Imaginal Molt Research Articles

Dual function of the transcription factor Ftz-f1 on oviposition in the cockroach Blattella germanica.

The transcription factor Ftz-f1 has multiple functions in insect development in a spatial-temporal line. One of these roles is in the insect ovaries, specifically in the regulation of steroidogenic enzymes production. We studied the function of F in Blattella germanica oogenesis, as it shows two moments of high expression in ovaries: before the imaginal moult, and just before ovulation in the adult. Injecting dsftz-f1 into adult females, either just after the imaginal moult or just prior to choriogenesis, prevented oviposition, with differences between the two approaches. In 3-day-old adult females treated with dsftz-f1 just after the emergence, the expression of ftz-f1 was not modified, but the steroidogenic genes increased their expression. ftz-f1 transcript levels in the ovaries of 5-day-old dsftz-f1-treated females were significantly depleted, and the expression levels of the same steroidogenic genes began to decrease. These results suggest that Ftz-f1 regulates the expression of steroidogenic genes in B. germanica, with phm possibly being a key target. Ftz-f1 has a different temporal function in the cytoskeleton of follicular cells of the basal ovarian follicles. Early in the gonadotrophic cycle, Ftz-f1 promotes the expression of genes related to the cytoskeleton and muscle proteins, while at the end of the cycle it maintains the expression levels of these genes, thus ensuring correct ovulation.

Myofilaments promote wing expansion and maintain genitalia morphology in the American cockroach, Periplaneta americana.

Insects are the most widely distributed and successful animals on the planet. A large number of insects are capable of flight with functional wings. Wing expansion is an important process for insects to achieve functional wings after eclosion and healthy genital morphology is crucial for adult reproduction. Myofilaments are functional units that constitute sarcomeres and trigger muscle contraction. Here, we identified four myofilament proteins, including Myosin, Paramyosin, Tropomyosin and Troponin T, from the wing pads of nymphs in the American cockroach, Periplaneta americana. RNAi-mediated knockdown of Myosin, Paramyosin, Tropomyosin and Troponin T in the early stage of final instar nymphs caused a severely curly wing phenotype in the imaginal moult, especially in the Paramyosin and Troponin T knockdown groups, indicating that these myofilament proteins are involved in controlling wing expansion behaviours during the nymph-adult transition. In addition, the knockdown resulted in abnormal external genitalia, caused ovulation failure, and affected male accessory gland development. Interestingly, the expression of myofilament genes was induced by methoprene, a juvenile hormone (JH) analogue, and decreased by the depletion of the JH receptor gene Met. Altogether, we have determined that myofilament genes play an important role in promoting wing expansion and maintaining adult genitalia morphology, and their expression is induced by JH signalling. Our data reveal a novel mechanism by which wing expansion is regulated by myofilaments and the functions of myofilaments are involved in maintaining genitalia morphology.

Preliminary larvicidal effect of seed oil from Cleome arabica L. on fifth instar larvae of Schistocerca gregaria (Orthoptera: Acrididae)

In this present study, the toxicity of Cleome arabica L. (Capparidaceae) seed oils was tested on L5 larvae of Schistocerca gregaria, Forsk. (Orthoptera-Acrididae). The forced administration of 60 ?L/individual of this vegetable oil leads to the death of all the treated larvae either before or after a few days of exuviation (imaginal moult). Signs of intoxication were observed a few hours after treatment, i.e. movement disorders, diarrhoea, inability to take food and weight loss. Male L5 larvae seem more sensitive than female L5; the mortality rates being around 90.91% and 36.36% respectively after 16 days. The TL50 recorded in treated L5 larvae is around 11 days.

Hormone-related genes heterochronically and modularly regulate neotenic differentiation in termites

Caste development in social insects requires the coordination of molting and metamorphosis during postembryonic development. In termites, i.e., hemimetabolous eusocial insects, caste fate is determined during postembryonic development. However, it is not fully understood how the mechanisms of molting/metamorphosis are regulated in the course of differentiation between reproductive and sterile castes. In termites, only reproductives derived from alates are imagos and other sterile castes (including developmentally-terminal soldier caste) are basically juveniles or nymphs. Furthermore, supplementary reproductives that appear when the original queens and kings die or become senescent, exhibit larval features such as winglessness, and are called neotenics. Therefore, the question of whether neotenics are larvae or imagos is still under debate. In this study, by inducing female neotenic differentiation in a damp-wood termite Hodotermopsis sjostedti, morphological investigations together with juvenile hormone (JH) quantification and expression/functional analyses of genes responsible for molting and/or metamorphosis were carried out. JH titer and expression of one of the downstream genes (Kr-h1) were shown to be temporarily lowered, but increased just prior to the molt into neotenics, while consistently lowered in imaginal molt (i.e., alate differentiation). In contrast, ecdysone-related genes (EcR and E93) were upregulated at both neotenic and alate differentiation, suggesting that the heterochronic actions of ecdysone and JH lead the neotenic differentiation. Moreover, expression analyses, supported by reverse genetic experiments, showed that EcR and E93 were specifically upregulated in genital sternites (EcR and E93) and ovaries (E93) and required for the development of imaginal characters. These results suggest that the resultant mosaic phenotype of female neotenics is due to modular responses of different body parts to hormonal actions.

LARVICIDAL EFFECT OF THE SEED OILS OF TWO INDIGENOUS PLANTS FROM THE ALGERIAN SAHARA ON THE DESERT LOCUST

The lethal and sublethal effects of the seed oils of two indigenous plants of the Algerian Sahara, Peganum harmala L. (Zygophyllaceae) and Datura stramonium L. (Solanaceae), were investigated. Administration of 60 μl of oil by forced oral injection using a micropipette to the L5 larvae of Schistocerca gregaria Forsk. (Orthoptera: Acrididae) revealed the deterrent effect of these oils on treated larvae. The treatment resulted in various toxicological symptoms, such as intense defecation, diarrhea, weight loss, reduction in motor activity, delay and difficulty in molting and, in the most extreme cases, the death of treated individuals. During the treatment of L5 larvae of S. gregaria with P. harmala seed oil, various toxicological symptoms appeared: 81.81% of individuals presented with diarrhea; 68.18% of individuals lost weight; 72.72% exhibited reduced motor activity, and 100% of surviving individuals experienced a delay in their molt. On the other hand, in L5 larvae treated with D. stramonium seed oil, 77.27% of individuals had diarrhea, 100% showed weight loss and 100% of individuals reduced their motor activity. D. stramonium seed oil has been shown to be more toxic than P. harmala seed oil. The oral administration of 60 μl of D. stramonium seed oil caused the blocking of the phenomenon of exuviation in 100% of the treated L5 larvae, resulting in death after 16 days. While P. harmala seed oils caused 50% mortality after 12 days, the 50% surviving individuals were able to complete their imaginal molt with difficulties, which resulted in malformations. The estimated lethal time 50 (LT50) in larvae (L5) treated with D. stramonium seed oil was 3.67 days. It was more toxic than the LT50 obtained in larvae (L5) treated with the oil of P. harmala seeds, which was 12 days. The food intake in L5 larvae of S. gregaria treated with D. stramonium seed oil was 0.28 ± 0.18 g/day, it was lower than the average daily consumption recorded in the L5 larvae treated with P. harmala seed oil, which was 0.67 ± 0.36 g/day, D. stramonium seed oil appears to be more toxic, and profoundly affects food intake. The values of the apparent digestive utilization coefficient (DUC a) reported for L5 larvae treated with seed oil of P. harmala and D. stramonium were 39.32 ± 13.07% and 34.23 ± 29,07%, respectively. These values were significantly lower compared to the control group value, which was 70.63 ± 19.56%. Likewise, the digestive conversion coefficients (CCD) recorded in the L5 of S. gregaria treated with the seed oils of P. harmala and D. stramonium were -75.07 ± 54.45% and -3.08 ± 1.18, respectively. However, in the control group of L5 larvae, the noted CCD was 1.004 ± 0.073. Values of the consumption index (CI) reported for L5 larvae treated with the seed oils of P. harmala and D. stramonium were low, 6.74 ± 4.45 and 3.82 ± 2.45, respectively, while for the L5 larvae of the control group, it was 15.74 ± 3.51.

Previous flight facilitates partner finding in female crickets

In the cricket Gryllus bimaculatus, flying occurs soon after the last imaginal molt and precedes the mating behavior in natural conditions. Here, we tested the hypothesis that flying may improve subsequent behavioral performance in a novel environment in female crickets. We developed a behavioral set-up to test female cricket responsiveness to male calling song as well as their ability to locate and find the source of the song. The male song was produced by a loudspeaker hidden behind the fabric wall of a spacious square arena. Forced flight prior to the test promoted female sexual searching behavior in the novel environment. After the flight, more females reached the hidden source zone, spent more time near the source and finally more of them climbed over the wall section immediately in front of the hidden loudspeaker. At the same time, their behavior in the arena did not differ from the control group when the calling song was not delivered, suggesting that flight exerts its behavioral effects by influencing sexual motivation. Our results support the suggestion that preceding intense locomotion facilitates sexual searching behavior of females in a novel environment.

Structure and development of the complex helmet of treehoppers (Insecta: Hemiptera: Membracidae)

Some insects possess complex three-dimensional (3D) structures that develop under the old cuticle prior to the last imaginal molt. Adult treehoppers (Insecta: Hemiptera: Auchenorrhyncha: Membracidae) have one such complex 3D structure, known as a helmet, on their dorsal side. The adult helmet likely forms inside the nymphal pronotum during the final instar nymphal stage.Previous morphological studies have reported that the adult helmet is a large, bi-layered, plywood-like structure, whereas the nymphal pronotum is a monolayer, sheath-like structure. The adult helmet is much larger than nymphal helmet. Thus, the emergence of the adult helmet involves two structural transitions: a transition from a monolayer, sheath-like pronotum to a bi-layer, plywood-like helmet, and a transition in size from small to large. However, when, how, and in what order these transitions occur within the nymphal cuticle is largely unknown.To determine how adult helmet development occurs under the nymphal cuticle, in the present study we describe the morphology of the final adult helmet and investigate developmental trajectories of the helmet during the final instar nymphal stage. We used micro-CT, scanning electron microscope and paraffin sections for morphological observations, and used Antianthe expansa as a model species.We found that the structural transition (from monolayer, sheath-like structure to bi-layer, roof-like structure) occurs through the formation of a “miniature” of the adult helmet during the middle stage of development and that subsequently, extensive folding and furrows form, which account for the increase in size. We suggest that the making of a “miniature” is the key developmental step for the formation of various 3D structures of treehopper helmets.

Aimed limb movements in a hemimetabolous insect are intrinsically compensated for allometric wing growth by developmental mechanisms.

ABSTRACTFor aimed limb movements to remain functional, they must be adapted to developmental changes in body morphology and sensory-motor systems. Insects use their limbs to groom the body surface or to dislodge external stimuli, but they face the particular problem of adapting these movements to step-like changes in body morphology during metamorphosis or moulting. Locusts are hemimetabolous insects in which the imaginal moult to adulthood results in a sudden and dramatic allometric growth of the wings relative to the body and the legs. We show that, despite this, hind limb scratches aimed at mechanosensory stimuli on the wings remain targeted to appropriate locations after moulting. In juveniles, the tips of the wings extend less than halfway along the abdomen, but in adults they extend well beyond the posterior end. Kinematic analyses were used to examine the scratching responses of juveniles (fifth instars) and adults to touch of anterior (wing base) and posterior (distal abdomen) targets that develop isometrically, and to wing tip targets that are anterior in juveniles but posterior in adults. Juveniles reach the (anterior) wing tip with the distal tibia of the hind leg using anterior rotation of the thoraco-coxal and coxo-trochanteral (‘hip’) joints and flexion of the femoro-tibial (‘knee’) joint. Adults, however, reach the corresponding (but now posterior) wing tip using posterior rotation of the hip and extension of the knee, reflecting a different underlying motor pattern. This change in kinematics occurs immediately after the adult moult without learning, indicating that the switch is developmentally programmed.

Factors affecting the egg-laying pattern of Forficula auricularia (Dermaptera: Forficulidae) in three climatologically different zones of North America

AbstractEgg-laying patterns of the European earwig, Forficula auricularia Linnaeus (Dermaptera: Forficulidae), from three climatologically different areas; Montréal, Québec, Canada, Vancouver, British Columbia, Canada, and San Francisco, California, United States of America; were compared in laboratory. Three different egg-laying patterns were observed. Among the biotic parameters studied; previtellogenic follicular atresia, number of oocytes per ovariole at imaginal moult, female life expectancy, and ovarian activity explained female successful responses to local weather conditions. Follicular atresia, combined with a low number of oocytes, impeded oviposition in a low percentage of females from Montréal. It also impedes the production of a third brood in the Vancouver population, and of a fourth brood in the San Francisco population. Both female life expectancy and length of ovarian activity also had a significant impact on the oviposition pattern in the three climatic regions. Under the cold temperate climate of the Montréal area, only one brood was produced due to the short period of female fertility. Longer period of fertility and life expectancy enabled females to produce two broods in the milder temperate climate of Vancouver, and three broods in the San Francisco climate. Our results strongly suggest an exaptation enabling the native European populations of F. auricularia to adapt successfully in North America.

Post-molting development of wind-elicited escape behavior in the cricket

Arthropods including insects grow through several developmental stages by molting. The abrupt changes in their body size and morphology accompanying the molting are responsible for the developmental changes in behavior. While in holometabolous insects, larval behaviors are transformed into adult-specific behaviors with drastic changes in nervous system during the pupal stage, hemimetabolous insects preserve most innate behaviors whole life long, which allow us to trace the maturation process of preserved behaviors after the changes in body. Wind-elicited escape behavior is one of these behaviors and mediated by cercal system, which is a mechanosensory organ equipped by all stages of nymph in orthopteran insects like crickets. However, the maturation process of the escape behavior after the molt is unclear. In this study, we examined time-series of changes in the wind-elicited escape behavior just after the imaginal molt in the cricket. The locomotor activities are developed over the elapsed time, and matured 24h after the molt. In contrast, a stimulus-angle dependency of moving direction was unchanged over time, meaning that the cercal sensory system detecting airflow direction was workable immediately after the molt, independent from the behavioral maturation. The post-molting development of the wind-elicited behavior was considered to result not simply from maturation of the exoskeleton or musculature because the escape response to heat-shock stimulus did not change after the molt. No effect of a temporal immobilization after the imaginal molt on the maturation of the wind-elicited behavior also implies that the maturation may be innately programmed without experience of locomotion.

Group-housed females promote production of asexual ootheca in American cockroaches

BackgroundFacultative parthenogenesis, seen in many animal phyla, is a reproductive strategy in which females are able to generate offspring when mating partners are unavailable. In some subsocial and eusocial insects, parthenogenesis is often more prevalent than sexual reproduction. However, little is known about how social cooperation is linked to the promotion of parthenogenesis. The domiciliary cockroach Periplaneta americana is well-suited to addressing this issue as this species belongs to the superfamily Blattoidea, which diverged into eusocial termites and shows facultative parthenogenesis.ResultsWe studied environmental factors that influence asexual production of ootheca using behavioral assays in P. americana. When more than three virgin females immediately after the imaginal molt were kept together in a small sealed container, they tended to produce egg cases (oothecae) via parthenogenesis earlier than did isolated females, resulting in apparent synchronization of ootheca production, even among females housed in different containers. In contrast, virgin females housed with genitalia-ablated males or group-housed females with antennae ablated did not significantly promote ootheca production compared to isolated females. Daily addition of the primary sex pheromone component to the container did not promote ootheca production in isolated females. Another line of study showed that grouped females make parthenogenesis more sustainable than previously known; a founder colony of 15 virgin females was sufficient to produce female progeny for a period of more than three years.ConclusionsGroup-housed females promote and stabilize asexual ootheca production compared to isolated females, and that this promotion is triggered by female-specific chemosensory signals (other than sex pheromone) primarily detected by antennae. Promotion of ootheca production between females is likely to be an early stage of social cooperation, reminiscent of the foundation and maintenance of a colony by female pairs in the eusocial termite Reticulitermes speratus.

Juvenile hormone biosynthesis in adult Blattella germanica requires nuclear receptors Seven-up and FTZ-F1

In insects, the transition from juvenile development to the adult stage is controlled by juvenile hormone (JH) synthesized from the corpora allata (CA) glands. Whereas a JH-free period during the last juvenile instar triggers metamorphosis and the end of the growth period, the reappearance of this hormone after the imaginal molt marks the onset of reproductive adulthood. Despite the importance of such transition, the regulatory mechanism that controls it remains mostly unknown. Here, using the hemimetabolous insect Blattella germanica, we show that nuclear hormone receptors Seven-up-B (BgSvp-B) and Fushi tarazu-factor 1 (BgFTZ-F1) have essential roles in the tissue- and stage-specific activation of adult CA JH-biosynthetic activity. Both factors are highly expressed in adult CA cells. Moreover, RNAi-knockdown of either BgSvp-B or BgFTZ-F1 results in adult animals with a complete block in two critical JH-dependent reproductive processes, vitellogenesis and oogenesis. We show that this reproductive blockage is the result of a dramatic impairment of JH biosynthesis, due to the CA-specific reduction in the expression of two key JH biosynthetic enzymes, 3-hydroxy-3-methylglutaryl coenzyme A synthase-1 (BgHMG-S1) and HMG-reductase (BgHMG-R). Our findings provide insights into the regulatory mechanisms underlying the specific changes in the CA gland necessary for the proper transition to adulthood.

The molecular sensory machinery of a Chagas disease vector: expression changes through imaginal moult and sexually dimorphic features

The triatomine bug Rhodnius prolixus is a main vector of Chagas disease, which affects several million people, mostly in Latin-America. Host searching, pheromone communication, and microclimatic preferences are aspects of its behaviour that depend on multimodal sensory inputs. The molecular bases of these sensory processes are largely unknown. The expression levels of genes transcribed in antennae were compared between 5th instar larvae, and female and male adults by means of RNA-Seq. The antennae of R. prolixus showed increased expression of several chemosensory-related genes in imaginal bugs, while both sexes had similar expression patterns for most target genes. Few cases suggest involvement of target genes in sexually dimorphic functions. Most odorant and ionotropic receptor genes seemed to be expressed in all libraries. OBPs and CSPs showed very high expression levels. Other sensory-related genes such as TRPs, PPKs and mechanoreceptors had consistent levels of expression in all libraries. Our study characterises most of the sensory gene repertoire of these insects, opening an avenue for functional genetics studies. The increase in expression of chemosensory genes suggests an enhanced role in adult bugs. This knowledge allows developing new behaviour interfering strategies, increasing the options for translational research in the vector control field.

Smads and insect hemimetabolan metamorphosis

In contrast with Drosophila melanogaster, practically nothing is known about the involvement of the TGF-β signaling pathway in the metamorphosis of hemimetabolan insects. To partially fill this gap, we have studied the role of Smad factors in the metamorphosis of the German cockroach, Blattella germanica. In D. melanogaster, Mad is the canonical R-Smad of the BMP branch of the TGF-β signaling pathway, Smox is the canonical R-Smad of the TGF-β/Activin branch and Medea participates in both branches. In insects, metamorphosis is regulated by the MEKRE93 pathway, which starts with juvenile hormone (JH), whose signal is transduced by Methoprene-tolerant (Met), which stimulates the expression of Krüppel homolog 1 (Kr-h1) that acts to repress E93, the metamorphosis trigger. In B. germanica, metamorphosis is determined at the beginning of the sixth (final) nymphal instar (N6), when JH production ceases, the expression of Kr-h1 declines, and the transcription of E93 begins to increase. The RNAi of Mad, Smox and Medea in N6 of B. germanica reveals that the BMP branch of the TGF-β signaling pathway regulates adult ecdysis and wing extension, mainly through regulating the expression of bursicon, whereas the TGF-β/Activin branch contributes to increasing E93 and decreasing Kr-h1 at the beginning of N6, crucial for triggering adult morphogenesis, as well as to regulating the imaginal molt timing.

Reproductive Biology of Triatoma brasiliensis (Hemiptera, Triatominae) During the Imaginal Molt.

The triatomines are vectors of the protozoan Trypanosoma cruzi, etiologic agent of Chagas disease. These insects are sexually active after the imaginal molt. Some aspects have been studied in Triatoma brasiliensis during the imaginal molt, such as autogeny in virgin females and the relationship between blood ingestion by fifth instar nymph and the realization of the imaginal molt. Thus, to aid in the understanding of reproductive biology and developmental physiology of these vectors, this article analyzes the spermatogenesis of T. brasiliensis during the imaginal molt. The analysis of the seminiferous tubules from males in the fifth instar during imaginal molt has demonstrated that T. brasiliensis has only a few spermatids and a plentiful quantity of sperm. Thus, we suggest that during imaginal molt the cell division is disrupted aiming to reduce energy costs and the differentiation into sperm is stimulated to ensure the paternity of the adult male.

Caffeine effects on AdoR mRNA expression in Drosophila melanogaster

AbstractIn this study, we aimed to evaluate whether exposure to caffeine in the early stages of development affect AdoR mRNA expression levels in the fruit fly (Drosophila melanogaster) and how this will relate to the developmental success of flies. Adenosine receptors are seen as the most important biochemical targets of caffeine. Simultaneously adenosine signaling orchestrates the development and growth of insects. We demonstrate that AdoR mRNA expression in D. melanogaster is persistent from early stages till imago. Strong alterations in AdoR expression were observed in larvae that had been treated with caffeine. However, after the imaginal molt, the differences in AdoR expression between the insects from all of the test groups evened out despite a wide range of developmental success in the groups. Taken together, these results suggest that caffeine affects the expression of its cellular targets even from the early stages of fruit fly development and thus there is a significantly lower larvae-to-adult survival rate. Moreover, we also proved that the expression of AdoR undergoes a peculiar reset during the maturation of D. melanogaster despite the conditions in which larvae developed.

Molecular basis of peripheral olfactory plasticity in Rhodnius prolixus, a Chagas disease vector

Olfaction is fundamental for most animals and critical for different aspects of triatomine biology, including host-seeking, reproduction, avoidance of predators, and aggregation in shelters. Ethological and physiological aspects of these olfactory-mediated behaviors are well understood, but their molecular bases are still largely unknown. Here we investigated changes in molecular mechanisms at the peripheral olfactory level in response to different physiological and developmental conditions. For this, the antennal expression levels of the odorant (Orco) and ionotropic (IR8a, IR25a and IR76b) coreceptor genes were determined in Rhodnius prolixus by means of quantitative real-time PCR (qRT-PCR) analysis. Gene expression changes were analyzed to test the effect of feeding and imaginal molt for both sexes. Moreover, we analyzed whether expression of these genes changed during the early life of adult bugs. Under these conditions bugs display distinct behavioral responses to diverse chemical stimuli. A significantly decreased expression was induced by blood feeding on all coreceptor genes. The expression of all genes was significantly increased following the imaginal molt. These results show that olfactory coreceptor genes have their expression altered as a response to physiological or developmental changes. Our study suggests that olfactory coreceptor genes confer adaptability to the peripheral olfactory function, probably underlying the known plasticity of triatomine olfactory-mediated behavior.

Role of Methoprene-tolerant (Met) in adult morphogenesis and in adult ecdysis of Blattella germanica.

Juvenile Hormone (JH) represses metamorphosis of young instars in insects. One of the main players in hormonal signalling is Methoprene-tolerant (Met), which plays the role of JH receptor. Using the Polyneopteran insect Blattella germanica as the model and RNAi for transcript depletion, we have confirmed that Met transduces the antimetamorphic signal of JH in young nymphs and plays a role in the last nymphal instar moult in this species. Previously, the function of Met as the JH receptor had been demonstrated in the Eumetabola clade, with experiments in Holometabola (in the beetle Tribolium castaneum) and in their sister group Paraneoptera (in the bug Pyrrhocoris apterus). Our result shows that the function of Met as JH receptor is also conserved in the more basal Polyneoptera. The function of Met as JH transducer might thus predate the evolutionary innovation of metamorphosis. Moreover, expression of Met was also found in last nymphal instar of B. germanica, when JH is absent. Depletion of Met in this stage provoked deficiencies in wing growth and ecdysis problems in the imaginal moult. Down-regulation of the ecdysone-inducible gene E75A and Insulin-Like-Peptide 1 in these Met-depleted specimens suggest that Met is involved in the ecdysone and insulin signalling pathways in last nymphal instar, when JH is virtually absent.

Fertilization and fertility in the female of Varroa destructor, a key point for the parasite population dynamics

In Varroa destructor, the spermathecal content allows the female mite to carry out several successive reproductive cycles after a single mating event. The present study aimed to identify the main factors influencing the sperm content. Mite adult females were sampled at different bee colony phenological phases and on different host stages/sexes. Spermatozoa was characterized and counted. Spermatozoa maturity was acquired sufficiently fast (less than 5 days after the imaginal moult), their viability (ca. 91.5 %) and the fertilization success (ca. 1 spermatozoa spent per oocyte) were sufficiently high so that none of these parameters were limiting factors in the dynamics of mite populations. Moreover, no modulation was detected in the spermathecal content according to the brood status and the seasonal activity of the colony.

Post-Embryonic Development of Circadian Oscillations Within and Outside the Optic Lobe in the Cricket,Gryllus bimaculatus

The adult cricket Gryllus bimaculatus has a central clock in the optic lobe that regulates overt activity rhythms and secondary oscillators in the tissue outside the optic lobe. Here we investigated properties of the rhythmic expression of clock genes in the optic lobe and extra-optic lobe tissues in nymphs, and compared them with those of adults. In the optic lobe, mRNA of the clock genes period (per), timeless (tim), cycle (cyc) and Clock (Clk) were expressed in patterns similar to those in adult profiles, but at significantly lower levels. Among the extra-optic lobe tissues, the brain and TAG showed a rhythmic expression of per and tim, the mid-gut only in tim, and the anterior-stomach in none of the genes studied. The mRNA levels of clock genes were again significantly lower than those in adults. Unlike in adults, the brain and mid-gut lost their rhythms of clock gene expression in DD, and when the optic lobes were bilaterally removed. These results suggest that the rhythms outside the optic lobe are weak in nymphs, and may become robust after the imaginal molt.