Degradation of juvenile hormone and methylation of juvenile hormone acid by corpora cardiaca–corpora allata of the cockroach,Nauphoeta cinerea: I. Biochemical aspects

Corpora cardiaca-corpora allata (CC-CA) from vitellogenic females of Nauphoeta cinerea degraded, in vitro, racemic and (10R)-juvenile hormone III (JH III) at a rate of 249 pmol/CC-CA/h and 786 pmol/CC-CA/h, respectively. The major metabolite formed was JH III acid, together with some highly polar products. CC-CA homogenates degraded racemic JH III to a small extent, whereas (10R)-JH III was degraded efficiently to JH III acid. No highly polar products were formed by CC-CA homogenates. When CC-CA were incubated with racemic JH III acid, some of this substance was degraded to highly polar products, and a minor part was methylated to JH III. CC degraded very little JH III acid and did not methylate it to JH III. CC-CA homogenates methylated JH III acid very efficiently; we measured an apparent K_max of 37.8 μM and a V_max of 1,260 pmol/4 h/ CC-CA equivalent. The addition of JH III acid to CC-CA in vitro increased the rate of biosynthesis of JH III, as determined by measuring incorporation of methyl[¹⁴C]methionine into JH III. These data indicate that the metabolite JH III acid can enter the CA and be methylated to JH III.     

Conditions for estimation of corpus allatum activity in the blowfly, Phormia regina, in vitro

ABSTRACT. Incubation conditions have been established for the corpus cardiacum-corpus allatum (CC-CA) complex of female Phormia regina (Meigen), which will support CC-CA biosynthetic activities in vitro as measured by the incorporation of a labelled methyl group with L-[methyl-³H]methionine as the methyl donor. After incubation, radioactivity in the organic extract of the medium was determined by scintillation counting. Analysis of the organic extract with reverse phase high-performance liquid chromatography (HPLC) revealed that a compound which has similar retention time (UV absorbance) with synthetic JH III was synthesized by the CC-CA complexes of liver-fed females. By using this short-term, in vitro , radiochemical assay for CA activity, it was shown that a protein diet significantly increases the activity of the CA compared with females fed only a sugar-water diet. Furthermore, use of HPLC separation, in conjunction with scintillation counting of time-collected fractions, demonstrated the existence of a moiety containing incorporated radiolabeled methyl group (from the methionine) which did not co-elute with JH I or JH III standards. These results suggest that in P. regina use of the incorporation of a radiolabeled methyl group to measure JH biosynthesis (CA activity) can be misleading if the compounds which do not co-elute with JHs are not considered.     

Apparent inhibition of in vitro juvenile hormone biosynthesis by the corpus cardiacum of adult crickets (Gryllus bimaculatus and Acheta domesticus) is due to juvenile hormone esterase

When measuring the in vitro JH III-biosynthesis by corpora allata (CA) from adult female crickets in the presence of corpora cardiaca (CC), the amount of JH III in the medium decreased in a dose dependent manner. The CC of a 4-day-old female Gryllus bimaculatus contain 42 pmol.pair CC⁻¹ Grb-AKH, 0.62 pmol.pair CC⁻¹ octopamine, and a JH-esterase activity of 9.8 pmol JH.h⁻¹.pair CC⁻¹. Comparable values for Acheta domesticus are 21 pmol.pair CC⁻¹ Grb-AKH, 0.53 pmol.pair CC⁻¹ octopamine, and 6.5 pmolJH.h⁻¹.pair CC⁻¹ of JH-esterase activity. Even if the entire octopamine content of the CC were released into the medium, the concentration would be below the 10⁻⁵ M threshold for octopamine inhibition of JH synthesis. An in vitro AKH inhibition of JH III synthesis was observed, but only at a relatively high concentration (10⁻⁵ M). If the entire AKH content (10⁻⁶ M) of the CC were released into the medium, the AKH concentration would approach JH synthesis inhibiting levels. However, the rate of release of AKH in vitro was very low, and, therefore, AKH from the CC could not affect JH synthesis. In contrast, a specific JH-esterase, released by isolated CC into the medium, was sufficiently high in both cricket species to account for the observed decrease in JH III present. OTFP-sulfone (10⁻⁵ M) restored apparent JH synthesis of the CA to the control level. There was no reduction in the amount of JH released when CA were incubated with heat treated CC. The CA themselves contained almost no JH-esterase activity. © 1997 Wiley-Liss, Inc.     

Biosynthesis and regulation of juvenile hormone III,juvenile hormone III bisepoxide,and methyl farnesoate during the reproductive cycle of the grey fleshfly,Neobellieria (Sarcophaga) bullata

To study the effect of brain signals on the biosynthesis of juvenile hormone by the corpora allata of the grey fleshfly Neobellieria bullata, exposed corpora allata connected to the brain were surgically removed from sugar-fed flies and incubated in vitro with L -[³H-methyl]methionine. After incubation, the media together with the tissues were analyzed by HPLC. [³H]Juvenile hormone III (JH III), [³H]JH III bisepoxide (BE), [³H]methyl farnesoate (MF) and an unknown [³H]labeled metabolite (Un) were identified as the primary products. The rate of synthesis of [³H]JH III bisepoxide was higher than that of [³H]JH III, [³H]MF and [³H]Un. Two days after a liver meal, female flies synthesized more JH III, MF, BE, and the Un than did males. Synthesis of JH III, BE, and MF in females was lower during the previtellogenic, sugar-feeding period than during the vitellogenic liver-feeding period. Isolated corpus cardiacum–corpus allatum (CC-CA) complexes that were incubated in vitro synthesized less JH III, MF, and BE, as compared to complexes that were attached to the brain, indicating that the brain probably modulates the biosynthesis of JH III, MF, and BE in the corpora allata. Upon incubation of brain–CC–CA complexes with Neb-TMOF (10^–8 M), Neb-colloostatin (10^–8 M), ovarian, or brain extracts resulted in significant inhibition of JH III and BE biosynthesis in the presence of ovarian extracts. These results indicate that allatostatin-like factors are present in the ovary of the flesh fly. Arch. Insect Biochem. Physiol. 37:248–256, 1998. © 1998 Wiley–Liss, Inc.     

Juvenile hormone biosynthesis by corpus cardiacum-corpus allatum complexes of larval Lymantria dispar

• 1.1. A radiochemical assay was used to examine juvenile hormone (JH) synthesis and secretion in vitro by incubating two pairs of larval corpus cardiacum-corpus allatum complexes (CC-CA) from, Lymantria dispar, in 50 μl of osmotically balanced Grace's medium containing 1 μC1 [³H-methyl]-methionine for 6 hr.
• 2.2. For CC-CA of fourth instar female larvae, maximal incorporation of ³H-methyl was 0.15 pmol/pr/hr between days 2 and 3. High pressure liquid chromatographic (HPLC) analysis suggested that the biosynthetic products are mainly JH III with a little JH II at times.
• 3.3. For CC-CA of last instar female larvae, incorporation of ³H-methyl was 0.48 pmol/pr/hr at the beginning of the stadium and decreased to negligible levels by day 10. HPLC analysis suggested that CC-CA of last instar larvae produced only JH III. Volume increases in CA during the last instar were associated with declining activities of JH secretion.
• 4.4. Comparisons of maximal rates of ³ H-methyl incorporation by each unit volume of CA revealed that in the last instar each unit volume (μm³) of glandular tissue secreted 50% more JH than in the fourth instar.

     

Blood meal as a regulator of triacylglycerol synthesis in the haematophagous stable fly,Stomoxys calcitrans

Summary Accumulation of triacylglycerol and glycogen reserves following meals of blood and/or sugar-water was investigated in the haematophagous stable fly,Stomoxys calcitrans. In the starved fly, triacylglycerol reserves appear to be utilized predominantly for energy requirements. When the starved flies were fed one meal of 1.0 M sucrose-water, they accumulated considerable amounts of glycogen but there was no increase in the triacylglycerol content. Starved flies fed one meal of blood accumulated large quantities of triacylglycerol but no glycogen; in those flies fed sugar-water and blood, glycogen and triacylglycerol accumulated. This study shows that the stable fly preferentially utilizes blood for triacylglycerol synthesis and sugar for glycogen synthesis. It is suggested that one or more factor(s) in the blood meal influences accumulation of triacylglycerol in this insect, possibly through an hormone from the corpora cardiaca-corpora allata complex.     

Degradation of juvenile hormone and methylation of juvenile hormone acid by corpora cardiaca–corpora allata of the cockroach,Nauphoeta cinerea: II. Physiological aspects

In vitellogenic females of Nauphoeta cinerea, injected (10R)-juvenile hormone (JH) III was degraded more rapidly than racemic JH III: we measured a half-life of 21 min (with or without coinjection of lipophorin) for the former and 24 min (with coinjection of lipophorin) and 43 min (without coinjection of lipophorin) for the latter. One to two hours after injection, JH III acid was the major metabolite observed; in addition, several highly polar products were found. The half-life of injected racemic JH III acid was 19 min with coinjection of lipophorin and 4 min without. The JH III acid titer in hemolymph was low (around 5–10 pmol/ml) in last instar larvae and previtellogenic and pregnant females and reached higher values (40–100 pmol/ml) in vitellogenic and ovulating females. Racemic JH III acid could be methylated in vitro to JH III by corpora cardiaca–corpora allata (CC-CA) from penultimate instar larvae and females at stages between adult ecdysis and ovulation and at the very end of pregnancy, but not by CC-CA from last instar larvae and adult females at earlier stages of pregnancy. This indicates that CC-CA are capable of methylating JH III acid only at stages when JH III is detectable in the hemolymph. In double-labelling experiments with CC-CA from vitellogenic females and L-[¹⁴C]methionine and [³H]JH III acid as precursors, we observed that only a small proportion (1–8%) of total biosynthesized JH III was derived from JH III acid when the latter was present at physiological concentration. This suggests that in vivo recycling of JH III acid by CC-CA plays only a minor role in the regulation of the titer of JH III and JH III acid.     

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.     

Spatial expression of the mevalonate enzymes involved in juvenile hormone biosynthesis in the corpora allata in Bombyx mori

The developmental expressions of the mRNA of JH synthetic enzymes have been studied using homogenates of the corpora cardiaca-corpora allata (CC-CA) complexes in Bombyx mori [Kinjoh, T., Kaneko, Y., Itoyama, K., Mita, K., Hiruma, K., Shinoda, T., 2007. Control of juvenile hormone biosynthesis in Bombyx mori: cloning of the enzymes in the mevalonate pathway and assessment of their developmental expression in the corpora allata. Insect Biochemistry and Molecular Biology 37, 808-818]. The in situ hybridization analyses in the CC-CA complex showed that the distribution of the mRNAs of all the mevalonate enzymes and juvenile hormone (JH) acid O-methyltransferase occurred only in the CA cells, indicating that the fluctuations of the enzyme mRNA amounts in the CC-CA complexes were derived solely from the CA. In addition, the size of the CA and their nuclei was not associated with the JH synthetic activity by the CA until the pharate adult. Only female adult CA synthesized JH in B. mori, and the CA and the nuclei were significantly larger than those of male CA which do not synthesize JH.     

Hydrolytic degradation of juvenile hormones in haemolymph and corpora allata of Galleria mellonella (L).

Hydrolytic rates of juvenile hormones (JHs) I, II and III by the corpora cardiaca-corpora allata complex (CC-CA) and by the haemolymph of Galleria mellonella remain in the same order (III greater than I greater than II in CC-CA and I greater than III greater than II in haemolymph) throughout the last larval instar. Haemolymph hydrolytic activity shows peak at the end of feeding when 80 pmol JH I versus 15 pmol JH II is degraded per 1 microliter and minute; hydrolysis rapidly declines in the apolysing insects. Hydrolytic rates in CC-CA reach a maximum of 240 fmol/pair per min for JH III and 85 fmol/pair per min for JH II in pharate pupae. Brain implantations or chilling of freshly ecdysed last instar larvae, which are known to elevate JH titer and induce supernumerary larval molt, do not affect JH hydrolysis. The results indicate that the dominance of JH II in Galleria may be at least partly controlled by preferential hydrolysis of homologs I and III.     

Juvenile hormone biosynthesis in the fall armyworm, Spodoptera frugiperda (Lepidoptera, Noctuidae)

The in vitro production of juvenile hormones (JH) was investigated by using corpora allata (CA) of larvae and corpora cardiaca-corpora allata (CC-CA) complexes of adult females of the fall armyworm Spodoptera frugiperda. In female moths, JH release was high compared to that in 5th and 6th instar larvae. Concentrations of 0.11-0.12 mM methionine, 180-200 mM Na(+), 5.8-8.3 mM K(+), 10-50 mM Ca(2+) and a pH range of 5.7-6.3 yielded optimal incorporation of L-[methyl-(3)H] methionine in vitro by CC-CA complexes. The highest hourly incorporation occurred during a 9-h incubation period following a 1.5-h lag-phase. JH release from CC-CA complexes of adult females was shown to be age-dependent with a peak value on day 2 (approx. 4 pmol h(-1) CA(-1)). By a combination of reversed phase (RP)- and normal phase (NP)-high performance liquid chromatography (HPLC), two major labelled products released by the complex were separated. One compound co-migrated with chemically synthesized JH II diol, the second compound with JH III diol. Only traces of JH II and III could be detected in some samples. Gland extracts also contained both the major radiolabelled products. Double labelling experiments using [3H]methionine and [14C]acetate confirmed their de novo synthesis in CC-CA complexes of female moths. The nature of chemically synthesized reference JH III diol was proved by LC-MS (ESI mass spectrometry) and 1H-NMR (nuclear magnetic resonance spectroscopy).     

Allatotropic activity in the brain of female Phormia regina (Diptera: Calliphoridae)

In Phormia regina, the rate of juvenile hormone (JH) synthesis rises rapidly after the ingestion of an adequate protein meal. In a previous publication we have localized the neurons containing Manduca sexta allatotropin (Mas-AT)-like substances in the brain of P. regina and demonstrated the allatotropic effect of synthetic Mas-AT in sugar-fed flies in vitro. In this current study, we examined the possible role of the brain of P. regina after the fly received a protein meal. In vitro studies showed that the brain releases, at 8 h after a protein meal, a factor(s) with a strong allatotropic effect. This factor(s) stimulates the corpus allatum (CA) to produce 6.9 times more juvenile hormones (JHs) than the control CA. Time course studies showed that the release of this allatotropic factor(s) is temporally controlled. Only the brains collected from flies at 6 and 8 h after the onset of a liver meal release allatotropic factor(s). Injection of anti-Mas-AT antiserum partially suppressed the fly follicle development in vivo. Presence of anti-Mas-AT antiserum decreased the allatotropic effect of the brain released allatotropic factor(s) in vitro. In addition to a Mas-AT-like substance, it is possible that the brains of liver-fed P. regina females may synthesize other allatotropic factors that are chemically unrelated or partially related to Mas-AT, which cannot be recognized/neutralized by our anti-Mas-AT antiserum.     

家蝇卵巢摄取卵黄蛋白的机理

在家蝇Musca domestica viaina 的卵黄发生过程中,卵母细胞摄取卵黄原蛋白与滤泡开放是相关的。观察不同发育时期的家蝇滤泡结果表明,在摄取活动最旺盛的时期也就是卵黄发生的顶盛时期,其滤泡开放程度最大,而在卵黄发生前期和后期基本上没有摄取活动,此时的滤泡上皮细胞间不开放。卵巢体外培养的激素处理表明,JH可以促进滤泡开放。家蝇卵巢微粒体制备物的Na⁺-K⁺ATP酶活力在卵巢发育过程中存在着动态变化。羽化后24小时时有一定的酶活性,随着卵黄发生的进行,酶活力逐渐增加,到羽化48小时时酶活力最高,然后又开始下降,到羽化72小时时已经很小。羽化32小时的家蝇点滴或注躬 JH之后,测得的卵巢微粒体制备物的Na⁺-K⁺ATP酶活力比正常羽化36小时的高,羽化44小时的家蝇点滴和注射JH之后,测得酶活力比正常羽化48小时的低。羽化36小时和48小时的家蝇卵巢微粒体制备物与JH共同作用后,其Na⁺-K⁺ATP酶的活力分别增加2.95倍和3.50倍,羽化48小时的家蝇卵巢在含有JH的培养液中培养启,其匀浆液的酶活性为对照组的1.26倍。 由此我们可以推测在家蝇的卵黄发生过程中,JH通过促进滤泡开放和增加卵巢微粒体制备物Na⁺-K⁺ATP酶的活力,从而调控卵母细胞对卵黄蛋白的摄取。     

A putative juvenile hormone in a stink bug,Plautia stali: The corpus allatum produces and releases a JH-active product different from any known JHs in vitro

Summary

A radiochemical method was adopted to analyze the in vitro products of the corpus allatum (CA) of Plautia stali. The radiolabel derived from ³H-methionine added to the incubation medium was incorporated and released by CA as two main radiolabelled products. They showed Rf values of about 0.3 and 0.5, respectively, in the thin layer chromatography (TLC) system used. The release of these products was shown to be CA-specific since in control incubations using the brain, midgut, aorta and flight muscle, virtually no release of these products was observed. The locations where these main products migrated on the TLC did not coincide with spots of synthetic standards of JH I-III or JHB₃, a JH found in higher Diptera. An addition of precursors of JH III, farnesoic acid or farnesol stimulated the CA to biosynthesize the products with an Rf value of 0.5 up to about 10-fold, suggesting that the product in question may have a sesquiterpenoid skeleton similar to JH III. Topical applications of the hexane extracts of the medium in which the CA had been incubated exerted a juvenilizing, metamorphosis-inhibiting effect on final instar nymphs in a dose-dependent fashion. The nymphs treated with the hexane extracts at a high dose moulted to intermediates with reduced forewings and scutellum, as well as nymphs implanted with the CA from reproductively active females. Based on this juvenilizing effect found in the hexane extracts, the JH-active fraction was determined after TLC separation. This assay indicated that the products found at an Rf value of about 0.5 was JH-active. These results suggest the presence of a new JH different from any known JHs in P. stali.     

Allatotropic and allatostatic activities in extracts of brain and the effects of Manduca sexta allatotropin and Manduca sexta allatostatin on juvenile hormone synthesis in vitro by corpora allata from the Eri silkworm, Samia cynthia ricini

Both allatotropic and allatostatic activities were found in crude extracts of brain from adult and larval Eri silkworm, Samia cynthia ricini, but it seems that allatotropic activity dominates in each stage. There was a high level of allatotropic activity in the crude extract of brain from newly emerged female adults, but allatostatic activity appeared in the bioassay when excessive amounts of crude extracts of brain were added. Crude extracts of brain from premoulting fourth‐instar larvae and from newly ecdysed fifth‐instar larvae exhibited allatotropic activities, whereas extracts of brain from the second and third day of the fifth‐instar larvae inhibited juvenile hormone (JH) release slightly. Allatotropic activity from the brains of adults and larvae stimulated both adult and larval corpora allata (CA) to synthesize JH. Manduca sexta allatotropin (AT) (Mas‐AT) and M. sexta allatostatin (AST) (Mas‐AST) also stimulated and inhibited both adult and larval S. cynthia ricini CA to synthesize JH, respectively. Higher concentrations of Mas‐AT (10^?4 or 10^?3 M) showed an inhibitory effect on adult CA. CA from newly emerged female adults were the most sensitive to inhibition by Mas‐AST, whereas CA from female pharate adults at about 6 h before adult emergence were the most sensitive to stimulation by Mas‐AT and S. cynthia ricini brain allatotropic activity. An extract of brain and Mas‐AT induced some of the non‐active female pharate adult CA at 12 h before emergence to synthesize a small amount of JH.     

Biosynthesis of (10R)-Juvenile hormone III from farnesoic acid by Aedes aegypti ovary

Synthesis of (10R)-juvenile hormone III (JH III) outside the corpora allata (CA) was investigated in female Aedes aegypti. Intact females or ligated abdomens of blood-fed and sugar-fed females synthesized in vivo [12-³H]JH III-like molecules from [12-³H]-methyl farnesoate, indicating that an organ(s) in the female abdomen, other than the CA, converted methyl farnesoate into JH III. To find out the organ(s) that synthesized JH III-like molecules, ovaries, fat bodies, and midguts were incubated in vitro with [12-³H]methyl farnesoate and the synthesis of JH III-like molecules was compared with JH III synthesized by CA. To identify tissue(s) having both farnesoic acid methyl transferase and farnesoate epoxidase, enzymes that convert farnesoic acid into JH III, ovaries, and fat bodies were removed from sugar and blood-fed females and incubated with [12-³H]farnesoic acid. Chemical derivatization by methoxyhydrin formation followed by esterification with (+)-α-methoxy- α-trifluoromethyl phenylacetic (MTPA) acid chloride and reversed phase liquid chromatography identified (10R)-JH III methoxyhydrin (+)-MTPA ester as the sole JH III-like molecule produced in tissue culture incubation of ovaries. Since only (10R)-JH III is produced and not racemic JH III, the oxidation of farnesoic acid must be enzymatically mediated. Ovaries and corpora allata of female A. aegypti also synthesized [³H,¹⁴C]JH III from L-[methyl-³H]methionine and [¹⁴C]acetate which was characterized by HPLC and gas chromatography. These results suggest that mosquito ovary can synthesize (10R)-JH III from farnesoic acid, and that this tissue synthesizes JH III-like molecules from L-methionine and acetate. © 1994 Wiley-Liss, Inc.     

Comparative metabolism of isoleucine by corpora allata of nonlepidopteran insects versus lepidopteran insects,in relation to juvenile hormone biosynthesis

We studied the metabolism of [U-¹⁴C]isoleucine by intact and homogenized corpora allata (CA) from various insect species to determine how this substrate is converted to precursors of juvenile hormone (JH). CA homogenates of the lepidopterans Manduca sexta, Hyalophora cecropia, and Samia cynthia metabolize [U-¹⁴C]isoleucine to several products including 2-keto-3-methyl-valerate, 2-methylbutyrate, CO₂, propionate, and acetate. Intact CA of male H. cecropia produce particularly high levels of 2-keto-3-methylvalerate, indicating a highly active branched-chain-amino acid transaminase. In contrast, CA homogenates from the nonlepidopterans Periplaneta americana, Schistocerca nitens, Tenebrio molitor, and Diploptera punctata barely metabolize [U-¹⁴C]isoleucine. However, P. americana CA homogenate metabolizes [U-¹⁴C]2-keto-3-methylvalerate, the transamination product of [U-¹⁴C]isoleucine, more rapidly than does a homogenate of M. sexta CA. Furthermore, intact CA from P. americana incubated with [U-¹⁴C]2-keto-3-methylvalerate incorporate low levels of ¹⁴C into JH III, but do not metabolize this substrate to JH II or JH I. Intact CA from female Diploptera punctata produce very high levels of JH III, but are also unable to incorporate radiolabel from [U-¹⁴C]isoleucine into JH III, which substantiates our findings with other nonlepidopteran CA. The results suggest that CA of nonlepidopteran insects lack an active branched-chain amino acid transaminase and, consequently, are unable to utilize these substrates for JH biosynthesis.     

Oosorption in the stink bug Plautia stali: role of Juvenile hormone in the induction of oosorption

To clarify the role of Juvenile hormone (JH) in the induction of oosorption in females of the stink bug Plautia stali Scott (Heteroptera: Pentatomidae), the effects of extirpation and implantation of the corpus cardiac‐corpus allatum complex (CC‐CA) are examined in fed and starved adults, respectively. Removal of CC‐CA induces oosorption in fed females, whereas CC‐CA implantation stimulates ovary development in starved females. Transection of the nervous connections between the brain and CC‐CA also exerts a stimulatory effect on ovary development. Uptake of yolk protein by the oocytes, assessed in terms of incorporation of a fluorescence dye, occurs on the day after food deprivation but ceases within 1 day after allatectomy. When females are deprived of food, beginning on day 3 of adult life, and treated with JH III skipped bisepoxide (JHSB₃) on the same day, their ovaries develop in a dose‐dependent fashion. Approximately half of the starved females that received JHSB₃ application on day 5 undergo oosorption in the terminal oocytes. This indicates that the critical starvation period for oosorption induction is approximately 2 days, and the earlier half of this period may reflect the time required for the brain to detect poor nutritional condition. During the latter half, in response to food deprivation, the brain inhibits JH biosynthesis by the corpus allatum through nervous connections, resulting in a low JH titre, which in turn induces oosorption.     

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.     

Changes in the size of corpora allata,in the juvenile hormone III titer in the hemolymph,and in the protein content of terminal oocytes throughout the first gonadotropic cycle in Aiolopus thalassinus (Insecta: Orthoptera: Acrididae)

In Aiolopus thalassinus (Fabr.), the changes in the volume of the corpora allata (CA), in the concentration of juvenile hormone III (JH III) in the hemolymph, and in protein content in the terminal (t) oocytes were studied during the first gonadotropic cycle. These parameters could be better related to the volume of t-oocytes than to age after emergence. The JH III titer curve was maximum (2.9 pmol/10 μl) at an oocyte volume of 1.2 mm³. Before oviposition (days 10–16) the JH III titer decreased to 0.65 pmol/10 μl hemolyph. The increase in JH III titer reflected a period of high protein storage in the t-oocytes. The largest volume of the CA was reached at the beginning of yolk storage in the t-oocytes. The highest JH III titer did not correspond with the largest volume of CA, which occurred much earlier. © 1993 Wiley-Liss, Inc.