Juvenile hormone catabolism and oviposition in the codling moth, Cydia pomonella, as functions of age, mating status, and hormone treatment.

In vitro catabolism of juvenile hormone (JH) in haemolymph of adult female Cydia pomonella was ascribed mainly to juvenile hormone esterase (JHE) activity. No significant differences were noted between virgin and mated females 0-96 h post-emergence. Changes in JHE activity did not appear dependent upon fluctuations in JH titre; conversely, changes in JHE activity could not explain the changes in JH titres. Maximal JHE activity was recorded at 24 h (331.47 +/- 47.25 pmol/h/microl; 355.93 +/- 36.68 pmol/h/microl, virgin; mated insects, respectively) and preceded the peak in JH titres at 48 h. Topical application of JH II (10 ng-10 microg) or fenoxycarb (50 ng) enhanced JHE activity up to 640 and 56%, respectively. Treatment upon emergence with 10 microg JH II induced enzymic activity for less than 24 h, and when 10 microg JH II or 50 ng fenoxycarb were applied, circulating JH titres returned to control levels within 24 h. Oviposition was highly sensitive to exogenous JH and declined significantly with dosages >100 pg. To allow a degree of oocyte maturation before JH treatment, the hormone was administered at 6, 12, 24, or 48 h post-emergence and/or females were mated. Neither measure "protected" the system; oviposition declined immediately after JH application.     

Regulation of juvenile hormone biosynthesis in Heliothis virescens by Manduca sexta allatotropin

In Heliothis virescens, reproduction is strictly dependent on juvenile hormone (JH). In females, mating induces a sharp increase in JH titers, which stimulates increased vitellogenin biosynthesis and higher rates of egg production. JH biosynthesis is presumably stimulated by production and/or release of stimulatory neuropeptides such as allatotropins. There is evidence that allatotropin of H. virescens may be structurally related to Manduca sexta allatotropin (Manse-AT). In a radiochemical in vitro assay, synthetic Manse-AT stimulated JH biosynthesis by corpora allata (CA) of virgin H. virescens females in a dose-dependent manner, but had no effect on CA activity in H. virescens males. In females, the CA showed a transient increase in sensitivity to Manse-AT shortly after mating. Several structurally related peptides stimulated CA activity to a similar extent as Manse-AT. Corpora allata activity was stimulated by a Ca2+ ionophore, A23187. A membrane-permeable Ca2+ chelator, BAPTA/AM, antagonized the stimulatory effects of Manse-AT, suggesting that Manse-AT may enhance CA activity by increasing intracellular Ca2+ concentration.     

Manduca sexta allatotropin and the in vitro biosynthesis of juvenile hormone by moth corpora allata: a comparison of Pseudaletia unipuncta females from two natural populations and two selected lines

We compared the effects of Manduca sexta allatotropin (Manse-AT) on the rate of in vitro juvenile hormone (JH) biosynthesis by the corpora allata (CA) of different-aged virgin females from migrant (Quebec) and non-migrant (Azores) populations of the armyworm, Pseudaletia unipuncta, as well as from early- and late-calling lines selected from the Quebec population. There was a significant age x strain interaction, with the observed rates of JH biosynthesis in early adult life closely reflecting strain-specific differences in the age at onset of calling. In considering data for all ages combined, treatment of CA with Manse-AT resulted in a significant increase in the rate of JH biosynthesis for all but the Late strain, although significant differences for this strain were detected at certain ages. The CA of females from the Azores strain showed the strongest stimulation, with those of 0- and 1-day-old individuals displaying a singularly high degree of sensitivity. Selection for early- and late-calling lines resulted in significant differences in the temporal patterns of JH biosynthesis but did not markedly affect the sensitivity of the CA to Manse-AT. These findings are discussed within the context of the age-related differences observed in the rates of in vitro JH biosynthesis and JH haemolymph titers previously reported in comparisons of the Quebec and Azorean strains of the true armyworm.     

Regulation of JH epoxide hydrolase versus JH esterase activity in the cabbage looper, Trichoplusia ni, by juvenile hormone and xenobiotics

JH III esterase and JH III epoxide hydrolase (EH) in vitro activity was compared in whole body Trichoplusia ni homogenates at each stage of development (egg, larva, pupa and adult). While activity of both enzymes was detected at all ages tested, JH esterase was significantly higher than EH activity except for day three of the fifth (last) stadium (L5D3). For both enzymes, activity was highest in eggs. Adult virgin females had 4.6- and 4.0-fold higher JH esterase and EH activities, respectively, than adult virgin males. JH III metabolic activity also was measured in whole body homogenates of fifth stadium T. ni that were fed a nutritive diet (control) or starved on a non-nutritive diet of alphacel, agar and water. With larvae that were starved for 6, 28 and 52 h, EH activity per insect equivalent was 48%, 5% and 1%, respectively, of the control insects. At the same time points, JH esterase activity levels in starved T. ni were 29%, 4% and 3% of that of insects fed the nutritive diet. Selected insect hormones and xenobiotics were administered topically or orally to fifth stadium larvae for up to 52 h, and the effects on whole body EH and JH esterase activity analyzed. JH III increased the JH III esterase activity as high as 2.2-fold, but not the JH III EH activity. The JH analog, methoprene, increased both JH esterase and EH activity as high as 2.5-fold. The JH esterase inhibitor, 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP), had no impact on EH activity. The epoxides trans- and cis-stilbene oxide (TSO and CSO) in separate experiments increased the EH activity approximately 2.0-fold. TSO did not alter JH esterase levels when topically applied, but oral administration reduced activity to 70% of the control at 28 h, and then increased the activity 1.8-fold at 52 h after the beginning of treatment. CSO had no effect on JH esterase activity. Phenobarbital increased EH activity by 1.9-fold, but did not change JH esterase levels. Clofibrate and cholesterol 5alpha,6alpha-epoxide had no effect on EH. JH esterase activity also was not affected by clofibrate, but cholesterol 5alpha,6alpha-epoxide reduced the JH esterase activity to 60-80% of the control. The biological significance of these results is discussed.     

The effect of enforced virginity and subsequent mating on the activity of the corpus allatum of Periplaneta americana measured in vitro, as related to changes in the rate of ovarian maturation

ABSTRACT. Female P. americana, reared with males from the time of adult emergence, mated on the 4th–5th day after metamorphosis, produced the first ootheca on the 8th or 9th day, and then produced successive oothecae at intervals of 3.0 days, whereas, only 50% of virgin females had produced their first ootheca by the 28th day after adult emergence. Examination of the ovaries indicated that oocyte development is normal in virgins until shortly after the time when they first become receptive to males. When mating was not allowed there was a dramatic reduction in the rate of vitellogenic growth of the terminal batch of oocytes which persisted until mating was allowed, and was often accompanied by resorption of a percentage of the oocytes. Short-term, in vitro, radiochemical assay of juvenile hormone (JH III) biosynthesis by corpora allata (CA) showed that, in females reared with males, the cycles of ovarian development are accompanied by regular pulses of CA activity. There is a small, possibly preparatory peak of JH III biosynthesis before vitellogenesis of the first wave of oocytes, followed by a larger peak of JH III production during vitellogenesis of this batch of eggs and one peak of CA activity between ovulation of each subsequent wave of oocytes. Activities as low as 0.25 pmol C₁₆JH/CA pair/h and as high as 48.38 pmol/CA pair/h were observed in CA from mated females after the onset of cyclic activity. Stimuli received during mating are somehow responsible for the cyclic activity of the CA, for when females were subjected to enforced virginity the first small peak was normal but the second peak was not fully realized and there was then a gradual decline in CA activity until approximately 2 weeks post-emergence. Thereafter the glands exhibited a more or less constant rate of JH biosynthesis (mean = 3.45 ± 0.32 pmol/CA pair/h.) When females were mated after 21 days of enforced virginity the activity of the CA was enhanced. By 48 h after mating the mean glandular activity was at least four times that found in virgins of the same age, and by 72 h rates as high as 40 pmol/CA pair/h were observed. This was followed by normal cyclic activity of the CA. The increase in rate of JH biosynthesis appears to result in a recommencement of oocyte development in these ‘delayed-mated’ females.     

Juvenile hormone III produced in male accessory glands of the longhorned beetle,Apriona germari,is transferred to female ovaries during copulation

We report on juvenile hormone (JH) biosynthesis in vitro by male accessory glands (MAGs) in the longhorned beetle, Aprionona germari, accompanied by the transfer of JH from males to females during copulation. JH was extracted from the MAGs and separated by reversed‐phase high‐performance liquid chromatography. JH III was identified as the major JH by gas chromatography–mass spectrometry. A radiochemical assay and a non‐radioactive method were used to measure the in vitro rate of JH biosynthesis by the MAGs. After 4 h of incubation with ³H‐methionine in the medium, the radioactivity in the MAGs substantially increased. In a separate assay, incubation of the MAGs with non‐radioactive methionine for 4 h resulted in a 39% increase in JH III. Seven‐day‐old males were injected with medium 199 containing ³H–methionine and 24 h later they were mated with virgin females. Hemolymph and the MAGs were collected from the mated males and hemolymph, ovaries and eggs were collected from the mated females for assaying radioactive JH. The radioactivity incorporated into JH in the MAGs was transferred to the females during copulation and later transferred into their eggs. Assayed 1 h after copulation, JH III level in the MAGs decreased 42% and the content of JH III in the male hemolymph did not change, whereas the content of JH III in the female hemolymph and ovaries both increased. © 2010 Wiley Periodicals, Inc.     

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

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

Role of juvenile hormone esterase and epoxide hydrolase in reproduction of the cotton bollworm, Helicoverpa zea

The role of juvenile hormone (JH) esterase (JHE) and epoxide hydrolase (EH) in reproduction of the cotton bollworm, Helicoverpa zea, was investigated. Peak emergence of male and female bollworm adults occurred early in the scotophase. Female adults were added to males in a 1:2 ratio, respectively, at the beginning of the first photophase after emergence (d0). The highest oviposition rates for mated females were noted on d 2-4. The in vitro JH III esterase and JH III EH activity was measured in whole body homogenates of virgin and mated females from d0 to d8 post-emergence. Maximal JHE activity for virgin females occurred on d2 (1.09+/-0.14(+/-1 SEM) nmol of JH III degraded/min/mg protein), which was approximately twice that of mated females on the same day. The same results were observed for EH where the activity peaked on d2 at 0.053+/-0.003 as compared to 0.033+/-0.003 nmol of JH III degraded/min/mg protein, respectively. By d4, both JHE and JH EH activities declined significantly in virgin and mated females and were the same through d7. The developmental changes and effects of mating on JH degradation were similar when measured per insect. The highest levels of JHE and JH EH activity/min/mg protein in d2 virgin and mated females was found in ovaries followed by the carcass and then haemolymph; no EH activity was found in haemolymph as expected. For ovary, the JHE and JH EH activity was highest in virgin compared to mated females. The role of both enzymes in the regulation of reproduction is discussed.     

Influence of juvenile hormone and mating on oogenesis and oviposition in the codling moth,Cydia pomonella

Oogenesis in the codling moth, Cydia pomonella, and the role of juvenile hormones (JHs) were addressed. Rudimentary ovarian structures were recognisable in day 3–4 pupae, when haemolymph JH was still undetectable by coupled gas chromatography‐mass spectrometry in the selected ion mode (GC‐MS/SIM). The presence of developing oocytes was observed by light microscopy on day 8, coincident with very low JH titres (0.74 ± 0.05 ng/ml JH II). Chorionation was only evident upon emergence, following an increase in JH in the pharate adult (0h old: 4.71 ± 0.34 ng/ml JH II). Analysis of haemolymph from virgin and mated females indicated that JH II was predominant, with approximately equal and lower quantities of JHs I and III (3.3‐ to 5.0‐fold less). When pupae or newly emerged adults were treated with JH homologues, no alteration in ovarian protein content was apparent, but the JH mimetic, fenoxycarb, depressed the number of oocytes filling ≥ 50% follicular volume. Chorion deposition was stimulated by JHs I, II, or III (10 μg), but not by fenoxycarb (0.05 μg, 10 μg). Mating provided correct stimuli for enhanced choriogenesis and egg laying, and, since haemolymph JH titres were concomitantly elevated (approximately 2‐fold), it was postulated that the rise in JH elicited both these events. Application of JHs to virgin females, however, could not mimic mating; only increases in choriogenesis were induced: JH‐treatment of virgins (or mated insects) significantly decreased oviposition rates over 24 and 48 h and markedly reduced the life‐time total number of eggs. Arch. Insect Biochem. Physiol. 41:186–200, 1999. © 1999 Wiley‐Liss, Inc.     

Mating in Heliothis virescens: Transfer of juvenile hormone during copulation by male to female and stimulation of biosynthesis of endogenous juvenile hormone

Studies were undertaken to determine whether adult males of Heliothis virescens transfer juvenile hormone (JH) to females during copulation, and an in vitro radiochemical assay was used to determine whether mating causes an allatotropic effect, i.e., stimulation of JH biosynthesis by corpora allata (CA). In vitro, CA from 3-day-old mated females synthesized and released approximately 2.5 times total JH as that of CA from comparably aged virgin females. Of the homologues, JH II exhibited significant increase in mated females; JH I also increased but not significantly. JH III remained similar to that of virgin females. This is the first demonstration of an allatotropic effect of mating in moths. In contrast to the female, CA of virgin males did not produce any JH, but accessory sex glands (ASG) in 3-day-old males synthesized small amounts of JH. Immediately after adult emergence, male ASG contained approximately 1.5 ng JH I and II, which increased by 12 h after emergence and remained at this high level up to 54 h after emergence. JH III was barely detected in ASG. JH in ASG of mated male immediately after uncoupling was depleted almost completely, and 24 h later recovered to levels comparable to that of 54-h-old virgin male. Virgin female bursa copulatrix did not contain any JH, but mated female bursa, immediately after uncoupling, had JH at levels comparable to that observed in virgin male ASG. By 6 h after uncoupling, JH levels decreased dramatically in mated female bursa. These data suggest the transfer of JH to females by the male. Arch. Insect Biochem. Physiol. 38:100–107, 1998. © 1998 Wiley-Liss, Inc.     

Mate Choice by Non-Virgin Females Contributes to Reproductive Isolation between Populations of the Harlequin Beetle-Riding Pseudoscorpion

Determining whether reproductive isolation evolves through mate choice and/or gametic factors that prevent fertilization or through the post‐zygotic mechanisms of hybrid sterility or inviability is fundamental to understanding speciation. Investigation of the pre‐ and post‐zygotic components of reproductive isolation is facilitated in the pseudoscorpion, Cordylochernes scorpioides, by its indirect method of sperm transfer and viviparous embryonic development. Previous research on this species, in which mate discrimination was assessed in virgin females, suggested that female choice played only a minor role in reproductive isolation between populations from French Guiana and Panamá. Here, in a study of three allopatric populations of C. scorpioides from Panamá, we assessed mating‐stage isolation in both virgin and once‐mated females, and found that female discrimination depends critically on mating status. Virgin females were almost invariably receptive, showing no tendency to discriminate against males from allopatric populations. By contrast, non‐virgin females were significantly more likely to reject foreign males than males from their own population. Male sexual motivation could not account for differences in either female sexual receptivity or male success in sperm transfer. Allopatric and sympatric males did not differ in number of spermatophores deposited as either a female’s first or second mate. Nonetheless, allopatric males achieved significantly lower sperm transfer success not only with choosy, non‐virgin females but also with virgin females. Given the lack of behavioral discrimination by virgin females, female receptivity was not the only factor influencing differences in sperm transfer success. Multivariate analysis of spermatophore morphology suggests that the higher failure rate of matings between allopatric males and virgin females resulted from population differences in sperm packet architecture. Overall, our findings indicate that assessment of discrimination against allopatric males that is limited to virgin females may seriously underestimate the contribution of female mate choice to reproductive isolation between populations.     

Activity of the corpora allata of adult female Leucophaea maderae: effects of mating and feeding

Juvenile hormone III biosynthesis by corpora allata of adult female Leucophaea maderae was measured by an in vitro radiochemical assay. In fed females, JH III synthesis increases more than 20-fold after mating to a peak of 55 pmol/pair/h on day 9 and then rapidly declines. This increase in JH III synthesis concomitant with rapid oocyte growth in mated females is not observed in virgin females. The corpora allata from starved, virgin females appear to be inactive. The addition of 150 microM 2E,6E-farnesol (a) JH III precursor) to the incubation medium stimulates the corpora allata from starved, virgin females less than the corpora allata from starved, mated females. Both feeding and mating are necessary for the expression of a normal cycle of JH III synthesis in this cockroach.     

Juvenile hormone titre and egg production in Tenebrio molitor infected by Hymenolepis diminuta: effect of male and/or female infection,male age and mating

Infection of Tenebrio molitor with Hymenolepis diminuta induces curtailment of female fertility. We examined ovulation and oviposition, and associated titres of juvenile hormone (JH), in relation to parasitism and mating. Oviposition was significantly increased in infected mated and virgin beetles by days 6 and 9 post-emergence. Ovulation was not changed by infection; by the end of the 18-day experiment, the total number of laid eggs was not significantly altered. On day 6, JH levels were significantly higher in virgin infected insects, compared to non-infected controls (236+/-37.7 and 107+/-9.62 pg/g wet weight). Oviposition increased after mating, but total eggs ovulated remained the same. JH levels were higher in mated females on days 12 and 18 post-emergence, for infected and control insects. Previous studies suggested that male reproductive potential might rise following infection, because uninfected females lay more eggs when mated to infected males. We tested whether this caused an increase in female JH. Males were mated on days 5 or 12, when significant changes in their reproductive physiology begin to be observed, and are maximal, respectively. However, male age was of greater significance in promoting JH levels in females (p=0.001), than infection status of either partner (p=0.33).     

Rates of Juvenile Hormone biosynthesis and degradation during reproductive development and diapause in the boll weevil, Anthonomus grandis

Abstract. The role of Juvenile Hormone (JH) during reproductive development and diapause was investigated in the boll weevil, Anthonomus grandis Boheman (Coleoptera: Curculionidae). JH sythesized by corpora allata (CA) in vitro of A.grandis was identified as JH-UJ by high-performance liquid chromatography and by conversion to the methoxyhydrin. Optimal conditions for the use of a short-term assay in vitro were established to examine profiles of CA activity. In addition, rates of JH degradation by JH-specific esterase were determined. Patterns of CA and JH-esterase activity during reproductive development and the diapause state were established with laboratory-reared reproductive weevils and diapausing weevils collected as larvae and pupae in the field after the cotton-growing season. The results indicate that JH production is elevated in reproductive females whereas males and winter field-collected females show no CA activity. Vitellogenin concentrations in haemolymph and rates of oviposition were studied in relation to CA activity and JH degradation. An attempt to induce diapause in the laboratory failed.     

Developmental Role of Juvenile Hormone Metabolism in Lepidoptera

SYNOPSIS. Lepidopteran juvenile hormone (JH) esterase appearsto have a functional role in the regulation of embryogenesis,larval growth and development, and adult reproduction. In preovipositionaland newly laid eggs of the tobacco hornworm, Manduca sexta,JH esterase activity was elevated presumably to metabolize maternalJHs, and then declined after blastoderm formation. Also, a singlepeak in hemolymph JH esterase activity was found prior to ecdysisin the second through the fourth instar of M. sexta, the functionof which is unclear. However, in the last instar, elevated hemolymphJH esterase activity was noted prior to wandering and againprior to ecdysis to scavenge the last traces of JH necessaryfor normal development. The hemolymph JH esterase is likelyof multiple tissue origin for the prewandering peak with thefat body excluded as a source for the prepupal peak; an inhibitoryfactor from the brain and JH regulate JH esterase biosynthesis.In adult cabbage loopers, Trichoplusia ni, elevated hemolymphJH esterase activity appeared to be important in reducing theJH titer and preventing egg maturation. Structure/activity datawith trifluoromethylketones were incorporated into the designof a novel, JH esterase inhibitor, the sulfone and hydrate ofoctylthio-1,1,1- trifluoropropan-2-one, with selective and persistent,in vivo inhibitory activity. The topical application of thiscompound to last instar larvae and virgin adults of T. ni producedjuvenizing effects (delayed pupation and induced egg maturation/oviposition,respectively) providing direct evidence of a functional rolefor JH esterase in lepidopteran development.     

Mating Behavior and Dual‐Purpose Armaments in a Camel Cricket

Sexual selection has often been dichotomized into intersexual and intrasexual components of selection, favoring ornaments and weapons, respectively. Here, we show that a weapon used in male–male combat is sometimes also used in a functionally similar manner for grasping females during mating. The hind legs of adult males of some species of Pristoceuthophilus camel crickets have strongly bent tibia and stout femora with two large conspicuous spines. Here, we show that (1) leg armature is positively allometric, (2) males use these leg modifications when fighting other males, (3) males sometimes use the same leg modifications in the same functional manner for grabbing and holding females for mating, (4) virgin females show more interest in males than do non‐virgin females, and (5) males are more likely to use their leg modifications to grab and hold females when paired with non‐virgin females than when paired with virgin females. We interpret these results as suggesting that males employ an alternative coercive mating strategy when paired with unreceptive females; alternative explanations are also discussed.     

Hormonal control of egg maturation in the corn earworm, Heliothis zea

At eclosion, the ovaries of female Corn earworm Heliothis zea do not contain mature eggs. Virgin-unfed females produced approximately 400 mature eggs in 8 days; mating or feeding doubled this number, and mating plus feeding more than tripled it. Females allatectomized or decapitated at day O matured few eggs. Egg production was restored by implantation of active corpora allata (CA) or by treatment with the juvenile hormone (JH) analogue methoprene at day 0. 20-Hydroxyecdysone, on the other hand, had no effect. Females in which the CA had been denervated or in which the median neurosecretory cells of the brain had been ablated at day O produced fewer eggs than sham-operated animals. These results indicate that egg maturation is controlled by JH and that continuous input from the brain is required for sustained CA activity for maintaining a high rates of egg maturation.The rate of JH biosynthesis by CA in vitro was determined with a radiochemical assay. The major hormones produced were JH-II and JH-III with small quantities of JH-I. The rates of JH synthesis were similar in all experimental groups which may indicate that the in vitro rate of JH synthesis does not reflect the actual state of CA activity in the female.     

Morph-specific diurnal variation in allatostatin immunostaining in the corpora allata of Gryllus firmus: Implications for the regulation of a morph-specific circadian rhythm for JH biosynthetic rate

Previous studies have documented a circadian cycle in juvenile hormone (JH) biosynthesis in the long-winged, flight-capable morph, but not in the short-winged flightless morph of the cricket Gryllus firmus. One rapid and reversible inhibitor of in vitro JH biosynthesis by the corpora allata (CA) in crickets is the neuropeptide Phe-Gly-Leu/Ile-amide type of allatostatins (ASTs). To investigate the possible role of allatostatin regulation of the morph-specific circadian cycle of JH production, the quantity of this type of AST in the nerves within the CA was determined by the density of anti-AST-immunostaining in confocal images using the Image J program. The density of immunostaining was inversely related to the rate of JH biosynthesis: Immunostaining in the CA was high and did not differ between morphs early in the photophase when the in vitro rate of JH biosynthesis is low and equivalent in the morphs. However, during the end of the photophase, when the rate of JH biosynthesis rises dramatically in the flight-capable morph, but not in the flightless morph, immunostaining was significantly lower in the flight-capable compared to the flightless morph. These results indicate that morph-specific differences in delivery of AST to the CA and its probable release likely regulate the morph-specific circadian pattern of JH biosynthesis. Also, the negative correlation between AST density and JH production provides evidence for predicting the periods of altered release of these rapid-acting paracine regulators of JH biosynthesis.     

Activity of the corpora allata of adult female Aedes aegypti: effects of mating and feeding

The synthesis of juvenile hormone III (JH III) by the isolated corpora allata (CA) of Aedes aegypti adult female was studied using an in vitro radiochemical assay. We dissected the corpora allata-corpora cardiaca (CA-CC) complex attached to a piece of aorta. The complex was left connected to the intact head capsule to facilitate the visualization and transfer of the glands. A linear increase in the cumulative amount of biosynthesized JH III was found for at least the first 6 h of incubation; approximately 45% of the synthesized JH III was present in the medium. There was a dependence of JH III synthesis on exogenous methionine supply. Using reversed phase high performance liquid chromatography two major labeled products biosynthesized by the CA were separated. They co-migrated with JH III and methyl farnesoate (MF). The identity of the biosynthesized JH III was confirmed by gas chromatography-mass spectrometry. JH III synthesis was only 2.0 fmol/pair gland/h immediately after adult emergence, but increased to 32.6 fmol/ pair gland/h 18 h later in sugar-fed females. Two days after emergence, the CA biosynthetic activity slowly started to decrease, and reached values of around 5.3 fmol/pair gland/h by one week after emergence. Synthesis of JH was similar from either sugar-fed females mated or unmated. A blood meal resulted in a decrease of JH III synthesis in CA from mated females by 12 h after feeding and from virgin females by 24 h after feeding. JH III biosynthesis remained low for at least 96 h in mated females, but was back to higher levels 72 h after feeding in virgin females. Rates of JH III biosynthesis closely reflected the hemolymph levels of JH III both after emergence and after a blood meal described by Shapiro et al. (1986). The activity of the CA in Aedes aegypti females seems to be regulated by developmental changes and nutritional signals, and to be independent of mating stimulus.     

Juvenile hormone biosynthesis and oocyte development in adult female Blattella germanica: Effects of grouping and mating

The roles of grouping and mating in modulating the activity of the corpora allata (CA) in adult female cockroaches were investigated using the in vitro radiochemical assay of juvenile hormone (JH) biosynthesis. Isolated virgin females have longer, asynchronous cycles of CA activity and oocyte maturation than do isolated females mated on day 8. Three factors were identified as the major contributors to this difference: (1) an experimental artifact of selection for sexually receptive females, (2) a positive effect of grouping on JH synthesis and oocyte maturation, and (3) a positive effect of copulation on oviposition and retention of the ootheca. Mated females constitute a subpopulation of receptive females that differ significantly from other females by having higher rates of JH synthesis prior to mating. The relative importance of such selection is substantial when the rate of mating is low, as in experiments with isolated females that are exposed to males for a short period of time. Long-term exposure of females to males introduces a grouping effect, which obscures any additional effect of mating on CA activity and oocyte development. However, mating influences ootheca formation and its retention. The effect of grouping can be mimicked in isolated females by transection of the nerves connecting the CA–corpora cardiaca complex to the brain, suggesting that in this insect isolation causes brain inhibition of the CA, and grouping provides disinhibitory stimuli that release the CA from brain inhibition.