Change in corpus allatum function during metamorphosis of the tobacco hornworm Manduca sexta: Regulation at the terminal step in juvenile hormone biosynthesis

Changes in activity of the corpora allata (CA) during larval-pupal-adult development of the tobacco hornworm Manduca sexta were studied by transplantation assays, measurements of in vitro juvenile hormone (JH) and JH acid synthesis, and determination of JH acid methyltransferase (JHAMT) and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activities. The data from these assays demonstrate that the CA cease to secrete JH by day 4 of the last larval instar (wandering stage). With regard to JH synthesis, they remain inactive throughout the prepupal, pupal, and most of the pharate adult periods. CA of females, but not of males, resume JH synthesis shortly before eclosion. The biochemical basis of the inactivation process is the loss of JHAMT activity. However, prepupal CA produce JH acids, as shown by enzyme and in vitro assays. Pupal and pharate adult CA do not synthesize JH acids although levels of HMG-CoA reductase activity seem to remain relatively high. Radiolabeled JH was recovered from hemolymph of allatectomized prepupae that had been injected with radiolabeled JH acid. These results provide further evidence that certain peripheral tissues (eg, imaginal discs) convert JH acid secreted by the prepupal CA to JH and, thus, that JH acid is a prohormone in the prepupal period. The CA change from hormone secretion to prohormone secretion during larval-prepupal transformation, a unique functional alteration in an endocrine gland.     

Morphogenetic and gonadotrophic effects of a juvenile hormone analogue ((7S)-hydroprene) in last instar female larvae of Diploptera punctata

ABSTRACT. The effect of treatment of last instar female larvae of Diploptera punctata with a Juvenile Hormone (JH) analogue, (7 S )-hydroprene, has been determined with respect to the ability of the analogue to alter the duration of the stadium and the nature of the ensuing ecdysis. We have also investigated the effects of the analogue on JH release, the growth of the basal oocytes, as well as ecdysteroid titres during the fourth stadium. Analogue treatment prior to day 10 of the stadium results in prolongation of the stadium and desynchronization of ecdysteroid release. Thereafter, treatment with the analogue has little effect. Analogue treatment results also in the formation of supernumerary larvae and intermediates, in a dose-dependent fashion, provided that animals are treated on day 10 or earlier. Thus, the 'critical' period for metamorphosis in last instar D. punctata is between days 0 and 10.
Treatment with (7 S )-hydroprene produces profound effects also on both JH release, and basal oocyte growth. At a dose of 500μg administered on day 1, JH release is stimulated significantly at a time when JH release is normally undetectable. Significant growth of basal oocytes is observed in such treated animals, and appears to precede the peak in JH release. We suggest that the growth of the basal oocytes, as a result of analogue treatment, stimulates the production of JH by CA in these last instar larvae.     

Juvenile hormone and reproduction in crickets, Gryllus bimaculatus De Geer: corpus allatum activity (in vitro) in females during adult life cycle

ABSTRACT. Incubation conditions were established for a short-term radiochemical assay of spontaneous juvenile hormone (JH) biosynthesis in vitro by corpora allata from adult female Gryllus bimaculatus. The only JH synthesized was shown by HPLC to be JH III. A further incubation product, predominantly extracted from the corpora allata, was thought to be the JH III precursor, methyl farnesoate. In adult females reared at a constant temperature of 27°C the synthetic activities of the corpora allata-corpora cardiaca complexes in vitro increase from almost zero to a high peak value 4 days after the imaginal moult. Thereafter the activity decreases to varying intermediate levels, but always lower than the first maximum. Two days after the first peak in corpus allatum activity, ovarian fresh weight increases dramatically and the first oviposition occurs 2 days later.
Topical application of JH III to females reared at 20°C, which usually have a low fecundity, causes a dose-dependent stimulation of egg production and oviposition.     

Juvenile hormones: Their role in the regulation of the pheromonal communication system of the armyworm moth,Pseudaletia unipuncta

Recently, much effort has been devoted to the elucidation of the neuro-endocrine mechanisms regulating the biosynthesis and emission of sex pheromones in the Lepidoptera. The available data indicate that the hormonal mechanisms involved vary considerably among species. For example, compelling evidence that juvenile hormones (JH) play a role in the control of sex pheromone production has been presented only for the armyworm moth, Pseudaletia unipuncta. In this species, females that are allatectomized at emergence neither produce nor release pheromone, but both activities are restored following replacement therapy with synthetic JH. However, injection of synthetic JH into neck-ligated females does not induce pheromone biosynthesis, whereas treatment with either a brain homogenate or synthetic PBAN results in a rise in the pheromone titer. These results indicate that the role played by JH is an indirect one and that the tropic factor is a PBAN-like substance. Studies on in vitro JH biosynthesis by isolated corpora allata of P. unipuncta have shown that the low JH output observed early in the life of adult females coincides with the absence of both calling behavior and pheromone production. The subsequent increase in the rates of JH biosynthesis correlates with the onset of pheromone production and release. We have therefore proposed that JH titers must pass a threshold level before the circadian release of PBAN and calling behavior can begin. Furthermore, recent experiments suggest that the continuous presence of JH is necessary for calling behavior to be maintained once initiated. Lastly, we present data suggesting a role for JH or JH acids in the receptivity of P. unipuncta males to the female sex pheromone. © 1994 Wiley-Liss, Inc.     

Variability in Juvenile Hormone production by locust corpora allata kept in vitro for long periods

Abstract. Juvenile Hormone III (JH-III) production by corpora allata (CA) of sexually mature female locusts (Locusta migratoria migratorioides (R. & F.)) was maintained in vitro for up to 30 days in an agar-solidified medium. Hormone production was measured periodically with a short-term radiochemical assay. Low-activity CA increased their activity significantly after 24–48 h incubation in the long-term medium, but high-activity glands did not. Variations in activity were considerable among glands tested on the same day and among measurements from the same gland on different days. Farnesoic acid-stimulated rates of JH-III production were always higher than the basal rates, suggesting that the CA were not maximally activated. However, freshly excised low-activity CA, whose hormone production increased in the long-term conditions, showed similar farnesoic acid-stimulated rates of JH-III production to those of freshly excised high-activity glands, suggesting that at the time of excision of the corpora allata rate-limiting step(s) preceding farnesoic acid biosynthesis were inhibited or refractory to stimulation in vivo.     

Mating and diapause in hibernating adults ofMenida scotti puton (Heteroptera: Pentatomidae)

InMenida scotti aduts which mate during hibernation, we examined the effect of photoperiod on diapause induction in terms of the developmental degree of reproductive organs and corpora allata. In a hibernating population, mating season and physiological changes related to dipause were investigated. When newly emerged adults were reared under long- or short-day conditions, reservoir of the ectodermal accessory gland and corpora allata in males developed, while the developments of ovary and corpora allata in females were suppressed under both conditions. This suggests that diapause of this species is induced only in females independently photoperiodic conditions during adult stage. In a hibernating population, mating was observed from early to late November and from late March to early May. Observations of the development of reproductive organs and corpora allata suggest that diapause is induced only in females in the former period, while both sexes are not in diapause in the latter period.     

Reproductive biology of the German cockroach,Blattella germanica: Juvenile hormone as a pleiotropic master regulator

Juvenile hormone (JH) exerts major pleiotropic effects on cockroach development and reproduction. The production of JH by the corpora allata (CA) in the adult female German cockroach, Blattella germanica, is dependent upon and modulated by both internal and environmental stimuli. Mating, intake of high-quality food, social interactions, and the presence of vitellogenic ovaries facilitate JH synthesis. Conversely, starvation, deficient diets, enforced virginity, isolation, and a pre- or post-vitellogenic ovary cause the CA to produce less JH. Sensory stimulation of the genital vestibulum by the ootheca also inhibits the CA via signals that ascend the ventral nerve cord. All these stimulatory and inhibitory signals are integrated by the brain, and a preponderance of favorable signals results in a graded lifting of brain inhibition, permitting the synthesis and release of JH. The effects of inhibitory signals on JH biosynthesis can be lifted experimentally by severing nervous connections between the brain and the CA. Such an operation accelerates activation of the CA. Besides controlling gonadal maturation in females, JH concurrently regulates the production of sexual signals, including both attractant- and courtship-eliciting pheromones, and the behavioral expression of calling (pheromone release) and sexual receptivity. Although JH is required for the expression of copulatory readiness in female B. germanica, it appears that signals associated with copulation (spermatophore, sperm, accessory secretions) can inhibit this behavioral state even when titers of JH are permissive for receptivity. These observations suggest that JH might regulate sexual receptivity in females indirectly through other directives. In males, JH accelerates not only the onset of sexual readiness but also synthesis of accessory reproductive products. Lastly, we present a novel cockroach control strategy that is based on the intimate association between food intake and rising JH titers in B. germanica females. JH analogs cause abortion of fertile oothecae in gravid females. In turn, rising JH titers and vitellogenic oocytes induce feeding in females. With strategic placement of insecticidal baits and JH analogs, gravid females, which normally feed little and are difficult to control, can thus be effectively targeted for elimination. Arch. Insect Biochem. Physiol. 35:405–426, 1997. © 1997 Wiley-Liss, Inc.     

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.     

The allatostatic effect of 20-hydroxyecdysone on the adult viviparous cockroach, Diploptera punctata

The effect of 20-hydroxyecdysone upon the activity of corpora allata (CA) from female Diploptera punctata has been investigated. This ecdysteroid inhibits juvenile hormone (JH) biosynthesis by the CA, whether they have been implanted into a male, or remained in situ within the female. In the female, this inhibition is reflected in reduced oöcyte growth and vitellin content. The allatostatic effect of 20-hydroxyecdysone becomes apparent in vivo within 24 hr. However, no inhibition was observed when the CA were maintained in vitro for 42 hr in medium containing up to 1·10^?5 M 20-hydroxyecdysone. This suggests that the effect of the hormone upon the CA is indirect. These experiments raise the possibility that ecdysteroids play an allatostatic role during the normal gonotrophic cycle in Diploptera.     

保幼激素及其代谢产物的HPLC分离方法的改进和应用

传统的正相高效液相色谱（normal phase high performance liquid chromatography, NP-HPLC）可以较好地分离保幼激素（juvenile hormone, JH）Ⅰ、Ⅱ和Ⅲ,但不能分离保幼激素的代谢产物及其类似物。经过改进的反相高效液相色谱（reverse phase high performance liquid chromatography, RP-HPLC）不仅可以很好地分离保幼激素,还能定性定量分析保幼激素的代谢产物及其类似物。离体培养昆虫咽侧体（corpora allata, CA）所合成的、被同位素标记的痕量保幼激素可以用以上两种色谱方法进行分离和鉴定。此外,RP-HPLC还可以用来分离体内或体外同位素标记的保幼激素代谢产物,以及测量血淋巴中的保幼激素滴度。     

Vitellogenin induced in adult male Diploptera punctata by juvenile hormone and juvenile hormone analogue: Identification and quantitative aspects

The effect of the juvenile hormone analogue hydroprene on the appearance in the haemolymph of the yolk-protein precursor vitellogenin in male Diploptera punctata has been assessed using rocket immuno-electrophoresis. Vitellogenin was induced in a dose-dependent fashion, with a minimum dose of 10 μg hydroprene required for its appearance. Implantation of male corpora allata into male and female hosts also resulted in the appearance of vitellogenin in the haemolymph of the hosts, with females showing apparent greater sensitivity to the implantation.The identity of vitellogenin in male haemolymph was further confirmed using Ouchterlony double immunodiffusion and SDS polyacrylamide gel electrophoresis of immunoprecipitates. The electrophoretic pattern of immunoprecipitated haemolymph from females and hydroprene-treated males was similar, further confirming that the immunoprecipitable product was in fact vitellogenin.     

昆虫神经肽allatostati与allatotropin的研究新近展

Recently, two families of insect neuropeptides, the allatostatin, andallatotropin, have been identified. All allatostatins and allatotropins identified so farare neurosecretory polypeptides. A 13-amino acid allatotropin has been identifiedfrom adult Manduca sexta. A group of five structurally related allatostatins has beenidentified from Diploptara punctata. They either inhibit (allatostatin) or stimulate(allatotropin) the production of JH by CA. In this paper recent advances in researchon insect neuropeptides AS and AT and their biological significance are reported.     

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

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

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.     

昆虫神经肽allatostatin与allatotropin的研究新近展

昆虫神经肽ａｌｌａｔｏｓｔａｔｉｎ与ａｌｌａｔｏｔｒｏｐｉｎ的研究新近展关雪辰（中国科学院动物研究所北京１０００８０）昆虫神经激素是肽类激素,它控制昆虫许多关键的生理过程,例如生长、变态、生殖和行为等。保幼激素（ＪＨ）对昆虫的卵子发生成熟起着重要的调...