Possible involvement of ecdysteroids in photoperiodically induced suppresion of ovarian development in a Japanese strain of the migratory locust,Locusta migratoria

In a Japanese population of Locusta migratoria, adult females become reproductively inactive under crowding and long days (LD) and reproductively active under crowding and short days (SD). The identity and titre of ecdysteroids in the haemolymph and ovaries from adult females reared under SD and LD were investigated by RIA/HPLC. The effects of exogenous juvenile hormone (JH) III treatments on the termination of such reproductive arrest and ecdysteroid contents in LD females were also examined. In general, ecdysteroid titres in both haemolymph and ovaries were significantly higher in reproductively active SD females than in reproductively inactive LD females. A clear difference was also observed in oocyte growth between SD and LD individuals. JH III applications (three consecutive topical applications, 150 μg per insect per day from day 3) stimulated ovarian development in LD females and significantly increased the haemolymph and ovarian ecdysteroids to a level comparable to that of reproductively active SD adult females.     

Juvenile hormone regulation on the flight capability of Bactrocera dorsalis (Diptera: Tephritidae)

The oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is considered a major economic threat in many regions worldwide. In order to better understand the flight capacity of B. dorsalis and its physiological basis, the functions and regulatory roles of juvenile hormone (JH) in the flight muscle of B. dorsalis were studied under a controlled environment. JH titer of B. dorsalis varied with age and sex. Females have higher JH titers and better flight capabilities than males, given that the increase in JH also corresponded to the ovarian development and maturation in females. The flight duration and distance of both males and females increased with the gradual increase of JH titer after adult emergences. Both JH titer and flight capability peaked in 15-d-old adult and declined subsequently with age. Flight activity stimulated the production of JH as adults flown for 24 h on the flight mills have the highest JH titers compare to adults flown on shorter flight durations. Adults treated with 0.5 µg and 5 µg of JH III were able to fly long durations and long distances, nevertheless when JH titer was too low or too high, it would restrict the flight ability of the fly. The mutual reinforcement of JH and flight activity provides fundamental understanding on the physiological aspects of the flight capability and dispersal, which facilitates strategies for the long-term control of this destructive pest.     

Structure,haemolymph titre,and regulatory effects of juvenile homone during ovarian maturation in Leucophaea maderae

Juvenile hormone (JH) synthesized and secreted in vitro by the corpora allata of mated adult Leucophaea maderae females was determined to be JH III (methyl-10,11-epoxy-3,7,11-trimethyl-2,6-dodecadienoate).The haemolymph titre of JH was determined during maturation of the terminal oöcytes in the first reproductive cycle of L. maderae. In virgin females, JH is not detectable in the haemolymph during the first eight days following adult emergence; however, by 10 days after emergence, trace quantities of JH are apparent. Mating stimuli induce a dramatic increase in the concentration of haemolymph JH, with a peak occurring approximately 12 days after mating; thereafter, the JH concentration declines until it has reached an undetectable level 19 days after mating, at the time of chorion deposition.During ovarian maturation, changes in the rates of synthesis of vitellogenin by the fat body and DNA by the ovary correlate closely with the haemolymph titre of JH. However, no such correlation exists between the JH titre and the extensive ovarian protein synthesis that occurs in L. maderae coincident with chorion formation.The effects of JH I and JH III on both vitellogenin synthesis and ovarain DNA synthesis are statistically similar.     

Activity of the corpora allata in the adult female migratory locust

Juvenile hormone was detected in the haemolymph of adult female Locusta by a modified Galleria bioassay. The hormone was present in the haemolymph immediately after the final ecdysis, but could not be detected after this time until the end of the period of somatic growth just before the start of ovarian development. During the first gonotrophic cycle the levels of juvenile hormone in the haemolymph could be related to the growth of the proximal oöcytes. The volumes of the corpora allata could be related to haemolymph juvenile hormone levels during the first gonotrophic cycle. Ovariectomy had no effect on haemolymph juvenile hormone levels or on the volumes of the corpora allata.     

Detection of juvenile hormone I,II and III in adult monarch butterflies (Danaus plexippus)

Hemolymph of Danaus plexippus (monarch butterfly) was analyzed for juvenile hormone titer by gas liquid chromatography-mass spectrometry (GC-MS) in the selected ion monitoring mode. Laboratory reared, reproductively active, adult males and females contained JH I, II and III. JH II predominated with titers ranging from 2.3 to 11 ng/ml hemolymph. Titers of JH I and JH III were approx. 1 order of magnitude lower than those of JH II. JH I, II and III acids were also detected, but at levels generally lower than the corresponding JHs. JH titers in field collected reproductively inactive, gregarious adult monarchs, were 1–2 orders of magnitude lower than those of laboratory reared reproductively active monarchs. JH II was again the predominant JH in these animals.     

Diapause phenomena in non-diapausing last instar larvae,pupae, and pharate adults of the Colorado beetle

From the first day of the last (fourth) larval instar no trace of juvenile hormone (JH) can be detected in the haemolymph by Galleria bioassay. Three specific diapause proteins, which are also found in diapausing adults, appear in the haemolymph. These proteins disappear towards the end of the pupal stage. Study of the ultrastructure of the fat body revealed the formation from lysosomes of proteinaceous bodies which are also characteristic for adult diapause. The behaviour of last instar larvae and pupae resembles that of prediapausing and diapausing adults respectively. Injection of synthetic JH delays the appearance of the diapause proteins in the haemolymph and of proteinaceous bodies in the fat body for 2 to 3 days. The absence of JH seems to trigger off these diapause phenomena.     

Juvenile hormone induced ovarian uptake of a female-specific blood protein in Oncopeltus fasciatus

An adult female-specific blood protein was demonstrated in Oncopeltus by gel electrophoresis. This protein is the major band in soluble yolk fractions. It is also present at substantial concentrations in the haemolymph of starved and diapausing adult females. Thus, the failure of the ovaries to form yolk under these conditions is characterized by an inability to remove vitellogenin from the blood. Application of a juvenile hormone analog (JHA) restored protein yolk deposition in starved and diapausing adult females. Whereas other blood proteins decreased no more than two-fold upon JHA treatment, the vitellogenin concentration decreased 20-fold in starved females. The vitellogenin concentration in the blood of diapausing females was not significantly affected by JHA, apparently because synthesis kept pace with ovarian uptake in this case.     

Independently regulated juvenile hormone activity and vitellogenesis in mosquitoes

Temporally distinct, head-mediated processes regulate vitellogenic development as well as juvenile hormone (JH)-mediated development of ovarian follicles of Aedes aegypti. In blood-fed adult mosquitoes, vitellogenic development is stimulated during the first day after blood is imbibed and JH secretion is stimulated 2 days later. JH secretion in recently ecdysed adult mosquitoes is stimulated during or shortly before ecdysis. These observations suggest that vitellogenesis follows blood-ingestion, whereas JH activity may secondarily be promoted by vitellogenesis. It may be that vitellogenesis and JH activity are mediated by different brain hormones     

Endocrine biology of the Painted Lady butterfly Vanessa cardui

Experiments dealing with the role of juvenile hormone, adipokinetic hormone and diuretic hormone in the Painted Lady butterfly Vanessa cardui are reported. The results demonstrate an important role of JH in the regulation of ovarian and colleterial gland development in females, and in the regulation of accessory gland, tubular gland and ejaculatory duct development in males. In addition, the presence of an adult reproductive diapause, characterized by decreased effective juvenile hormone haemolymph titres, is suggested. Evidence for the existence of both adipokinetic and diuretic hormones in the Painted Lady is also presented, as is data indicating that both hormones may be similar or identical to those previously described in Monarch butterflies. Owing to the above results, and the existence of an artificial diet suitable for mass rearing in the laboratory, the Painted Lady appears to be an excellent species for studies on adult lepidopteran neuroendocrinology.     

Juvenile hormone titer and morph-specific reproduction in the wing-polymorphic cricket,Gryllus firmus

Juvenile hormone titers and reproductive characteristics were measured in adult wing and flight-muscle morphs of the wing-polymorphic cricket, Gryllus firmus, during the first week of adulthood. This species has three morphs: one flight capable morph with fully-developed wings and fully-developed flight muscles [LW(F)], one flightless morph with fully-developed wings and histolyzed (non-functional) flight muscles [LW(H)], and another flightless morph with underdeveloped (short) wings and underdeveloped flight muscles (SW). Both flightless morphs [LW(H) and SW] had larger ovaries which contained a greater number of postvitellogenic eggs compared with the flight capable [LW(F)] morph. The juvenile hormone titer was significantly higher in SW compared with LW(F) females on days 3-7 of adulthood. On these days, the JH titer also was significantly higher in the other flightless morph, LW(H), compared with flight-capable [LW(F)] females as determined by one statistical test, but did not differ significantly by another test. The JH titer was positively correlated with ovarian mass or terminal oocyte length, but not with the number of post-vitellogenic eggs. This study is the first direct comparison of juvenile hormone titers in adult wing morphs of a wing-polymorphic insect. Results indicate that an elevated juvenile hormone titer may be at least partly responsible for one of the most distinctive features of wing-polymorphic species, the increased early fecundity of flightless females.     

Role of juvenile hormone metabolism during embryogenesis of the house cricket,Acheta domesticus

The duration of embryogenesis was 9.5 days for house crickets, Acheta domesticus (L.), reared at 35°C. The major route of juvenile hormone (JH) metabolism was ester hydrolysis. The level of α-naphthyl acetate (α-NA) esterase activity per mg wet weight remained relatively constant throughout embryogenesis and was similar to that of eggs dissected from the oviducts. JH esterase activity per mg wet weight was highest in the dissected and day-1 eggs, declined to one-third of this peak activity by day 5, and then remained unchanged through hatching. Two populations of esterases (130,000 and > 200,000 in molecular weight) which metabolized JH and α-NA were resolved in day-1 eggs by gel filtration chromatography. Specific JH esterase appeared by day 4 with a molecular weight of 200,000. Correlative evidence is presented from other insect species that supports a functional role for JH metabolism during embryo development.     

Synchrony of juvenile hormone-sensitive periods for internal and external development in last-instar larvae of Oncopeltus fasciatus

Breakdown of the moulting glands in Oncopeltus can be completely inhibited by topical application of a juvenile hormone analogue prior to day 2 of the fifth instar, and partially inhibited by application prior to day 4. The analogue-sensitive period for the inhibition of external metamorphosis is very similar to that for the inhibition of cell death in the moulting glands. A decline in response to the analogue between days 2 and 4 is correlated with rising ecdysteroid levels in the haemolymph. That this rise in ecdysteroids may be responsible for termination of the juvenile hormone-sensitive period is suggested by premature loss of sensitivity to the analogue by the moulting glands following premature exposure to 20-hydroxyecdysone.     

Identification of the juvenile hormones from adult Attacus atlas

Both juvenile hormone (JH) II and JH I have been identified in whole body extracts of the atlas moth, Attacus atlas. In 0–24 hr old adult males, levels of JH II were ∼ 15 ng/g while JH I titers were much lower (∼0.4 ng/g). In keeping with titer data obtained from other Saturniid moths, male animals contained substantially more JH than adult females, which contained <0.1 ng/g of predominantly JH II. Our results are in contrast to a prior study which did not detect any of the known JHs and hinted at the possible existence of a new JH active principle. The levels of JH I plus II we detected in A. atlas can easily account for the amount of biological activity observed in the prior study. Hence, we are confident that JH II and JH I constitute the only JHs of adult A. atlas.     

Simultaneous determination of molting and juvenile hormone titers of the greater wax moth

A simple and rapid extraction procedure was developed to determine simultaneously the molting hormone (MH) and juvenile hormone (JH) activity in a single insect tissue sample. From the onset of the last larval stage to adult eclosion of the greater wax moth, Galleria mellonella, three JH peaks were noted: at the time of the sixth larval ecdysis, 1 day before the seventh larval ecdysis, and at the time of adult eclosion. Three MH peaks were recorded for the male: at 1 day before the sixth larval ecdysis, 1 day before the seventh larval ecdysis, and 2 days after pupation. In the female, a fourth peak was shown at the time of adult eclosion. This fourth peak exhibits the highest molting hormone activity of all samples, 1600 Musca units/g of fresh tissue or an equivalent of 5.6 μg/g of ecdysterone. Eighty per cent of this MH accumulated in the ovary. The significance of MH and JH titers as related to the endocrine regulation of development is discussed in the light of this finding.     

The juvenile hormone titer of Trichoplusia ni and its potential role in embryogenesis of the polyembryonic wasp CopidosomaFloridanum

The hemolymph juvenile hormone (JH) titer of third through fifth stadia Trichoplusia ni parasitized by the polyembryonic parasitoid, Copidosoma floridanum, was measured by radioimmunoassay and compared to the titers of unparasitized larvae. The JH titer of parasitized larvae fluctuated from 28 pg/μl to undetectable levels. Maximum levels of hormone were present at ecdysis to the fourth and fifth stadium, and at the prepupal stage. Qualitatively, similar fluctuations were observed in unparasitized larvae. However, the titers in unparasitized larvae were much lower than those of parasitized larvae in the third and early fourth stadia, and the titer fell to undetectable levels in the fifth stadium 24 h earlier (48 h) than in parasitized larvae (72 h). Preventing the JH titer from falling during the fourth and fifth stadia by topical application of (RS)-methoprene or JH II had a juvenilizing effect on parasitized T. ni, and inhibited C. floridanum embryo morphogenesis. The effect of exogenous methoprene and JH on C. floridanum development depended on timing of application and dosage. Application of 100 pmol per day of methoprene beginning at 2 h of the host fourth stadium, prior to the large drop in the endogenous JH titer, inhibited morphogenesis in the majority of C. floridanum embryos. Application of methoprene at later times of host development did not inhibit morphogenesis although other developmental alterations were observed. The potential significance of host JH and ecdysteroid titers on polyembryonic development are discussed.     

Haemolymph juvenile hormone III esterase activity during adult life of female and male crickets,Gryllus bimaculatus de Geer (Ensifera,Gryllidae)

Using an in vitro method, juvenile hormone III degradation was studied in the plasma of adult female and male crickets, Gryllus bimaculatus. The primary route of juvenile hormone III metabolism in cricket haemolymph is ester hydrolysis to juvenile hormone III acid by juvenile hormone III esterase. Juvenile hormone III esterase activity in females' haemolymph is low just after imaginal moulting. A sharp peak of enzyme activity is observed on day 3 after emergence, and two subsequent peak values occur in older animals. Plasma juvenile hormone III esterase activity in freshly ecdysed males is also low, but increases rapidly thereafter. Another increase in enzyme activity is observed in older males. The fluctuations in juvenile hormone III esterase activity are discussed in correlation with changes in haemolymph volume, haemolymph protein content, haemolymph juvenile hormone III titer, and the rates of juvenile hormone III biosynthesis in vitro of the corpora allata.     

Endocrine influence upon the development of vitellogenic competency in Oncopeltus fasciatus

Immunoelectrophoretic analysis of the blood of precocious adult females of Oncopeltus demonstrated the presence of the A form of vitellogenin but no detectable AB form, the form present in mature adult-female haemolymph. Although the appearance of the AB form could be induced by topical treatment of precocious adult females with juvenile hormone, no yolk deposition was induced in these females. Histological examination revealed that the ovaries of precocious adult females reached only a previtellogenic stage of development with or without treatment with juvenile hormone. Topical treatment of normal larvae and adults with the hormone demonstrated that vitellogenin synthesis could not be induced with juvenile hormone treatment alone until after adult emergence. Since the precocious adult females are chronologically younger than normal last-stage larval instars, we suggest that a transition period during which juvenile hormone is absent (i.e. the precocious moult in treated animals; the final moult in control animals) must occur before some tissue of the precocious or normal adult females, presumably the fat body, becomes competent to respond to further exposure to the hormone by making the AB form of vitellogenin. The ovary requires a similar transition period prior to becoming capable of depositing vitellogenin; however, the times when the fat body and the ovary become competent to respond to the transition period differ.     

Suppression of allatotropin simulates reproductive diapause in the mosquito Culex pipiens

The cessation of juvenile hormone (JH) production is a key endocrine event that halts ovarian development and hence initiates diapause in females of the mosquito, Culex pipiens. The shutdown in endocrine activity of the corpora allata (CA), the source of JH, was manifested in the smaller size of CA in females reared under short daylengths (diapause) compared to those reared under long daylengths (nondiapause), as well as in low expression of the mRNA encoding allatotropin, the neuropeptide that promotes JH biosynthesis in the CA. Genes encoding both allatotropin and allatostatin were identified in C. pipiens, but only expression levels of allatotropin differed in the two types of females. Knockdown of allatotropin mRNA using RNA interference in females programmed for nondiapause resulted in a cessation of ovarian development akin to diapause. This arrest in development could be reversed with an application of JH. Our results thus suggest that suppression of allatotropin is a critical link in regulating the shutdown of the CA during diapause.     

Juvenile hormone induces ovarian development in diapausing cave-dwelling Drosophila species

Drosophila grisea and macroptera were collected in caves overwintering as adults. The females remained in a state of reproductive diapause which extended until May for macroptera and until July for grisea, whereas the males of both species had mature sperm at all times. Termination of the reproductive diapause under laboratory conditions was accomplished in grisea by exposing them to 14 hr of illumination daily and in macroptera by increasing the temperature to 20°C. Topical application of juvenile hormone (JH) on diapausing grisea caused a prompt termination of diapause and maturation of oöcytes within 10 days. Yolk proteins were found in the haemolymph of diapausing flies but not in their ovaries. In the JH-treated flies, yolk proteins were found in both the haemolymph and the ovaries, suggesting that in this species JH regulates the uptake of yolk proteins.     

Control of larval-pupal-adult molt in the moth Sesamia nonagrioides by juvenile hormone and ecdysteroids

Sesamia nonagrioides (Lepidoptera: Noctuidae) larvae reared under long day (LD; 16L:8D) conditions pupate after 5 or 6 larval instars, whereas under short day (SD; 12L:12D) conditions they undergo up to 12 additional molts before pupating. This extended period of repeated molting is maintained by high levels of juvenile hormone (JH). Previous work demonstrated that both LD and SD larvae decapitated in the 6th instar pupate but further development is halted. By contrast, about one-third of SD larvae from which only the brain has been removed, undergo first a larval molt, then pupate and subsequently developed to the adult stage. Debrained LD larvae molt to larvae exceptionally but regularly pupate and produce adults. Implanted brains may induce several larval molts in debrained recipient larvae irrespectively of the photoperiodic conditions. The results of present work demonstrate that the prothoracic glands (PGs) and the corpora allata (CA) of debrained larvae continue to produce ecdysteroids and JHs, respectively. PGs are active also in the decapitated larvae that lack JH, consistent with the paradigm that CA, which are absent in the decapitated larvae, are the only source of this hormone. Completion of the pupal-adult transformation in both LD and SD debrained insects demonstrates that brain is not crucial for the development of S. nonagrioides but is required for diapause maintenance. Application of JH to headless pupae induces molting, presumably by activating their PGs. It is likely that JH plays this role also in the induction of pupal-adult transformation in debrained insects. Application of the ecdysteroid agonist RH 2485 (methoxyfenozide) to headless pupae also elicits molting: newly secreted cuticle is in some cases thin and indifferent, in other cases it bears distinct pupal or adult features.