing automated, high-throughput screening of large chemical libraries. All avenues leading to selective and environmentally safe pest control are valid as we face the imminent threat of the declining world insect population.

• Keywords
• biosynthesis, ecdysone, juvenile hormone, metamorphosis, molting, receptors,
• Publication type
• Journal Article MeSH

Insect metamorphosis boasts spectacular cases of postembryonic development when juveniles undergo massive morphogenesis before attaining the adult form and function; in moths or flies the larvae do not even remotely resemble their adult parents. A selective advantage of complete metamorphosis (holometaboly) is that within one species the two forms with different lifestyles can exploit diverse habitats. It was the environmental adaptation and specialization of larvae, primarily the delay and internalization of wing development, that eventually required an intermediate stage that we call a pupa. It is a long-held and parsimonious hypothesis that the holometabolous pupa evolved through modification of a final juvenile stage of an ancestor developing through incomplete metamorphosis (hemimetaboly). Alternative hypotheses see the pupa as an equivalent of all hemimetabolous moulting cycles (instars) collapsed into one, and consider any preceding holometabolous larval instars free-living embryos stalled in development. Discoveries on juvenile hormone signalling that controls metamorphosis grant new support to the former hypothesis deriving the pupa from a final pre-adult stage. The timing of expression of genes that repress and promote adult development downstream of hormonal signals supports homology between postembryonic stages of hemimetabolous and holometabolous insects. This article is part of the theme issue 'The evolution of complete metamorphosis'.

• Keywords
• evolution, hormone receptor, juvenile hormone, metamorphosis, signal transduction, transcription factor,
• MeSH
• Metamorphosis, Biological * MeSH
• Insecta classification   growth & development   MeSH
• Juvenile Hormones metabolism   MeSH
• Pupa growth & development   MeSH
• Signal Transduction * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Juvenile Hormones MeSH

The sesquiterpenoid juvenile hormone (JH) is vital to insect development and reproduction. Intracellular JH receptors have recently been established as basic helix-loop-helix transcription factor (bHLH)/PAS proteins in Drosophila melanogaster known as germ cell-expressed (Gce) and its duplicate paralog, methoprene-tolerant (Met). Upon binding JH, Gce/Met activates its target genes. Insects possess multiple native JH homologs whose molecular activities remain unexplored, and diverse synthetic compounds including insecticides exert JH-like effects. How the JH receptor recognizes its ligands is unknown. To determine which structural features define an active JH receptor agonist, we tested several native JHs and their nonnative geometric and optical isomers for the ability to bind the Drosophila JH receptor Gce, to induce Gce-dependent transcription, and to affect the development of the fly. Our results revealed high ligand stereoselectivity of the receptor. The geometry of the JH skeleton, dictated by two stereogenic double bonds, was the most critical feature followed by the presence of an epoxide moiety at a terminal position. The optical isomerism at carbon C11 proved less important even though Gce preferentially bound a natural JH enantiomer. The results of receptor-ligand-binding and cell-based gene activation assays tightly correlated with the ability of different geometric JH isomers to induce gene expression and morphogenetic effects in the developing insects. Molecular modeling supported the requirement for the proper double-bond geometry of JH, which appears to be its major selective mechanism. The strict stereoselectivity of Gce toward the natural hormone contrasts with the high potency of synthetic Gce agonists of disparate chemistries.

• Keywords
• Drosophila, basic helix-loop-helix transcription factor (bHLH), development, hormone receptor, insect, juvenile hormone (JH), ligand-binding protein, reproduction, stereoselectivity,
• MeSH
• Drosophila melanogaster chemistry   genetics   metabolism   MeSH
• Juvenile Hormones chemistry   metabolism   MeSH
• Models, Molecular MeSH
• Drosophila Proteins metabolism   MeSH
• Receptors, Cell Surface metabolism   MeSH
• Stereoisomerism MeSH
• Basic Helix-Loop-Helix Transcription Factors metabolism   MeSH
• Transcription Factors metabolism   MeSH
• Protein Binding MeSH
• Gene Expression Regulation, Developmental MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• gce protein, Drosophila MeSH Browser
• Juvenile Hormones MeSH
• MET protein, Drosophila MeSH Browser
• Drosophila Proteins MeSH
• Receptors, Cell Surface MeSH
• Basic Helix-Loop-Helix Transcription Factors MeSH
• Transcription Factors MeSH

Juvenile hormones (JH) and ecdysteroids regulate many biological and metabolic processes. CREB-binding protein (CBP) is a transcriptional co-regulator with histone acetyltransferase (HAT) activity. Therefore, CBP is involved in activation of many transcription factors that regulate expression of genes associated with postembryonic development in insects. However, the function of CBP in JH action in insects is not well understood. Hence, we studied the role of CBP in JH action in the red flour beetle, Tribolium castaneum and the Tribolium cell line. CBP knockdown caused a decrease in JH induction of genes, Kr-h1, 4EBP and G13402 in T. castaneum larvae, adults and TcA cells whereas, Trichostatin A [TSA, a histone deacetylase (HDAC) inhibitor] induced the expression of these JH-response genes. Western blot analysis with specific antibodies revealed the requirement of CBP for the acetylation of H3K18 and H3K27 in both T. castaneum and TcA cells. Chromatin immunoprecipitation (Chip) assays showed the importance of CBP-mediated acetylation of H3K27 for JH induction of Kr-h1, 4EBP, and G13402 in TcA cells. These data suggest that CBP plays an important role in JH action in the model insect, T.castaneum.

• MeSH
• Acetylation MeSH
• Gene Knockout Techniques MeSH
• Histones metabolism   MeSH
• Insect Proteins genetics   metabolism   MeSH
• Juvenile Hormones pharmacology   MeSH
• CREB-Binding Protein genetics   metabolism   MeSH
• Tribolium genetics   growth & development   metabolism   MeSH
• Gene Expression Regulation, Developmental drug effects   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Names of Substances
• Histones MeSH
• Insect Proteins MeSH
• Juvenile Hormones MeSH
• CREB-Binding Protein MeSH

BACKGROUND: Juvenile hormones (JH) and ecdysteroids control postembryonic development in insects. They serve as valuable targets for pest management. Hence, understanding the molecular mechanisms of their action is of crucial importance. CREB-binding protein (CBP) is a universal transcriptional co-regulator. It controls the expression of several genes including those from hormone signaling pathways through co-activation of many transcription factors. However, the role of CBP during postembryonic development in insects is not well understood. Therefore, we have studied the role of CBP in postembryonic development in Tribolium, a model coleopteran insect. RESULTS: CBP is ubiquitously expressed in the red flour beetle, Tribolium castaneum. RNA interference (RNAi) mediated knockdown of CBP resulted in a decrease in JH induction of Kr-h1 gene expression in Tribolium larvae and led to a block in their development. Moreover, the injection of CBP double-stranded RNA (dsRNA) showed lethal phenotypes within 8 days of injection. RNA-seq and subsequent differential gene expression analysis identified CBP target genes in Tribolium. Knockdown of CBP caused a decrease in the expression of 1306 genes coding for transcription factors and other proteins associated with growth and development. Depletion of CBP impaired the expression of several JH response genes (e.g., Kr-h1, Hairy, early trypsin) and ecdysone response genes (EcR, E74, E75, and broad complex). Further, GO enrichment analyses of the downregulated genes showed enrichment in different functions including developmental processes, pigmentation, anatomical structure development, regulation of biological and cellular processes, etc. CONCLUSION: These data suggest diverse but crucial roles for CBP during postembryonic development in the coleopteran model insect, Tribolium. It can serve as a target for RNAi mediated pest management of this stored product pest.

• Keywords
• CBP, Ecdysone, Juvenile hormone, Kr-h1, RNA seq, RNAi, Tribolium,
• MeSH
• Gene Expression MeSH
• Insect Proteins antagonists & inhibitors   genetics   metabolism   physiology   MeSH
• Juvenile Hormones pharmacology   MeSH
• Larva genetics   metabolism   MeSH
• CREB-Binding Protein antagonists & inhibitors   genetics   metabolism   physiology   MeSH
• RNA Interference MeSH
• Tribolium genetics   growth & development   metabolism   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Names of Substances
• Insect Proteins MeSH
• Juvenile Hormones MeSH
• CREB-Binding Protein MeSH

The physiological and biochemical mechanisms that enable insects to feed on dry food to secure enough water for larval growth were investigated. The study was carried out with a plethora of physiological methods, ranging from the simple volumetric determination of O2 consumption and water intake to more advanced methods such as scanning microrespirography and thermovision imaging of insect's body temperature. The experiments were done on the European firebug, Pyrrhocoris apterus, which feeds exclusively on dry linden seeds. In order to survive, it needs to drink water or suck a sap from plants occasionally. It was found that the young larval instars compensate the occasional water deficiency by the increased production of metabolic water. The juvenile hormone (JH)-dependent production of metabolic water, which was previously found in other species consuming dry food, was achieved in P. apterus by total metabolic combustion of the dietary lipid (neutral seed oil). The water-producing, hypermetabolic larvae were heated from inside by endothermic energy released from the uncoupling of oxidation from oxidative phosphorylation. The "warm", hypermetabolic larvae burning the dietary oil into CO2 and water showed the increased rates of respiratory metabolism. Microrespirographic recording of these larvae revealed the ratio of the respiratory quotient (RQ, CO2/O2) of 0.7, which indicated the breakdown of a pure triglyceride. The warm hypermetabolic larvae could be easily spotted and distinguished from the "cold" larvae on the screen of a thermovision camera. The last instar larvae lacking the JH were always only cold. They metabolized a carbohydrate substrate exclusively (RQ = 1.0), while the dietary lipid was stored in the fat body. In comparison with the hypermetabolic larvae of some other species fed on dry food, which exhibited the highest rates of O2 consumption ever recorded in a living organism (10-20 mL O2/g per hour), the metabolic difference between the warm and cold larvae of P. apterus was only some 30% (not a reported 10-fold difference), which was presumably due to their ability to drink. We conclude that a very important, though still largely neglected, epigenetic biochemical role of insect JH depends on switchover between the utilization of dietary lipid (+JH; production of metabolic water) and carbohydrate (-JH; lipid storage in the fat body). The hypermetabolic water supply in insects fed on dry food, which is associated with enormous rates of O2 consumption, liberates endothermic energy that heats the body and potentially influences the insect thermoregulation. A possibility that the JH-dependent lipolytic hormone stimulates the total metabolic breakdown of nutritional lipids may be absolutely different from the currently known adipokinetic peptides that have been emphasized.

• Keywords
• JH, O2 consumption, O2), Pyrrhocoris apterus, endothermic energy, hypermetabolism, juvenile hormone, respiratory quotient (CO2, uncoupling of oxidation, “warm” and “cold” larvae,
• Publication type
• Journal Article MeSH

• MeSH
• Aedes genetics   MeSH
• Genes, Insect * MeSH
• Insect Proteins metabolism   MeSH
• Juvenile Hormones metabolism   MeSH
• Methoprene metabolism   MeSH
• Gene Expression Regulation * MeSH
• Repressor Proteins metabolism   MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comment MeSH
• Names of Substances
• Insect Proteins MeSH
• Juvenile Hormones MeSH
• Methoprene MeSH
• Repressor Proteins MeSH