Hormonal modulation of reproduction and fertility signaling in polistine wasps

In social insects, it has been suggested that reproduction and the production of particular fertility-linked cuticular hydrocarbons (CHC) may be under shared juvenile hormone (JH) control, and this could have been key in predisposing such cues to later evolve into full-fledged queen pheromone signals. However, to date, only few studies have experimentally tested this “hormonal pleiotropy” hypothesis. Here, we formally test this hypothesis using data from four species of Polistine wasps, Polistes dominula, Polistes satan, Mischocyttarus metathoracicus, and Mischocyttarus cassununga, and experimental treatments with JH using the JH analogue methoprene and the anti-JH precocene. In line with reproduction being under JH control, our results show that across these four species, precocene significantly decreased ovary development when compared with both the acetone solvent-only control and the methoprene treatment. Consistent with the hormonal pleiotropy hypothesis, these effects on reproduction were further matched by subtle shifts in the CHC profiles, with univariate analyses showing that in P. dominula and P. satan the abundance of particular linear alkanes and mono-methylated alkanes were affected by ovary development and our hormonal treatments. The results indicate that in primitively eusocial wasps, and particularly in Polistes, reproduction and the production of some CHC cues are under joint JH control. We suggest that pleiotropic links between reproduction and the production of such hydrocarbon cues have been key enablers for the origin of true fertility and queen signals in more derived, advanced eusocial insects.     

Queen-signal modulation of worker pheromonal composition in honeybees

Worker sterility in honeybees is neither absolute nor irreversible. Whether under queen or worker control, it is likely to be mediated by pheromones. Queen-specific pheromones are not exclusive to queens; workers with activated ovaries also produce them. The association between ovarian activation and queen-like pheromone occurrence suggests the latter as providing a reliable signal of reproductive ability. In this study we investigated the effect of queen pheromones on ovary development and occurrence of queen-like esters in workers' Dufour's gland. Workers separated from the queenright compartment by a double mesh behaved like queenless workers, activating their ovaries and expressing a queen-like Dufour's gland secretion, confirming that the pheromones regulating both systems are non-volatile. Workers with developed ovaries produced significantly more secretion than sterile workers, which we attribute primarily to increased ester production. Workers separated from the queenright compartment by a single mesh displayed a delayed ovarian development, which we attribute to interrupted transfer of the non-volatile pheromone between compartments. We suggest that worker expression of queen-like characters reflects a queen-worker arms race; and that Dufour's gland secretion may provide a reliable signal for ovarian activation. The associative nature between ovary development and Dufour's gland ester production remains elusive.     

Reproductive plasticity in Polistes paper wasp workers and the evolutionary origins of sociality

Regulatory pathways in solitary species provide the raw materials for the evolution of sociality. Therefore, comparing the mechanisms that mediate reproductive plasticity in social species and their solitary ancestors can provide insight into the evolutionary origin of sociality. In many solitary insects, the effect of juvenile hormone (JH) on fertility is mediated through the fat body; individuals in good physical condition show a stronger fertility response to JH than individuals in poor physical condition. Here, we test whether a similar, condition-dependent JH response mediates fertility in workers of the primitively eusocial Polistes dominulus wasps. We test how body weight, JH, and adult nutrition influence worker ovarian development. Both JH-treatment and adult nutrition dramatically increased ovarian development. Body weight also influenced ovarian development, as large workers developed more eggs than smaller workers. Body weight and fat are strongly linked in P. dominulus workers, so these results suggest that the fat-dependent JH responsiveness common in solitary insects is conserved in social wasps. The simple, ancestral relationship between reproductive investment and physical condition may facilitate cooperation by allowing workers to adaptively allocate energy into reproduction based on their probability of successfully becoming a queen.     

Enhancing male sexual success in a lekking fly (Anastrepha suspensa Diptera: Tephritidae) through a juvenile hormone analog has no effect on adult mortality

While defending lek-territories, male Anastrepha suspensa (Loew) produce chemical, acoustic and visual courtship signals. In the laboratory and under semi-natural conditions, topical application of the juvenile hormone analog methoprene doubles pheromone production and subsequently doubles sexual success. However, sexual signals and interactions are likely to be physiologically expensive and so result in higher male mortality. Comparison of males kept in isolation for 35 days, but provided daily with a potential mate or a rival male, revealed that both male- and female-interactors shortened focal-male lifespan. In addition, focal males were either treated with methoprene or not, then either provided with protein in their sucrose-based diet or not. Protein proved to similarly double sexual success and also resulted in longer male life spans in all of the interactor-categories. However, there was no evidence that methoprene induced hypersexuality resulted in higher rates of mortality, i.e., the longevity of males treated with methoprene did not significantly differ from untreated males in the same interactor/diet categories. This apparent lack of costs to a putatively sexually selected signal is unexpected but presents an opportunity to increase the sexual competence of sterile flies with few consequences to their survival following mass-release.     

Soldier determination in Pheidole bicarinata: Effect of methoprene on caste and size within castes

Topical application of methoprene to final-instar larvae of the ant Pheidole bicarinata can induce soldier development. Soldier induction takes place if methoprene levels are above a soldier-determining threshold during a critical period of juvenile hormone-sensitivity that occurs during about days 4–6 of the final instar. Furthermore, the amount of exogenous methoprene applied affects the timing of metamorphosis and the adult size in both the minor worker and soldier castes. When larvae that receive methoprene treatment become minor workers these are always larger than acetone-treated controls. In larvae that become soldiers, growth and timing of metamorphosis vary with the dose of methoprene, but in a more complex way. A high dose of methoprene produces a metamorphic delay and large soldiers. However, the lowest effective dose for soldier induction produces early metamorphosis and small adults. On the basis of these results, we have expanded our model of a mechanism by which juvenile hormone could control determination of worker castes in Pheidole bicarinata.     

Ovarian activity correlates with extreme changes in cuticular hydrocarbon profile in the highly polygynous ant, Linepithema humile

In social insects, cuticular hydrocarbons of adults may vary with ovarian activity. Such variations are suggested to function in the regulation of reproduction within colonies. The modification of the CH profile with ovarian activity is usually interpreted as a signal of fertility causing workers to refrain from reproducing in the presence of the queen. We examined the effect of ovarian activity on the CH profiles in the Argentine ant Linepithema humile, a species where workers lack ovaries and are completely sterile. Our data show considerable differences in the CH profiles between fertile and infertile individuals. These differences are mainly qualitative. The CH profile strongly changes at the start of egg laying, both in mated and unmated queens. These results show that variation in cuticular hydrocarbons with ovarian activity in ants is not restricted to species with worker reproduction. We propose that in the Argentine ant, the cuticular hydrocarbons of laying queens correspond to a signal of fertility involved in the regulation of various aspects of reproduction, such as the rearing of new sexuals.     

Queen pheromones affecting the production of queen-like secretion in workers

The honeybee queen pheromones promote both worker sterility and worker-like pheromone composition; in their absence workers become fertile and express the queen pheromones. Which of the queen pheromones regulate worker pheromone expression and how, is still elusive. Here we investigated how two queen pheromones, the mandibular and Dufour’s, singly or combined, affect worker ovarian activation and occurrence of queen-like Dufour’s esters. Although queen mandibular pheromone (QMP) alone, or combined with Dufour’s secretion, inhibited to some extent worker reproduction, neither was as effective as the queen. The effect of the queen pheromones on worker pheromone expression was limited to workers with developed ovaries. Here too, QMP and Dufour’s combined had the greatest inhibitory effect. In contrast, treatment with Dufour’s alone resulted in augmentation of esters in the workers. This is another demonstration that a pheromone emitted by one individual affects the rates of its production in another individual. Ester production was tightly coupled to ovarian development. However fertile workers from queenright or QMP-treated colonies had significantly higher amounts of esters in their Dufour’s gland than untreated queenless colonies. The fact that the queen or QMP exert greater suppression on signal production than on ovary activation, suggests disparate regulatory pathways, and presents a challenging ultimate as well as proximate questions.     

Ecdysteroid fluctuations in adult Drosophila melanogaster caused by elimination of pupal reserves and synthesis by early vitellogenic ovarian follicles

Radioimmunoassay (RIA) of whole body extracts of Drosophila melanogaster males and females demonstrates that at eclosion all individuals contain high levels of ecdysteroid. Highly polar ecdysteroids (presumably metabolites) in the meconium represent approximately half of the total ecdysteroid RIA-activity present at this time and are subsequently eliminated. Ecdysteroids remaining after the elimination of the meconium are also highly polar as shown by reverse-phase high pressure liquid chromatography (RP-HPLC). The amount of ecdysteroid RIA-activity found in whole body extracts declines in both sexes until 18 h post-eclosion when levels begin to increase in the female and drop to undetectable levels in the male. In the female the ovaries are the major source of ecdysteroid. The increase in whole body ecdysteroid in the female coincides with the initiation of ovarian ecdysteroid production and accumulation. Topical application of methoprene, a juvenile hormone (JH) analog, stimulates ovarian ecdysteroid synthesis in apterous-ts 78j (ap^{ts 78j}), a temperature-sensitive juvenile hormone-deficient mutant, corroborating previous results suggesting a role of juvenile hormone in ovarian ecdysteroid production. Stage 8–9 follicles, whose development is juvenile hormone dependent, are shown to be the most active in ecdysteroid production. The regulatory potential of these stages is discussed.     

Interaction of pheromones during food exploitation by the termite Schedorhinotermes lamanianus

Abstract. Chemical signals from secretions of different exocrine glands modulate a variety of behavioural patterns in termite societies. These signals have multiple functions and may be interactive. During food exploitation workers of the African termite Schedorhinotermes lamanianus (Isoptera: Rhinotermitidae) employ, on foraging trails, the secretion from the sternal gland both for orientation and recruitment to a food source. The secretion from the labial gland, released onto the food by gnawing termites, stimulates additional workers to gnaw at the same site, thereby forming aggregations of gnawing termites. An interaction between these two pheromones during food exploitation is demonstrated for the first time. The volatile signal from the sternal gland inhibits in a dose-dependent manner the non-volatile, highly persistent, signal from the labial gland. The development of gnawing aggregations is inhibited and established ones are dissolved. Behavioural evidence for the perception of both the volatile signal from the sternal gland by olfactory neurones and of the non-volatile signal from the labial gland by gustatory neurones on the antennae is given. The interaction of the two pheromones as a basis for the development of distinct commuting and gnawing zones on the food source, and as a means for a dynamic regulation of food exploitation, is discussed.     

Sex, age and ovarian activity affect cuticular hydrocarbons in Diacamma ceylonense, a queenless ant

In the queenless ant, Diacamma ceylonense, the cuticular hydrocarbons (C25-C35) of nestmate workers vary in their proportions according to age and fertility. Newly eclosed adults ('callows') initially have the same cuticular profile, but with time this changes to that typical of foragers. In contrast, workers that begin to produce eggs develop a different cuticular profile. Several substances (n-C29 and some methyl C25 and C27) discriminate these different social categories (callows, foragers and egg-layers). In Diacamma ceylonense, inter-colony variation of the cuticular hydrocarbons was much lower than intra-colony variation. We also found qualitative differences between the sexes, with males having a clearly different profile with much more alkanes. We discuss these results in the context of physiological models of the relation between ovarian activity and the synthesis of cuticular hydrocarbons. Variations in cuticular profile are a reliable reflection of ovarian activity, and could be used by ants as a fertility signal.     

Methoprene does not affect juvenile hormone titers in honey bee (Apis mellifera) workers

Methoprene, a juvenile hormone (JH) analog, is a widely used insecticide that also accelerates behavioral development in honey bees (Apis mellifera). JH regulates the transition from nursing to foraging in adult worker bees, and treatment with JH or methoprene have both been shown to induce precocious foraging. To determine how methoprene changes honey bee behavior, we compared JH titers of methoprene‐treated and untreated bees. Behavioral observations confirmed that methoprene treatment significantly increased the number of precocious foragers in 3 out of 4 colonies. In only 1 out of 4 colonies, however, was there a significant difference in JH titers between the methoprene‐treated and control bees. Further, in all 4 colonies, there was no significant differences in JH titers between precocious and normal‐aged foragers. These results suggest that methoprene did not directly affect the endogenous JH secreted by corpora allata. Because methoprene caused early foraging without changing workers’ JH titers, we conclude that methoprene most likely acts directly on the JH receptors as a substitute for JH.     

Reproductive effects in German cockroaches by ecdysteroid agonist RH-0345, juvenile hormone analogue methoprene and carbamate benfuracarb

Blatta germanica is the more prevalent cockroach species in Algeria. In the present study, we tested the effect on reproduction in B. germanica of two insect growth regulators, RH-0345, a benzoylhydrazine analogue that mimics the action of 20-hydroxyecdysone, and methoprene, one of the most commercially important juvenile hormone analogues, and a novel carbamate insecticide, benfuracarb. The compounds were applied topically (10 and 20 microg/insect for RH-0345, and 1 and 10 microg/insect for methoprene) or orally administrated (at 2% for benfuracarb) on newly emerged females and evaluated on reproductive events during the adult life (2, 4 and 6 days). Treatment with RH-0345 and benfuracarb reduced significantly the number of oocytes, the size and the volume of the basal oocyte during the experimental period. Methoprene distorted the ovarian development since it caused a significant reduction in the number of oocytes at 2, 4 and 6 days for the two tested doses, and an increase in oocyte size at 2, 4 and 6 days with 1 microg and a decrease with 10 microg. In a second series of experiments, the effects of these compounds were assayed on the ovarian proteins. Data from biochemical analysis revealed that RH-0345 and benfuracarb reduced the ovarian amounts of proteins, while treatment with methoprene increased it during the sexual maturation.     

Effects of precocene and methoprene application in young adult Musca autumnalis

Abstract Observations were made on the relative rates of growth among corpora allata, ovarian follicles and fat bodies in diapausing and reproductive Musca autumnalis DeGeer. Rates of corpora allata growth were greater in long-day than in short-day animals. Topically applied precocene II reduced the size of the corpora allata. Two sequential doses of precocene inhibited vitellogenesis. Topically applied methoprene produced vitellogenesis in short-day flies and inhibited development of hypertrophied fat body. This work provides evidence that Juvenile Hormone is the primary agent controlling diapause and non-diapause (vitellogenic) development in the ovaries of face flies.     

天蚕卵黄发生的激素调控

报告了蜕皮激素和保幼激素对天蚕Antheraea yamamai卵黄发生的调控作用。当单独以20－羟基蜕皮酮或保幼激素类似物methoprene处理,以及同时用这两种激素处理天蚕蛹时,蛹期脂肪体和血淋巴中卵黄原蛋白（Vg)含量明显高于对照,即二对Vg的合成起促进作用。然而,卵巢中卵黄蛋白（Vt)含量则因激素种类而异,以保幼激素处理时明显低于对照,以20－羟基蜕皮酮处理则反之,即前抑制卵巢对Vg的摄取,而后则起促进作用。离体培养脂肪体并以激素处理的结果表明,20－羟基蜕皮酮和methoprene均能促进Vg合成,但前作用更。综合考虑上述结果可以认为蜕皮激素对该蚕的卵黄发生起主要调控作用。     

QUEEN SIGNALING IN SOCIAL WASPS

Social Hymenoptera are characterized by a reproductive division of labor, whereby queens perform most of the reproduction and workers help to raise her offspring. A long‐lasting debate is whether queens maintain this reproductive dominance by manipulating their daughter workers into remaining sterile (queen control), or if instead queens honestly signal their fertility and workers reproduce according to their own evolutionary incentives (queen signaling). Here, we test these competing hypotheses using data from Vespine wasps. We show that in natural colonies of the Saxon wasp, Dolichovespula saxonica, queens emit reliable chemical cues of their true fertility and that these putative queen signals decrease as the colony develops and worker reproduction increases. Moreover, these putative pheromones of D. saxonica show significant conservation with those of Vespula vulgaris and other Vespinae, thereby arguing against fast evolution of signals as a result of a queen–worker arms race ensuing from queen control. Lastly, levels of worker reproduction in these species correspond well with their average colony kin structures, as predicted by the queen signaling hypothesis but not the queen control hypothesis. Altogether, this correlative yet comprehensive analysis provides compelling evidence that honest signaling explains levels of reproductive division of labor in social wasps.     

Age and Behavior of Honey Bees,Apis mellifera (Hymenoptera: Apidae), that Perform Vibration Signals on Workers

The vibration signal is one of the most commonly occurring communication displays in honey bee (Apis mellifera) colonies. It may function in a ‘modulatory’ manner, because it causes a nonspecific increase in activity that enhances a variety of behaviors depending upon the age and caste of the recipient. We examined honey bee workers that performed vibration signals on other workers in three observation hives, each containing a population of marked bees of known age. In all three colonies, the mean age of the first performance of the vibration signal was significantly different from the mean age at which workers first performed waggle dances, carried pollen loads, or attended the queen. However, workers of all ages, except those less than 3 d old, could perform vibration signals. In older workers of foraging age, signal performance was most closely associated with recent foraging success. Younger workers that vibrated did not appear to be early-maturing foragers and thus their signals were probably not influenced by food collection. Rather, for these preforaging-age workers, signal performance was associated more with periods of orientation flight, during which younger bees learn the location of the nest and surrounding landmarks. Thus, the vibration signal may be triggered by different stimuli in different worker age classes. Because it elicits a general increase in activity in all recipients, the signal may help adjust many different colony behaviors simultancously to changes in foraging success and colony development.     

Queen pheromones in Temnothorax ants: control or honest signal?

Background  

The division of reproductive labor among group members in insect societies is regulated by "queen pheromones". However, it remains controversial whether these are manipulative, i.e., actively suppress worker reproduction, or honestly signal the fertility status of the queen to which workers react in their own interest by refraining from laying eggs. Manipulative queen control is thought to lead to an evolutionary arms race between queens and workers, resulting in complex queen bouquets that diverge strongly among different populations and species. In contrast, honest signals would evolve more slowly and might therefore differ less strongly within and among species.     

HORMONAL CONTROL OF THE VITELLOGENESIS IN THE JAPANESE OAK SILKWORM, ANTHERAEA YAMAMAZ (LEPIDOPTERA: SATURNIIDAE)

Abstract Effects of ecdysteroid and juvenile hormone (JH) on vitellogenesis of the Japanese oak silkworm, Antheraea yamami are reported in this article. After topical treatment with 20-hydroxyecdysone alone or JH analog (i.e. methoprene) alone and combined treatment with these two chemicals, vitellogenin (Vg) titers in the fat body and haemolymph at the pupal stage were mostly higher than those of the control, indicating that both ecdysteroid and JH exerted a promoting effect on the synthesis of Vg. In contrast, the Vg uptake was markedly inhibited by JH while stimulating effect of the ecdysteroid could be shown that vitellin (Vt) titer in the ovary was lower after methoprene treatments, but higher after 20-hydroxyecdyson treatments. Meanwhile, effects of these two hormones on Vg synthesis in the fat body were also tested with the incubation in vitro with Grace medium containing H-leucine and the hormones. The results demonstrated that Vg synthesis was stimulated after treating with methoprene alone or 20-hydroxyecdysone alone and combined treating with these two chemicals, and particularly ecdysteroid had more marked positive effect. To comprehensively concluded our results, it could be regarded that ecdysteroid play the main role in the regulation of vitellogenesis for the Japanese oak silkworm.