Hemolymph Juvenile Hormone Titers Research Articles

Flight and flight energy accumulation related to the daily rhythm of juvenile hormone titer in the wing‐dimorphic cricket Velarifictorus aspersus

AbstractThe cricket Velarifictorus aspersus (Walker) (Orthoptera: Gryllidae) exhibits a wing dimorphism mediating two distinct life‐history strategies during early adulthood; long‐winged (LW) females mainly invest resources in flight, whereas short‐winged (SW) females invest more heavily in reproduction than LW females. To understand the relationships among juvenile hormone (JH), flight capacity, and energy accumulation, the daily rhythm of JH titer and flight activity, the concentrations of flight energy substances, and the influence of exogenous JH on flight activity and flight energy substances were investigated in V. aspersus. The results showed that the hemolymph JH titer exhibited a high‐amplitude daily rhythm in 5‐day‐old LW V. aspersus females, and the JH titer rose about 10× at 18:00 hours (6 h before lights off) and dropped to baseline levels after lights off. The females flew inactively (i.e., they floated in the air) from 14:00 to 19:00 hours during the light period, whereas they had a stronger flight capacity in the initial photophase and before lights out. The flight ability of LW females with 20 μg μL−1 JH treatments was significantly improved during periods of flight inactivity. After the peak JH titer, the concentrations of glycogen and trehalose in the flight muscles increased significantly. The LW females treated with JH during the inactive flight period had a significant increase in glycogen after 1 h and trehalose after 3 h. These results suggested that the high‐amplitude daily rhythm of the JH titer was correlated with accumulation of flight energy substance and flight capacity; that is, an elevated JH titer promoted the biosynthesis of flight energy substances, resulting in improvement of flight capacity.

Juvenile hormone interacts with multiple factors to modulate aggression and dominance in groups of orphan bumble bee (Bombus terrestris) workers

Juvenile hormone (JH) is a key regulator of insect development and reproduction. Given that JH commonly affects adult insect fertility, it has been hypothesized to also regulate behaviors such as dominance and aggression that are associated with reproduction. We tested this hypothesis in the bumble bee Bombus terrestris for which JH has been shown to be the major gonadotropin. We used the allatoxin Precocene-I (P-I) to reduce hemolymph JH titers and replacement therapy with the natural JH to revert this effect. In small orphan groups of workers with similar body size but mixed treatment, P-I treated bees showed lower aggressiveness, oogenesis, and dominance rank compared with control and replacement therapy treated bees. In similar groups in which all bees were treated similarly, there was a clear dominance hierarchy, even in P-I and replacement therapy treatment groups in which the bees showed similar levels of ovarian activation. In a similar experiment in which bees differed in body size, larger bees were more likely to be dominant despite their similar JH treatment and ovarian state. In the last experiment, we show that JH manipulation does not affect dominance rank in groups that had already established a stable dominance hierarchy. These findings solve previous ambiguities concerning whether or not JH affects dominance in bumble bees. JH positively affects dominance, but bees with similar levels of JH can nevertheless establish dominance hierarchies. Thus, multiple factors including JH, body size, and previous experience affect dominance and aggression in social bumble bees.

Methyl farnesoate epoxidase (mfe) gene expression and juvenile hormone titers in the life cycle of a highly eusocial stingless bee, Melipona scutellaris

In social insects, juvenile hormone (JH) has acquired novel functions related to caste determination and division of labor among workers, and this is best evidenced in the honey bee. In contrast to honey bees, stingless bees are a much more diverse group of highly eusocial bees, and the genus Melipona has long called special attention due to a proposed genetic mechanism of caste determination. Here, we examined methyl farnesoate epoxidase (mfe) gene expression, encoding an enzyme relevant for the final step in JH biosynthesis, and measured the hemolymph JH titers for all life cycle stages of Melipona scutellaris queens and workers. We confirmed that mfe is exclusively expressed in the corpora allata. The JH titer is high in the second larval instar, drops in the third, and rises again as the larvae enter metamorphosis. During the pupal stage, mfe expression is initialy elevated, but then gradually drops to low levels before adult emergence. No variation was, however, seen in the JH titer. In adult virgin queens, mfe expression and the JH titer are significantly elevated, possibly associated with their reproductive potential. For workers we found that JH titers are lower in foragers than in nurse bees, while mfe expression did not differ. Stingless bees are, thus, distinct from honey bee workers, suggesting that they have maintained the ancestral gonadotropic function for JH. Hence, the physiological circuitries underlying a highly eusocial life style may be variable, even within a monophyletic clade such as the corbiculate bees.

Analysis of the Classical Model of Juvenile Hormone Control of Wing Polymorphism in the CricketVelarifictorus aspersus(Orthoptera: Gryllidae)

The classical model of juvenile hormone (JH)-mediated wing polymorphism was proposed in the early 1960s, and numerous studies have tested this model using exogenous hormone treatment. However, this indirect method may yield misleading results, necessitating direct quantification of JH titers in insects. In this study, the effects of JH application on wing development and comparisons of hemolymph JH titers between presumptive long-winged (LW) and short-winged (SW) nymphs in the cricket Velarifictorus aspersus (Walker) are investigated. The results show that treatment of presumptive LW nymphs with high-dose JH III significantly decreases the percentage of macropterous individuals during the penultimate and first half of the last stadia, indicating that exogenous JH III can suppress wing development in V. aspersus and that the last two stadia are critical periods for wing determination. However, application of precocene to presumptive SW nymphs does not induce macropterizing effects, and JH III titers are not significantly higher in presumptive SW females than in presumptive LW females during the critical period of wing development. Therefore, this model may not be applicable to this species and the brachypterizing effects associated with JH application may be induced by affecting other innate factors rather than by direct JH titers in hemolymph.

The role of juvenile hormone in dominance behavior, reproduction and cuticular pheromone signaling in the caste-flexible epiponine wasp, Synoeca surinama.

BackgroundThe popular view on insect sociality is that of a harmonious division of labor among two morphologically distinct and functionally non-overlapping castes. But this is a highly derived state and not a prerequisite for a functional society. Rather, caste-flexibility is a central feature in many eusocial wasps, where adult females have the potential to become queens or workers, depending on the social environment. In non-swarming paper wasps (e.g., Polistes), prospective queens fight one another to assert their dominance, with losers becoming workers if they remain on the nest. This aggression is fueled by juvenile hormone (JH) and ecdysteroids, major factors involved in caste differentiation in most eusocial insects. We tested whether these hormones have conserved aggression-promoting functions in Synoeca surinama, a caste-flexible swarm-founding wasp (Epiponini) where reproductive competition is high and aggressive displays are common.ResultsWe observed the behavioral interactions of S. surinama females in field nests before and after we had removed the egg-laying queen(s). We measured the ovarian reproductive status, hemolymph JH and ecdysteroid titers, ovarian ecdysteroid content, and analyzed the cuticular hydrocarbon (CHC) composition of females engaged in competitive interactions in both queenright and queenless contexts. These data, in combination with hormone manipulation experiments, revealed that neither JH nor ecdysteroids are necessary for the expression of dominance behaviors in S. surinama. Instead, we show that JH likely functions as a gonadotropin and directly modifies the cuticular hydrocarbon blend of young workers to match that of a reproductive. Hemolymph ecdysteroids, in contrast, are not different between queens and workers despite great differences in ovarian ecdysteroid content.ConclusionsThe endocrine profile of S. surinama shows surprising differences from those of other caste-flexible wasps, although a rise in JH titers in replacement queens is a common theme. Extensive remodeling of hormone functions is also evident in the highly eusocial bees, which has been attributed to the evolution of morphologically defined castes. Our results show that hormones which regulate caste-plasticity can lose these roles even while caste-plasticity is preserved.Electronic supplementary materialThe online version of this article (doi:10.1186/s12983-014-0078-5) contains supplementary material, which is available to authorized users.

Juvenile Hormone Biosynthesis Gene Expression in the corpora allata of Honey Bee (Apis mellifera L.) Female Castes

Juvenile hormone (JH) controls key events in the honey bee life cycle, viz. caste development and age polyethism. We quantified transcript abundance of 24 genes involved in the JH biosynthetic pathway in the corpora allata-corpora cardiaca (CA-CC) complex. The expression of six of these genes showing relatively high transcript abundance was contrasted with CA size, hemolymph JH titer, as well as JH degradation rates and JH esterase (jhe) transcript levels. Gene expression did not match the contrasting JH titers in queen and worker fourth instar larvae, but jhe transcript abundance and JH degradation rates were significantly lower in queen larvae. Consequently, transcriptional control of JHE is of importance in regulating larval JH titers and caste development. In contrast, the same analyses applied to adult worker bees allowed us inferring that the high JH levels in foragers are due to increased JH synthesis. Upon RNAi-mediated silencing of the methyl farnesoate epoxidase gene (mfe) encoding the enzyme that catalyzes methyl farnesoate-to-JH conversion, the JH titer was decreased, thus corroborating that JH titer regulation in adult honey bees depends on this final JH biosynthesis step. The molecular pathway differences underlying JH titer regulation in larval caste development versus adult age polyethism lead us to propose that mfe and jhe genes be assayed when addressing questions on the role(s) of JH in social evolution.

Argonaute 1 is indispensable for juvenile hormone mediated oogenesis in the migratory locust, Locusta migratoria

Juvenile hormone (JH) is the primary hormone controlling vitellogenesis and oocyte maturation in the migratory locust Locusta migratoria, an evolutionarily primitive insect species with panoistic ovaries. However, molecular mechanisms of locust oogenesis remain unclear and the role of microRNA (miRNA) in JH mediated locust vitellogenesis and oocyte maturation has not been explored. Using miRNA sequencing and quantification with small RNA libraries derived from fat bodies of JH-deprived versus JH analog-exposed female adult locusts, we have identified 83 JH up-regulated and 60 JH down-regulated miRNAs. QRT-PCR validation has confirmed that transcription of selected miRNAs responded to JH administration and correlated with changes in endogenous hemolymph JH titers. Depletion of Argonaute 1 (Ago1), a key regulator of miRNA biogenesis and function by RNAi in female adult locusts dramatically decreased the expression of vitellogenin (Vg) and severely impaired follicular epithelium development, terminal oocyte maturation and ovarian growth. Our data indicate that Ago1 and Ago1-dependent miRNAs play a crucial role in locust vitellogenesis and egg production.

Molecular characterization and gene expression of juvenile hormone binding protein in the bamboo borer, Omphisa fuscidentalis

Juvenile hormone (JH) plays an important role in many physiological processes in insect development, diapause and reproduction. An appropriate JH titer in hemolymph is essential for normal development in insects. Information concerning its carrier partner protein, juvenile hormone binding protein (JHBP), provides an alternative approach to understanding how JH regulates metamorphosis. In this study, we cloned and sequenced the Omphisa juvenile hormone binding protein (OfJHBP). The full-length OfJHBP cDNA sequence is comprised of 849 nucleotides with an open reading frame of 726bp encoding 242 amino acids. The molecular mass of the protein was estimated to be 26.94kDa. The deduced protein sequence of OfJHBP showed moderate homology with the lepidopteran, Heliothis virescens JHBP (52% amino acid identity) and lower homology with the Bombyx mori JHBP (45%) and the Manduca sexta JHBP (44%). The OfJHBP was expressed mainly in the fat body. OfJHBP transcripts in the fat body was moderately high during 3rd, 4th and 5th instars, then rapidly increased, reaching a peak during early diapause. The expression remained high in mid-diapause, then decreased in late-diapause until the pupal stage. Both juvenile hormone analog (JHA), methoprene, 20-hydroxyecdysone (20E) exhibited a similar stimulatory pattern in OfJHBP expression of diapausing larvae. OfJHBP mRNA levels gradually increased and showed a peak of gene expression on the penultimate, then declined to low levels in the pupal stage. For in vitro gene expression, both of JHA and 20E induced OfJHBP mRNA expression in fat body. Fat body maintenance in vitro in the presence of 0.1μg/50μl JHA induced OfJHBP mRNA expression to high levels within the first 30min whereas 0.1μg/50μl 20E induced gene expression at 120min. To study the synergistic effect of these two hormones, fat body was incubated in vitro with 0.1μg/50μl JHA or 0.1μg/50μl 20E or a combination of both hormone for 30min. Induction of OfJHBP expression by JHA and 20E was significantly greater than that of either hormone alone. These results should contribute to our understanding of how JHBP and JH regulate the termination of larval diapause in the bamboo borer.

The influence of drone physical condition on the likelihood of receiving vibration signals from worker honey bees, Apis mellifera

Honey bee workers will perform vibration sig- nals on adult drones, which respond by increasing the time spent receiving trophallaxis. Because trophallaxis provides the proteins for sexual maturation, workers could direct vibration signals towards drones showing certain physical characteristics, potentially influencing drone development and colony reproductive output. We examined the influence of drone condition on the likelihood of receiving vibration signals by comparing body weight, protein concentrations, and hemolymph juvenile hormone (JH) titers between drones that received the vibration signal and same-age, non- vibrated controls. Vibrated and control drones did not differ in total body weight, abdomen weight, abdomen-to-body weight ratio, total protein concentrations, or hemolymph JH titers. In contrast, vibrated drones had significantly lower thorax weight and smaller thorax-to-body weight ratios compared with controls. Because relative thorax weight may affect flight ability and mating success, workers could use the vibration signal to increase the care received by less developed drones, potentially contributing to the production of greater numbers of competitive males. However, the differences in thorax weights, while significant, were very small, and it is unknown how such slight differences might be assessed by workers or affect drone performance. Nev- ertheless, vibration signals performed on drones may pro- vide opportunities for exploring the effect of the quality of reproductive individuals on caste interactions in honey bees.

Juvenile hormone titers and caste differentiation in the damp-wood termite Hodotermopsis sjostedti (Isoptera, Termopsidae)

Termites are social insects, presenting morphologically distinct castes, performing specific tasks in the colony. The developmental processes underlying caste differentiation are mainly controlled by juvenile hormone (JH). Although many fragmentary data support this fact, there was no comparative work on JH titers during the caste differentiation processes. In this study, JH titer variation was investigated using a liquid chromatography-mass spectrometry (LC-MS) quantification method in all castes of the Japanese damp-wood termite Hodotermopsis sjostedti, especially focusing on the soldier caste differentiation pathway, which was induced by treatment with a JH analog. Hemolymph JH titers fluctuated between 20 and 720 pg/μl. A peak of JH was observed during molting events for the pseudergate stationary molt and presoldier differentiation, but this peak was absent prior to the imaginal molt. Soldier caste differentiation was generally associated with high JH titers and nymph to alate differentiation with low JH titers. However, JH titer rose in females during alate maturation, probably in relation to vitellogenesis. In comparison, JH titer was surprisingly low in neotenics. On the basis of these results in both natural and artificial conditions, the current model for JH action on termite caste differentiation is discussed and re-appraised.

Hemolymph Juvenile Hormone Titers in Worker Honey Bees under Normal and Preswarming Conditions

Swarming is an important mechanism by which honey bee, Apis mellifera L., colonies reproduce, yet very little is known about the physiological changes in workers that are preparing to swarm. In this study, we determined the endocrine status of worker honey bees in preswarming colonies and in normal (nonswarming) colonies. Juvenile hormone (JH) titers in worker bees were similar in both groups before queen cells were present, but they became significantly lower in preswarming colonies compared with normal colonies when queen cells occurred in preswarming colonies. The lower JH titers in the preswarming colonies suggest that behavioral development is delayed in these colonies, consistent with previous reports that preswarming colonies have reduced foraging activities. Understanding the endocrine status of bees preparing for swarming will help us to better understand the biology of swarming.

Hemolymph Juvenile Hormone Titers in Worker Honey Bees under Normal and Preswarming Conditions

Swarming is an important mechanism by which honey bee, Apis mellifera L., colonies reproduce, yet very little is known about the physiological changes in workers that are preparing to swarm. In this study, we determined the endocrine status of worker honey bees in preswarming colonies and in normal (nonswarming) colonies. Juvenile hormone (JH) titers in worker bees were similar in both groups before queen cells were present, but they became significantly lower in preswarming colonies compared with normal colonies when queen cells occurred in preswarming colonies. The lower JH titers in the preswarming colonies suggest that behavioral development is delayed in these colonies, consistent with previous reports that preswarming colonies have reduced foraging activities. Understanding the endocrine status of bees preparing for swarming will help us to better understand the biology of swarming.

Parasitism-induced effects of Glyptapanteles liparidis (Hym., Braconidae) on the juvenile hormone titer of its host, Lymantria dispar: the role of the parasitoid larvae

Parasitization by the gregarious larval endoparasitoid Glyptapantles liparidis induces a dramatic increase in the hemolymph juvenile hormone (JH) titer (especially JH III) of its host larva, Lymantria dispar. Here, we investigated the role of the parasitoid larvae in JH synthesis and release by in vitro and in vivo experiments. GC-MS analyses confirmed that the rising hemolymph JH titer coincided with the time at which the parasitoids molt to the second larval instar. Peak values in host hemolymph titers were observed prior to parasitoid emergence, and titers dropped to negligible levels within 24 h after parasitoid emergence. Whole body extracts from excised second instar parasitoids yielded JH III and trace amounts of JH II. The in vitro secretory activity of the corpora allata (CA) of L. dispar larvae was not enhanced by parasitization. When the host's CA were separated by neck ligation, we found elevated JH III titers, but no JH II in the hemolymph of the posterior section, which contained the parasitoids. Parasitoids that were kept in in vitro culture produced and released only JH III. The parasitoids’ ability to secrete JH and to molt independently from their host's molting cycles indicates that at least second instar parasitoids are hormonally self-reliant.

A morph-specific daily cycle in the rate of JH biosynthesis underlies a morph-specific daily cycle in the hemolymph JH titer in a wing-polymorphic cricket

A previous study documented a high amplitude, morph-specific daily cycle in the hemolymph JH titer in the wing-polymorphic cricket, Gryllus firmus. The JH titer rose and fell 10–20 fold in the flight-capable [LW(f), long-winged] morph during the late-photophase-early scotophase, while it was relatively constant during that time in the flightless (SW, short-winged) morph. In the present study we documented a dramatic morph-specific daily cycle in the in vitro rate of juvenile hormone (JH) biosynthesis that was tightly correlated with the hemolymph JH titer on days 5–7 of adulthood. Biosynthetic rates rose and fell 1–2 fold between the late photophase-early scotophase on each of days 5–6 and 6–7 of adulthood in the LW(f) morph, while biosynthetic rates were relatively constant during this period in the flightless, short-winged morph (SW), except for a slight dip in the rate of biosynthesis late in the photophase on these days. Similar morph-specific patterns of JH biosynthesis were observed whether rates were measured on corpora allata attached to corpora cardiaca in males or females, or on corpora allata alone. Hemolymph juvenile hormone esterase activity was significantly higher in the LW(f) vs. the SW morph during the beginning of scotophase, when the JH titer is decreasing rapidly in the LW(f) morph. Results indicate that the morph-specific daily cycle in the JH titer in G. firmus is primarily regulated by a morph-specific daily cycle in the rate of JH biosynthesis and to a lesser degree by hemolymph JH esterase activity. This is the first documentation of a diurnal cycle in the rate of JH biosynthesis in any insect, or a daily cycle in the rate of JH biosynthesis that is correlated with a specific morph in a polymorphic species. Results have important implications for the endocrine regulation of dispersal polymorphism, circadian rhythms of insect hormone titers and their regulators, and general studies of the JH titer and its regulation in insects.

The hemolymph JH titer exhibits a large-amplitude, morph-dependent, diurnal cycle in the wing-polymorphic cricket, Gryllus firmus

The hemolymph juvenile hormone (JH) titer was measured in over 500 flight-capable and flightless, adult female Gryllus firmus at 3–6 h intervals during each of days 2–8 of adulthood. The flight-capable morph exhibited a large-amplitude daily cycle in the hemolymph JH titer, while the flightless morph exhibited a barely perceptible cycle. The JH titer cycle was observed on all days in the flight-capable morph, but the large amplitude cycle (>15–20 fold increase in mean titer; >100-fold increase in some individuals), began on day 5. For both the large and small amplitude cycles, the JH titer peaked near the end of the photophase–beginning of the scotophase. The hemolymph ecdysteroid titer did not exhibit a corresponding large amplitude daily cycle, although a low amplitude cycle (1–3-fold change) was seen in both morphs. The large magnitude rise in the JH titer in the flight-capable morph during the photophase was not due to decreased hemolymph volume or JH degradation. Daily cycles in the JH titer may be common, but may have gone unnoticed in other insect species due to restricted temporal sampling. Failure to identify these cycles can result in substantial errors in inferring biological roles for JH. Because JH regulates flight behaviors, morph-specific daily cycles in the JH titer may be especially common in dispersal-polymorphic insects, in which flight is restricted to one morph during a limited period of the day or night. However, because JH regulates numerous biological traits, analogous cycles may be common in insects exhibiting other types of complex (e.g. caste or phase) polymorphism, in which morphs differ in a biological characteristic that is restricted to a specific period of the photophase or scotophase.

In vivo effects of allatostatins in crickets, Gryllus bimaculatus (Ensifera: Gryllidae)

Two types of cricket allatostatins, Grb-AST A1 and Grb-AST B1, were injected into adult female crickets three times each day on days 0–3 and once on day 4 after adult emergence to test their activity in vivo. On day 4, body weight, ovary weight, number of eggs per ovary, length of the terminal oocytes, ovarian ecdysteroid biosynthesis, and hemolymph titers of ecdysteroids were lower in the allatostatin-injected animals compared with untreated and Ringer-injected controls. Effects of the injected allatostatins on hemolymph juvenile hormone titers were inhomogeneous, and no differences were found in the capacity of the corpora allata to produce juvenile hormone ex vivo. The hemolymph titers of yolk proteins (vitellogenins) were almost twice as high in the allatostatin-injected animals as in the control animals. The effects of the injected allatostatins and their interactions with the endocrine system of the animal are discussed. Arch. Insect Biochem. Physiol. 38:32– 43, 1998. © 1998 Wiley-Liss, Inc.

In vivo effects of allatostatins in crickets,Gryllus bimaculatus (Ensifera: Gryllidae)

Two types of cricket allatostatins, Grb-AST A1 and Grb-AST B1, were injected into adult female crickets three times each day on days 0–3 and once on day 4 after adult emergence to test their activity in vivo. On day 4, body weight, ovary weight, number of eggs per ovary, length of the terminal oocytes, ovarian ecdysteroid biosynthesis, and hemolymph titers of ecdysteroids were lower in the allatostatin-injected animals compared with untreated and Ringer-injected controls. Effects of the injected allatostatins on hemolymph juvenile hormone titers were inhomogeneous, and no differences were found in the capacity of the corpora allata to produce juvenile hormone ex vivo. The hemolymph titers of yolk proteins (vitellogenins) were almost twice as high in the allatostatin-injected animals as in the control animals. The effects of the injected allatostatins and their interactions with the endocrine system of the animal are discussed. Arch. Insect Biochem. Physiol. 38:32– 43, 1998. © 1998 Wiley-Liss, Inc.

Hemolymph juvenile hormone titers in pupal and adult stages of southwestern corn borer [ Diatraea grandiosella (pyralidae)] and relationship with egg development

Juvenile hormones I, II and III were monitored in hemolymph of pupal and adult stages of various ages of Diatraea grandiosella females. JH III was the predominant homologue followed by JH II, and JH I was rarely detectable. At day 5 after pupation, no JH was detectable. JH titers increased from 7.5 days after pupation to a peak of 24.8 ng ml −1 JH II and 26 ng ml −1 JH III at adult emergence and then declined to low levels by 24 h after emergence. Ovarian development in D. grandiosella parallels changes in hemolymph JH titers, but the role of JH in vitellogenesis is unclear since the time of vitellogenesis initiation has yet to be determined. No apparent vitellogenin deposition was observed in eggs 5 days after pupation. Some oocytes were partially vitellogenic by 7.5 days after pupation and oocytes continued to grow afterwards, but no oocytes were chorionated during the pupal stage. Chorionated oocytes were observed in 24-h-old female moths. Juvenile hormone is essential for chorion formation in this species, because decapitated pupae treated with 10 μg JH III in corn oil developed chorionated oocytes while decapitated pupae treated with corn oil did not.

Vitellogenin and egg production in the moth,Heliothis virescens

The characteristics of vitellogenin (Vg) and the relationship between Vg production and egg production in the tobacco budworm, Heliothis virescens, were studied. The relationship between Vg production and juvenile hormone (JH) and the impact of mating on Vg and egg production were also investigated. Vg appears in the hemolymph of H. virescens about 6 h after moth eclosion. Vg may be separated into two apoproteins (ApoVg-I and ApoVg-II) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights were calculated to be 156,065 ± 800 for ApoVg-I and 39,887 ± 323 for ApoVg-II. SDS-PAGE analysis revealed that the female hemolymph Vg polypeptides appear to be identical to those from eggs but are absent in male hemolymph. Vg concentration was significantly higher in mated females than in virgin females of the same age at 48 h after emergence. Rates of egg production increased as Vg production increased; rates of egg production in mated females were significantly higher than those of virgin females at 48, 72, 96, and 120 h postemergence. Vg production is dependent on JH, because hemolymph from decapitated females lacked Vg while that of decapitated females treated with synthetic JH had Vg at levels comparable to similarly aged, normal H. virescens females. Hemolymph JH titers in mated females were significantly higher compared with those in virgin females at all sampling periods. The high JH level in mated females may explain the high Vg and egg production in mated H. virescens. Arch. Insect Biochem. Physiol. 34:287–300, 1997. © 1997 Wiley-Liss, Inc.

Role of the corpora allata and juvenile hormone in the control of imaginal diapause and reproduction in three species of locusts

To understand the hormonal regulation of imaginal diapause and ovarian development in three species of locusts, Nomadacris succincta, Nomadacris japonica and Locusta migratoria, juvenile hormone (JH) biosynthesis by the corpora allata (CA) in vitro and JH esterase activity in the hemolymph were measured. The two species of Nomadacris exhibited a positive correlation between oocyte growth and JH biosynthesis by CA in vitro, although the correlation was significant only in N. succincta. In contrast, L. migratoria showed a negative correlation, indicating a higher rate of JH synthesis associated with more suppressed ovarian development. In this species, however, the hemolymph JH titer was higher in reproductively active females than in sexually immature females, indicating a positive correlation between the hemolymph JH titer and ovarian development. The levels and temporal pattern of variation in mean hemolymph JH esterase activity were rather similar between reproductively active and inactive females in L. migratoria. It is thus unlikely that the hemolymph JH esterase plays an important role in the regulation of the hemolymph JH titers in this species. Topical application of 100 μg of methoprene stimulated ovarian development significantly in L. migratoria, whereas 450 μg failed to induce ovarian development or vitellogenesis in sexually immature females of N. succincta and N. japonica. Besides JH, there appear to be other factors present that are involved in the control of the termination of diapause in the two Nomadacris species.