Juvenile hormone analog binding in Manduca epidermis

Of a series of derivatives of JH I and methoprene, iodovinylmethoprenol (IVMA) proved most active (ed₅₀s of 3.2 pmol and 20 nM in the Manduca black larval assay and in the prevention of the 20-hydroxyecdysone-induced change to pupal commitment in the epidermis in vitro, respectively). When incubated with nuclei isolated from day 1 fifth instar abdominal epidermis, [¹²⁵I]IVMA bound specifically to two components with K_ds of 4 and 59 nM. This binding was competed by IVMA and methoprene but not by JH I; similar binding by [³H]JH I was not competed by IVMA or methoprene. Specific binding of IVMA was not found in pupally committed epidermis from wandering larvae, but it reappeared in pupal abdominal and thoracic epidermis but not in the wings. Culture of day 2 fifth instar epidermis with 20-hydroxyecdysone to cause pupal commitment caused the loss of the IVMA nuclear binders whereas culture in the absence of hormortes had no effect, indicating that 20HE in the absence of JH downregulates JH receptors.     

Juvenile hormone and juvenile hormone mimics inhibit proliferation in a lepidopteran imaginal disc cell line

The action of juvenile hormone (JH) and JH mimics have been examined in vitro by utilizing the imaginal disc-derived cell line, IAL-PID2. We have discovered that the cell line was responsive to JH and a variety of JH mimics. The most consistent response obtained in our studies was inhibition of cell proliferation, in the absence of 20-hydroxyecdysone (20E), which characteristically reduces cell proliferation in its own right in this cell line. JH-I, JH-III, methoprene, fenoxycarb, and farnesol significantly inhibited cell proliferation after 3 days of exposure of the cells in vitro to each of the compounds. Linoleic acid controls had no effect on proliferation in the cultures. The cell proliferation assay demonstrates the JH responsiveness of this cell line, but the concentrations of JH required were high compared to the concentrations of 20E needed for inhibition of proliferation in these cells.     

Ionotropic glutamate receptors mediate juvenile hormone synthesis in the cockroach,Diploptera punctata

By monitoring changes in the cytosolic [Ca2+](i) and rates of juvenile hormone (JH) synthesis in response to L-glutamate agonists and antagonists, we identified and characterized glutamate receptor subtypes in corpus allatum (CA) cells of the cockroach, Diploptera punctata. During the first ovarian cycle, corpora allata exhibited a cycle of changes in sensitivity to L-glutamate correlated to cyclic changes in rates of JH synthesis. When exposed to 60 microM L-glutamate in vitro, the active corpora allata of day-4 mated females produced 60% more JH, while inactive corpora allata at other ages showed 10-20% stimulatory response. Pharmacological characterization using various L-glutamate receptor agonists and antagonists indicated that several ionotropic subtypes of L-glutamate receptors were present in the CA. The CA showed an increase in rates of JH synthesis in response to NMDA, kainate, and quisqualate, but not to AMPA in both L-15 medium and minimum incubation medium. In contrast, applications of the metabotropic receptor-specific agonist trans-ACPD failed to elicit a change in the cytosolic [Ca2+](i) and JH production.An elevation of cytosolic calcium concentration, followed by 20-30% rise in JH production, was observed when active CA cells were exposed to 10-40 microM kainate. Kainate had no stimulatory effect on JH synthesis in calcium-free medium. The kainate-induced JH synthesis was blocked by 20 microM CNQX but was not affected by 20 microM NBQX. Kainate-stimulated JH production was not suppressed by MK-801 (a specific blocker of NMDA-receptor channel), nor was NMDA-stimulated JH production affected by CNQX (a specific antagonist of kainate receptor). These data suggest that active CA cells are stimulated to synthesize more JH by a glutamate-induced calcium rise via NMDA-, kainate- and/or quisqualate-sensitive subtypes of ionotropic L-glutamate receptors. The metabotropic-subtype and ionotropic AMPA-subtype L-glutamate receptors are unlikely to be present on active CA cells.     

Hormonal Control of Epidermal Cell Development

SYNOPSIS. During larval life the insect epidermis makes a larvalcuticle and certain pigments due to the presence of juvenilehormone (JH) at critical times during the molt cycle. The presenceof JH also permits growth of imaginal discs and maintains strictlylarval epidermis. At metamorphosis the lepidopteran epidermisresponds to a low level of 20- hydroxyecdysone (20HE) in theabsence of JH by becoming pupally committed, then later it formsa pupal cuticle when more 20HE appears, even though JH is present.During the change of commitment, DNA synthesis occurs but isnot essential, whereas both RN A and protein synthesis are.The major changes in the translatable mRNA population at thistime are threefold: a decline in most larval cuticle mRNAs,a transient increase followed by a disappearance of a few larvalcuticle mRNAs, and an appearance of at least one ‘pupalcommitment’ mRNA and two to three mRNAs for small pupalcuticular proteins. Similar changes are seen in the proteinsynthetic patterns. Thus, a pupally committed cell is one whichcan no longer make larval products but which is not yet ableto make most pupal products. Juvenile hormone prevents the changeto pupal commitment by directing some of both the primary andthe secondary actions of 20HE on the genome.     

Interaction of hormones in controlling reproductive function of Drosophila females under stressful conditions is genetically determined

The effect of heat stress (38 degrees C) on the content of octopamine (OA) and 20-hydroxyecdysone (20HE) was studied under normal and stressful conditions in adult flies of Drosophila virilis lines contrasting in the level of the juvenile hormone (JH). The wild-type flies (line 101) exhibited a pronounced sex dimorphism for the content of both OA and 20HE, which was substantially lower in this line than in flies of the mutant line 147. The level of both hormones increased in flies of line 101 exposed to heat stress, whereas it remained unchanged in flies of line 147 under the same conditions. The effect of heat stress on the level of JH metabolism and fertility was also studied in D. melanogaster wild-type lines and lines carrying mutations in genes responsible for OA and DA syntheses. In octopamineless females of the T beta hnM18 line and in females of the Ste line characterized by a doubled content of DA, JH degradation differed from normal: it was increased in both young and mature T beta hnM18 females, while decreased in young and increased in mature Ste flies. Fertility was substantially lower in the Ste than in the wild-type line. Flies of all of the D. melanogaster lines produced a stress response; however, in mutant lines, both fertility and stress reactivity of the systems controlling JH metabolism differed significantly from that of the wild-type lines. The role of JH, 20HE, OA, and DA interaction in regulation of Drosophila reproduction under stressful conditions is discussed.     

外源保幼激素对中华通草蛉滞育解除及滞育后发育的影响

[目的]本研究旨在明确外源保幼激素(juvenile hormone,JH)在中华通草蛉Chrysoperla sinica滞育快速解除过程中的使用剂量及最佳使用时期.[方法]测定点滴不同剂量(0,5,15,25和35 μg/成虫)外源JH后中华通草蛉滞育成虫的产卵前期、产卵历期、雌虫寿命及单雌产卵量,以及15 μg/...     

Reduced titres of ecdysone following juvenile hormone treatment in the German cockroach, Blattella germanica

Changes in ecdysone titre of the larvae of the German cockroach, Blattella germanica, exposed continuously to the juvenile hormone (JH), or to the insect growth regulator (IGR) with JH activity, can be correlated with the nature of the substance applied, its dose, and the time of application. The younger larvae exposed to the high dose of the IGR die in the next ecdysis, whereas the same treatment induces a diapause-like stage of developmental arrest in the last larval stage. The affected larvae have very little or no ecdysone, the synthesis of which takes place in the second part of the instar. The same treatment after this period has a lesser effect. The extent of the effect is correlated to the amount of ecdysone synthesized before the application of IGR. Last instar larvae exposed to the lower dose of the IGR or JH lack the peak of ecdysone normally found in the controls at the end of the second third of the instar when metamorphosis takes place. In these insects the first rise of the ecdysone titre begins towards the end of the instar, and ecdysis into the supernumerary larval stage is initiated when the ecdysone titre reached a level permitting ecdysis.A direct or indirect antagonism between these hormones, both fundamental to insect development, can explain the morphogenetic, inhibitory, and lethal effects observed in insects treated with JH or IGR with JH activity.     

Modulation of juvenile hormone release and oocyte growth following (RS)-hydroprene treatment in virgin female Diploptera punctata

ABSTRACT. The effect of (flS)-hydroprene treatment (2, 20, 200 μ g) on JH release was assessed in virgin females of D. punctata (Eschscholtz) during the first 10 days of adult life as was basal oocyte length and number of cells in the CA. At a dose of 2 μ g hydroprene, JH release was stimulated slightly and, on days 4 and 6, oocyte growth was significantly greater than that of acetone-treated controls. A similar but more striking enhancement of JH release and basal oocyte growth was observed at a dose of 20 μ g and a significant inhibition of JH release, in concert with a rapid growth of basal oocytes, was observed at a dose of 200 μ g. During the observation period, the mean number of cells in the CA decreased in a dose-dependent fashion, with a highly significant reduction in numbers in 20 and 200 μ g-treated animals. Reimplantation of vitellogenic ovarioles (three or six) into ovariectomized virgin females also resulted in an enhancement of JH release; this indicates that virgin female CA can respond to the stimulatory action of the ovary and is consistent with a model for ( RS )-hydroprene action in which the 'positive feedback' effect (stimulation of JH release) observed with low doses of the analogue occurs as a consequence of the action of the analogue on the ovary. ( RS )-hydroprene treatment stimulates basal oocyte growth to the point at which the previously unstimulatory virgin oocytes are able to enhance JH release by a feedback loop involving the CA and probably the brain.     

Temporal patterns of protein synthesis inManduca epidermis during the change to pupal commitment in vitro: their modulation by 20-hydroxyecdysone and juvenile hormone

Summary The epidermis of final instar tobacco hornworm larvae,Manduca sexta, becomes committed to pupal differentiation in response to ecdysteroid in the absence of juvenile hormone (JH). Many changes in protein synthetic patterns have been noted during this time (Kiely and Riddiford 1985). To determine which of these changes are caused by ecdysteroid and which are important for the change of commitment, we have incubated larvally-committed epidermis for 24 h with 1 g/ml 20-hydroxyecdysone (20HE) and 3 g/ml epoxygeranylsesamole (EGS) (a JH mimic), with 3 g/ml EGS alone, or in hormone-free medium. Synthesis of larval-specific proteins such as insecticyanin and larval cuticular proteins was reduced to trace amounts or was undetectable after culture with 20HE for 24 h. The larval cuticular proteins that are greatly increasedin vivo on day 3 were not synthesized after exposure to 20HEin vitro. Ecdysteroid increased the synthesis of many of the proteins first seenin vivo on day 3 or during the wandering stage. The synthesis of about half of these latter proteins was inhibited by JH, indicating that they were likely part of the change of commitment. Other proteins that appear at this stagein vivo showed increased synthesis also in hormone-free medium and therefore were independent of the change of commitment.     

Antagonistic effect of juvenile hormone on hemocyte-spreading behavior of Spodoptera exigua in response to an insect cytokine and its putative membrane action

Juvenile hormone (JH) acts on membrane of follicle cells to induce ovarian patency for vitellogenesis, though it regulates various other physiological processes via putative intracellular receptors. This study suggests another JH membrane action by analyzing in vitro hemocyte behavior. In response to nonself, both granular cells and plasmatocytes of Spodoptera exigua can exhibit cell shape changes through spreading behaviors. Plasmatocytes were separated from total S. exigua hemocytes by Percoll gradient and exposed in vitro to an insect cytokine, plasmatocyte-spreading peptide (PSP), identified from Pseudoplusia includens. In response, the purified plasmatocytes spread in a dose-dependent manner from picomolar to micromolar concentrations. Interestingly, the PSP responses of plasmatocytes in S. exigua varied among different larval ages during fifth instar ( approximately 5 days at 25 degrees C) in a sensitivity order of late (5 days old)相似文献   

The participation of calponin in the cross talk between 20-hydroxyecdysone and juvenile hormone signaling pathways by phosphorylation variation

20-hydroxyecdysone (20E) and juvenile hormone (JH) signaling pathways interact to mediate insect development, but the mechanism of this interaction is poorly understood. Here, a calponin homologue domain (Chd) containing protein (HaCal) is reported to play a key role in the cross talk between 20E and JH signaling by varying its phosphorylation. Chd is known as an actin binding domain present in many proteins including some signaling proteins. Using an epidermal cell line (HaEpi), HaCal was found to be up-regulated by either 20E or the JH analog methoprene (JHA). 20E induced rapid phosphorylation of HaCal whereas no phosphorylation occurred with JHA. HaCal could be quickly translocated into the nuclei through 20E or JH signaling but interacted with USP1 only under the mediation of JHA. Knockdown of HaCal by RNAi blocked the 20E inducibility of USP1, PKC and HR3, and also blocked the JHA inducibility of USP1, PKC and JHi. After gene silencing of HaCal by ingestion of dsHaCal expressed by Escherichia coli, the larval development was arrested and the gene expression of USP1, PKC, HR3 and JHi were blocked. These composite data suggest that HaCal plays roles in hormonal signaling by quickly transferring into nucleus to function as a phosphorylated form in the 20E pathway and as a non-phosphorylated form interacting with USP1 in the JH pathway to facilitate 20E or JH signaling cascade, in short, by switching its phosphorylation status to regulate insect development.     

The ecdysoneless1 gene regulates metabolism of the juvenile hormone and dopamine in Drosophila melanogaster

The dopamine (DA) content and the level of juvenile hormone (JH) degradation were studied in females of the wild-type Canton S strain and the ecdysoneless1 (ecd1) mutant, which does not produce ecdysone at a restrictive temperature (29 degrees C). Exposure at the restrictive temperature considerably increased the JH-hydrolyzing activity and the DA content in five-day ecd1 females compared with flies of both strains growing at 19 degrees C and Canton S females exposed at 29 degrees C. In one-day ecd1 females, the level of JH degradation also increased at the restrictive temperature, but the DA content was low. The effect of ecdysone deficiency on the stress response in Drosophila melanogaster females was studied using changes in DA content and JH degradation were used as indices. The ecd1 mutation did not prevent the initiation of the stress response in females exposed at the restrictive temperature, but changed its intensity (stress reactivity). The interaction of 20-hydroxyecdysone with JH and DA in regulating Drosophila reproduction under normal conditions and in stress is discussed.     

The all-or-none rule in morphogenetic action of juvenile hormone on insect epidermal cells

We have investigated ultrastructural changes in the integuments of larval–adult and larval–pupal intermediates produced by exogenous application of juvenile hormone (JH) analogues in Pyrrhocoris apterus (Hemiptera), and Galleria mellonella and Manduca sexta (Lepidoptera). Ultrastructural analysis of the epidermis of these intermediates always revealed the presence of only two types of epidermal cell, which produced morphologically perfect cuticles of the previous and future developmental stages. There were no intermediate cuticles at the level of individual cells. It has been determined that a single epidermal cell constitutes the lowest elementary unit in the perception and realization of the developmental messages conveyed by JH to its target tissues. Further investigations revealed that the responses of individual epidermal cells to JH were strictly autonomous and qualitative, i.e. they were executed according to the ''yes-or-no'' or ''all-or-none'' rule. The neighbouring epidermal cells could realize independently, side-by-side, the quite dissimilar +JH (somatic growth) or -JH (metamorphosis) developmental programmes, although each of them formed biochemically, functionally, and ontogenetically different structures. The qualitative on- and off- signal given by JH for induction of the stationary (+JH) developmental cycle was limited to relatively short, genetically determined, and stage-specific developmental periods of cellular susceptibility to JH. The mosaic mixtures of the heterochronic, larval–pupal or adult epidermal cells, which we found in different proportions on the bodies of the intermediates, revealed two variable, development-related factors: (i) the presence or absence of a minimum effective concentration of JH, and (ii) positive or negative sensitivity of a particular epidermal cell to JH.     

Control of ommochrome synthesis by both juvenile hormone and melanization hormone in the cabbage armyworm,Mamestra brassicae

Summary Pigmentation of last instar larvae of the cabbage armyworm,Mamestra brassicae is of two types: melanin in the cuticle and ommochrome in the epidermis. The latter was found to be primarily xanthommatin. When allatectomy was performed 8 h before head capsule slippage (HCS) in the last larval molt, later ommochrome synthesis was inhibited. Application of juvenile hormone (JH) up to 12 h after HCS (9 h before ecdysis) (activity: methopreneJH I>JH II>JH III) restored ommochrome synthesis. After that time it has less and less effect.Removal of the suboesophageal ganglion from the larvae 8 h before HCS prevented both later ommochrome synthesis and melanization. Melanization of isolated abdomens was restored by implantation of 3 suboesophageal ganglia or injection of melanization and reddish coloration hormone (MRCH) 18 h after HCS. Restoration of ommochrome synthesis required exogenous JH in addition to melanization hormone from suboesophageal ganglion or MRCH. Therefore, melanization appears to be critical for the later onset of ommochrome synthesis even in a larva which has been exposed to JH during the critical period.Abbreviations CC·CA corpora cardiaca-corpora allata complex - JH juvenile hormone - MRCH melanization and reddish coloration hormone - HCS head capsule slippage     

In vitro metabolism of juvenile hormone III and juvenile hormone III bisepoxide by Drosophila melanogaster and mammalian cytosolic epoxide hydrolase

In vitro metabolism of juvenile hormone III (JH III) and juvenile hormone III bisepoxide was investigated using purified mouse liver cytosolic epoxide hydrolase (cEH) and cell fractions from Drosophila melanogaster. JH III was metabolized faster than JH III bisepoxide by epoxide hydrolase activity in D. melanogaster cell fractions and by cEH. After incubation with JH III bisepoxide, all cell fractions and cEH produced epoxy-diol, cis- and trans-tetrahydrofuran-diols, and tetraol as metabolites. An increase in the concentration of cEH resulted in an increase in the proportion of tetraol as a JH III bisepoxide metabolite but this trend was not observed in the D. melanogaster cell fractions. Differences between cell fractions in the metabolism of JH III and JH III bisepoxide suggests the presence of juvenile hormone epoxide hydrolase isozymes.     

Regulation of Reproductive Function in DrosophilaFemales Due to Hormonal Interaction Under Stress Is Genetically Determined

The effect of heat stress (38°C) on the content of octopamine (OA) and 20-hydroxyecdysone (20HE) was studied under normal and stressful conditions in adult flies of Drosophila virilislines contrasting in the level of the juvenile hormone (JH). The wild-type flies (line 101) exhibited a pronounced sex dimorphism for the content of both OA and 20HE, which was substantially lower in this line than in flies of the mutant line 147. The level of both hormones increased in flies of line 101 exposed to heat stress, whereas it remained unchanged in flies of line 147 under the same conditions. The effect of heat stress on the level of JH metabolism and fertility was also studied in D. melanogasterwild-type lines and lines carrying mutations in genes responsible for OA and DA syntheses. In octopamineless females of the Th ^nM18line and in females of the Steline characterized by a doubled content of DA, JH degradation differed from normal: it was increased in both young and mature Th ^nM18females, while decreased in young and increased in mature Steflies. Fertility was substantially lower in the Stethan in the wild-type line. Flies of all of the D. melanogasterlines produced a stress response; however, in mutant lines, both fertility and stress reactivity of the systems controlling JH metabolism differed significantly from that of the wild-type lines. The role of JH, 20HE, OA, and DA interaction in regulation of Drosophilareproduction under stressful conditions is discussed.     

Cell size control by ovarian factors regulates juvenile hormone synthesis in corpora allata of the cockroach, Diploptera punctata

The corpora allata synthesize and release juvenile hormone (JH) that in turn regulates insect growth, metamorphosis and reproduction. In the corpus allatum (CA) of the female adult cockroach Diploptera punctata, cyclic rise and decline in JH synthesis rates occur concurrently with cyclic growth and atrophy during an ovarian cycle. Here, we report that protein content decreases, whereas Golgi population, lysosomal content and autophagic activities increase with decrease in CA cell size. Also, the concentration of cyclic GMP (cGMP) is low in large cells and high in small cells. Results of treating CA with ovarian tissue suggest that a putative peptidergic growth regulator released from mature ovaries acts directly on active CA cells and induces the elevation of intracellular cGMP content. Consequently, elevated cGMP may inhibit protein synthesis or trigger massive and synchronous autophagic activities, resulting in cell atrophy and reduction of protein content. As a result of the depletion of cellular machinery, CA glands exhibit long-term depression in JH synthesis.