Reproductive Arrest Research Articles

Aminergic Signaling Controls Ovarian Dormancy in Drosophila

In response to adverse environmental conditions many organisms from nematodes to mammals deploy a dormancy strategy, causing states of developmental or reproductive arrest that enhance somatic maintenance and survival ability at the expense of growth or reproduction. Dormancy regulation has been studied in C. elegans and in several insects, but how neurosensory mechanisms act to relay environmental cues to the endocrine system in order to induce dormancy remains unclear. Here we examine this fundamental question by genetically manipulating aminergic neurotransmitter signaling in Drosophila melanogaster. We find that both serotonin and dopamine enhance adult ovarian dormancy, while the downregulation of their respective signaling pathways in endocrine cells or tissues (insulin producing cells, fat body, corpus allatum) reduces dormancy. In contrast, octopamine signaling antagonizes dormancy. Our findings enhance our understanding of the ability of organisms to cope with unfavorable environments and illuminate some of the relevant signaling pathways.

Stress-induced reproductive arrest in Drosophila occurs through ETH deficiency-mediated suppression of oogenesis and ovulation

BackgroundEnvironmental stressors induce changes in endocrine state, leading to energy re-allocation from reproduction to survival. Female Drosophila melanogaster respond to thermal and nutrient stressors by arresting egg production through elevation of the steroid hormone ecdysone. However, the mechanisms through which this reproductive arrest occurs are not well understood.ResultsHere we report that stress-induced elevation of ecdysone is accompanied by decreased levels of ecdysis triggering hormone (ETH). Depressed levels of circulating ETH lead to attenuated activity of its targets, including juvenile hormone-producing corpus allatum and, as we describe here for the first time, octopaminergic neurons of the oviduct. Elevation of steroid thereby results in arrested oogenesis, reduced octopaminergic input to the reproductive tract, and consequent suppression of ovulation. ETH mitigates heat or nutritional stress-induced attenuation of fecundity, which suggests that its deficiency is critical to reproductive adaptability.ConclusionsOur findings indicate that, as a dual regulator of octopamine and juvenile hormone release, ETH provides a link between stress-induced elevation of ecdysone levels and consequent reduction in fecundity.

Diapause in Drosophila melanogaster – Photoperiodicity, cold tolerance and metabolites

Unlike many insects where photoperiod per se induces diapause, reproductive arrest in Drosophila melanogaster adult females is observed at colder temperatures and can be enhanced by shorter photoperiods. Traditional experimental protocols raise flies at 25 °C from the larval stage and then the adults are placed at 12 °C for between 12 and 28 days. After 12 days diapause levels are usually higher than at 28 days, suggesting that the flies are in a cold induced quiescence, rather than a true diapause. By raising flies at more realistic lower temperatures, we observe quite dramatic and counter-intuitive effects on diapause, whose levels nevertheless correlate with various indices of cryoprotectant metabolites as well as resistance to chill shock. We also observe that photoperiodic effects are minimised when very small temperature oscillations associated with the light-dark incubator cycles are neutralised. Our results suggest that the reported photoperiodic component of fly diapause, at least in these strains, is mostly due to thermoperiodic rather than photoperiodic stimuli. In addition, the metabolite and chill shock analyses reveal that even by 12 days, flies are entering a state that is resistant to environmental stresses.

Influence of sexual behavior of Dorper rams treated with glutamate and/or testosterone on reproductive performance of anovulatory ewes

The aim of this study was to determine if exogenous administration of glutamate and (or) testosterone to male rams during the season of reproductive arrest is able to re-activate male sexual behavior and, later on, to promote through the male effect, both sexual and reproductive competence of anovulatory nulliparous ewes. Therefore, an experiment was performed under long-day photoperiods (spring; photo-reproductive arrest, 26°N). Dorper rams were randomly divided into four homogeneous experimental groups (n = 5 males each) regarding live weight (LW), body condition score (BCS), scrotal circumference (SC) and odor intensity (OI). Then, groups were treated with: i) GG (7 mg kg−1 LW of glutamate, every 4d × 30d, im.), ii) GGT (7 mg kg−1 LW of glutamate every 4d × 30d im + 25 mg of testosterone propionate, every 3d × 15d, im.), iii) GT (25 mg of testosterone propionate every 3d × 15d, im, and iv) GC (1 mL of saline, every 4d × 30d, im.). Thereafter, Dorper rams, (n = 4 per group) were selected and exposed to Dorper anovulatory-nulliparous ewes divided in four groups (n = 14 ewes each), and all the appetitive (ASB) and consummatory (CSB) sexual behaviors and indicators of sexual rest (ISR) were registered during the first 48 h of this male-to-females contact. Thereafter, males continued the male-to-female contact for another 8 d, in order to quantify the ewe's sexual and reproductive response through the male effect. During the sexual behavior tests, the GGT rams showed the highest ASB + CSB frequencies (P < 0.05) followed by the GG-rams with the lowest frequencies showed by the GC and GT groups. While the highest ISR behaviors were shown by the GG and GGT groups (P < 0.05) followed by the GC and GT-rams, no differences occurred regarding LW, BCS, and SC along the experimental breeding, with the largest (P < 0.05) OI shown by the GGT-rams and the lowest value observed in the control rams. Regarding the reproductive response of the nulliparous-anovulatory ewes exposed to treated rams, the glutamate + testosterone treated males induced not only an increased (P < 0.05) ovulatory activity and faster (P < 0.05) estrus response but an augmented (P < 0.05) pregnancy rate of the previously anovulatory ewes. To conclude, whereas the GG and GGT treatments generated encouraging sexual and reproductive outcomes, our results are also thought-provoking from a comparative sexual behavior perspective while may embrace potential applications to other animal industries.

Dynamics of ovarian maturation throughout the reproductive cycle of the Neotropical cichlid fishCichlasomadimerus(Teleostei, Cichliformes)

In this study, we analyzed gene expression profiles, plasma steroids concentrations, and gonadal morphology throughout the reproductive cycle of female Cichlasoma dimerus (Heckel, 1840), a monogamous cichlid fish exhibiting social hierarchies. Fish were analyzed at six phases encompassing their annual cycle, namely resting (during the nonreproductive period), prespawning, 30 h post spawning, 4 days post spawning, 10 days post spawning, and subordinate (during the reproductive period). The histological and histomorphometric analysis showed that C. dimerus exhibits asynchronous ovarian development. Similar to resting females, subordinate females showed low gonadosomatic index, reduced expression levels of vitellogenin (vtgAb), zona pellucida (zpB), gonadal aromatase (cyp19a1A), and low concentrations of plasma sex steroids, thus indicating that social intimidation by dominant conspecifics elicited reproductive arrest. In reproductively active females, a direct positive correlation between plasma estradiol, vtgAb expression, percentage of late vitellogenic oocytes, and gonadosomatic index was observed. These parameters were maximal at the prespawning phase, decreased at 30 h post spawning and 4 days post spawning, and then reached a peak at 10 days post spawning. Our results indicate that female C. dimerus become spawning capable after 10 days post spawning, coincidently with the shortest time interval between successive spawns recorded in captivity.

Tracing the origin of the early wet-season Anopheles coluzzii in the Sahel.

In arid environments, the source of the malaria mosquito populations that re‐establish soon after first rains remains a puzzle and alternative explanations have been proposed. Using genetic data, we evaluated whether the early rainy season (RS) population of Anopheles coluzzii is descended from the preceding late RS generation at the same locality, consistent with dry season (DS) dormancy (aestivation), or from migrants from distant locations. Distinct predictions derived from these two hypotheses were assessed, based on variation in 738 SNPs in eleven A. coluzzii samples, including seven samples spanning 2 years in a Sahelian village. As predicted by the “local origin under aestivation hypothesis,” temporal samples from the late RS and those collected after the first rain of the following RS were clustered together, while larger genetic distances were found among samples spanning the RS. Likewise, multilocus genotype composition of samples from the end of the RS was similar across samples until the following RS, unlike samples that spanned the RS. Consistent with reproductive arrest during the DS, no genetic drift was detected between samples taken over that period, despite encompassing extreme population minima, whereas it was detected between samples spanning the RS. Accordingly, the variance in allele frequency increased with time over the RS, but not over the DS. However, not all the results agreed with aestivation. Large genetic distances separated samples taken a year apart, and during the first year, within‐sample genetic diversity declined and increased back during the late RS, suggesting a bottleneck followed by migration. The decline of genetic diversity followed by a mass distribution of insecticide‐treated nets was accompanied by a reduced mosquito density and a rise in the mutation conferring resistance to pyrethroids, indicating a bottleneck due to insecticidal selection. Overall, our results support aestivation in A. coluzzii during the DS that is accompanied by long‐distance migration in the late RS.

The role of circadian clock genes in the photoperiodic timer of the linden bug Pyrrhocoris apterus during the nymphal stage

AbstractMany insects survive seasonal adversities during diapause, a form of programmed developmental and metabolic arrest. Photoperiodically regulated entry into diapause allows multivoltine insect species to optimize the number of generations. The molecular mechanism of the photoperiodic timer is unknown in insects. In the present study, we take advantage of the robust reproductive diapause response in the linden bug Pyrrhocoris apterus and explore the fifth‐instar nymphal stage, which is the most photoperiod‐sensitive stage. The nymphs display daily changes in locomotor activity during short days; this differs from the activity observed during long days. We find evidence of cyclical expression of the circadian clock genes, per and cyc, in nymphal heads; in addition, per expression is also photoperiod‐dependent. The RNA interference‐mediated knockdown of the two circadian clock genes, Clk and cyc, during the nymphal stage results in reproductive arrest in adult females. Furthermore, Clk and cyc knockdown induces the expression of the storage protein hexamerin in the fat body, whereas the expression of vitellogenin diminishes. Taken together, these data support the involvement of circadian clock genes in photoperiodic timer and/or diapause induction.

Is diapause an ancient adaptation in Drosophila?

D. melanogaster enters a state of reproductive arrest when exposed to low temperatures (12°C) and shorter photoperiods. A number of studies have suggested that diapause has recently evolved in European D. melanogaster populations, that it is not present in the sibling species D. simulans, that it is non-photoperiodic in American D. melanogaster populations, and that it spontaneously terminates after 6–8weeks. We have studied the overwintering phenotype under different conditions and observe that American, European and, surprisingly, African D. melanogaster populations can show photoperiodic diapause, as can European, but not African D. simulans. Surprisingly other Drosophila species from pan-tropical regions can also show significant levels of photoperiodic diapause. We observe that spontaneous termination of diapause after a few weeks can be largely avoided with a more realistic winter simulation for D. melanogaster, but not D. simulans. Examining metabolite accumulation during diapause reveals that the shallow diapause of D. melanogaster has similar features to that of other more robustly-diapausing species. Our results suggest that diapause may be an ancient character that emerged in the tropics to resist unfavourable seasonal conditions and which has been enhanced during D. melanogaster’s colonisation of temperate regions. Our results also highlight how different methodologies to quantify diapause can lead to apparently conflicting results that we believe can now largely be resolved.

The Insulin-Like Proteins dILPs-2/5 Determine Diapause Inducibility in Drosophila.

Diapause is an actively induced dormancy that has evolved in Metazoa to resist environmental stresses. In temperate regions, many diapausing insects overwinter at low temperatures by blocking embryonic, larval or adult development. Despite its Afro-tropical origin, Drosophila melanogaster migrated to temperate regions of Asia and Europe where females overwinter as adults by arresting gonadal development (reproductive diapause) at temperatures <13°C. Recent work in D. melanogaster has implicated the developmental hormones dILPs-2 and/or dILP3, and dILP5, homologues of vertebrate insulin/insulin-like growth factors (IGFs), in reproductive arrest. However, polymorphisms in timeless (tim) and couch potato (cpo) dramatically affect diapause inducibility and these dILP experiments could not exclude this common genetic variation contributing to the diapause phenotype. Here, we apply an extensive genetic dissection of the insulin signaling pathway which allows us to see both enhancements and reductions in egg development that are independent of tim and cpo variations. We show that a number of manipulations dramatically enhance diapause to ~100%. These include ablating, or reducing the excitability of the insulin-producing cells (IPCs) that express dILPs-2,3,5 employing the dilp2,3,5-/- triple mutant, desensitizing insulin signaling using a chico mutation, or inhibiting dILP2 and 5 in the hemolymph by over-expressing Imaginal Morphogenesis Protein-Late 2 (Imp-L2). In addition, triple mutant dilp2,3,5-/- females maintain high levels of diapause even when temperatures are raised in adulthood to 19°C. However at 22°C, these females all show egg development revealing that the effects are conditional on temperature and not a general female sterility. In contrast, over-expression of dilps-2/5 or enhancing IPC excitability, led to levels of ovarian arrest that approached zero, underscoring dILPs-2 and 5 as key antagonists of diapause.

Developmental Acclimation of Drosophila suzukii (Diptera: Drosophilidae) and Its Effect on Diapause and Winter Stress Tolerance.

We investigated the influence of developmental conditions on adult morphology, reproductive arrest, and winter stress tolerance of the invasive pest of small fruit, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae). Cooler rearing temperatures (15 °C) resulted in larger, darker "winter morph" (WM) adults than "summer morph" flies reared at optimal temperatures (25 °C). Abdominal pigmentation scores and body size measurements of laboratory-reared WMs were similar to those of D. suzukii females captured in late autumn in Geneva, NY. We evaluated reproductive diapause and cold hardiness in live-captured D. suzukii WMs as well as WMs reared in the laboratory from egg to adult under four developmental conditions: static cool temperatures (SWM; 15 °C, 12:12 h L:D), fluctuating temperatures (FWM; 20 °C L: 10 °C D, 12:12 h L:D), and static cool temperatures (15 °C, 12:12 h L:D) followed by posteclosion chilling (CWM; 10 °C) under short-day (SD; 12:12 h L:D) or long-day photoperiods (LD; 16:8 h L:D). Live-captured D. suzukii WMs and CWMs had longer preoviposition times than newly eclosed summer morph adults, indicating a reproductive diapause that was not observed in SWMs or FWMs. Additionally, recovery after acute freeze stress was not different between CWM-SD females and live captured WM females. More 7-d-old CWMs survived 0, -1, or - 3 °C freeze stress than summer morph adults, and more CWM-SD adults survived -3 °C freeze stress than CWM-LD adults. Survival after -3 °C freeze stress was significantly higher in diapausing, CWMs than nondiapausing SWMs and FWMs.

Reproductive arrest and stress resistance in winter-acclimated Drosophila suzukii

Overwintering insects must survive the multiple-stress environment of winter, which includes low temperatures, reduced food and water availability, and cold-active pathogens. Many insects overwinter in diapause, a developmental arrest associated with high stress tolerance. Drosophila suzukii (Diptera: Drosophilidae), spotted wing drosophila, is an invasive agricultural pest worldwide. Its ability to overwinter and therefore establish in temperate regions could have severe implications for fruit crop industries. We demonstrate here that laboratory populations of Canadian D. suzukii larvae reared under short-day, low temperature, conditions develop into dark ‘winter morph’ adults similar to those reported globally from field captures, and observed by us in southern Ontario, Canada. These winter-acclimated adults have delayed reproductive maturity, enhanced cold tolerance, and can remain active at low temperatures, although they do not have the increased desiccation tolerance or survival of fungal pathogen challenges that might be expected from a more heavily melanised cuticle. Winter-acclimated female D. suzukii have underdeveloped ovaries and altered transcript levels of several genes associated with reproduction and stress. While superficially indicative of reproductive diapause, the delayed reproductive maturity of winter-acclimated D. suzukii appears to be temperature-dependent, not regulated by photoperiod, and is thus unlikely to be ‘true’ diapause. The traits of this ‘winter morph’, however, likely facilitate overwintering in southern Canada, and have probably contributed to the global success of this fly as an invasive species.

Enhanced fertility and chill tolerance after cold-induced reproductive arrest in females of temperate species of the Drosophila buzzatii complex.

Long-term exposure to low temperatures during adult maturation might decrease fertility after cold recovery as a consequence of carry-over effects on reproductive tissues. This pattern should be more pronounced in tropical than in temperate species as protective mechanisms against chilling injuries are expected to be more effective in the latter. We initially determined the lower thermal thresholds to induce ovarian maturation in four closely related Drosophila species, two inhabiting temperate regions and the other two tropical areas of South America. As expected, only temperate species regularly experience cold-inducing conditions for reproductive arrest during winter in their natural environment. Subsequently, we exposed reproductively arrested and mature females to cold-inducing conditions for reproductive arrest over a long period. Following cold exposure, tropical species exhibited a dramatic fertility decline, irrespective of reproductive status. In contrast, not only were temperate females fecund and fertile but also fertility was superior in females that underwent cold-induced reproductive arrest, suggesting that it might act as a protecting mechanism ensuring fertility after cold recovery. Based on these findings, we decided to evaluate the extent to which reproductive status affects cold tolerance and energy metabolism at low temperature. We found a lower metabolic rate and a higher cold tolerance in reproductively arrested females, although only temperate species attained high levels of chill tolerance. These findings highlight the role of cold-induced reproductive arrest as part of an integrated mechanism of cold adaptation that could potentially contribute to the spread of temperate species into higher latitudes or altitudes.

Can heterothermy facilitate the colonization of new habitats?

Abstract Increasing evidence shows that torpor in mammals is not only an effective adaptation for surviving predictable seasonal harsh conditions but is also employed as a response to acute emergency situations. This finding leads to the hypothesis that the ability to become heterothermic can also facilitate the colonization of new habitats, when mammals have to cross unsuitable, fragmented and/or food‐scarce landscapes. This hypothesis was first suggested in the context of the colonization of Madagascar, and it has never been evaluated in detail. In this review, we discuss the potential of heterothermy to affect colonization possibilities in general and the colonization of Madagascar in particular. We list several points supporting the hypothesis and contend that, unless colonization events coincidentally take place during the hibernation period, spontaneous daily torpor is probably more instrumental for surviving unexpected and severe conditions on a raft or when crossing harsh landscapes than seasonally occurring hibernation. We further propose that torpor can positively affect the establishment of a founder population by enabling reproductive arrest phases until conditions improve. Moreover, if viable offspring are born after arrival, this might abolish the absolute necessity of encountering individuals of the opposite sex. Altogether, it seems therefore consequential that heterothermic mammals with the ability to enter daily torpor are the most likely to arrive in a new habitat in a body condition that allows successful settlement and reproduction. Together with the finding that all recent taxa of terrestrial, non‐volant Malagasy mammals or their close relatives include representatives that use heterothermy, this suggests that torpor might indeed have facilitated the colonization of Madagascar.

Secretory activity of mandibular organ fluctuates in response to reproductive season of the field crab Paratelphusa sp. (Brachyura; Decapoda): an ultrastructural study

Abstract Background Mandibular organ (MO) in decapods is suggested to play regulatory role in reproduction, in few species; however, MO is considered to control growth. The present study addresses this question by an ultrastructural study on the MO of the field crab, Paratelphusa sp. Our sampling for consecutive years (2008 to 2012) revealed that Paratelphusa sp. devotes July to October for reproduction, judged by the occurrence of growing ovaries and the berried females. From November to the succeeding June, the females are in a state of reproductive arrest (non-reproductive period); ovaries during this season would appear as white bands with no signs of yolk deposition. Results Morphologically, MO of Paratelphusa sp. is positioned posterior to the mandibles and is in close apposition with the distal end of the mandibular apodeme. MO of Paratelphusa sp. exhibited significant levels (t = 8.097, P &lt; 0.0001, N = 10) of season-dependent size variations. Our electron microscopic observations reveal that the MO is highly secretory during the reproductive period, evidenced by the occurrence of sacculated Golgi bodies having dense inclusions, several mitochondria with tubular cristae, and extensive networks of SER and rough endoplasmic reticulum (RER). During the non-reproductive period, however, the MO is least active; RER, the mitochondria, and the Golgi are only sparsely seen. Interestingly, the plasma membrane exhibits a highly convoluted appearance all the way through the non-reproductive period. Conclusions The present study reveals that the secretory activity of MO of Paratelphusa sp. is entrained with reproductive activity. The existence of a high correlation between MO secretory activity and ovarian growth implicates the former’s role in reproduction.

Overwintering Biology of &lt;I&gt;Culex&lt;/I&gt; (Diptera: Culicidae) Mosquitoes in the Sacramento Valley of California

At temperate latitudes, Culex (Diptera: Culicidae) mosquitoes typically overwinter as adult females in reproductive arrest and also may serve as reservoir hosts for arboviruses when cold temperatures arrest viral replication. To evaluate their role in the persistence of West Nile virus (WNV) in the Sacramento Valley of California, the induction and termination of diapause were investigated for members of the Culex pipiens (L.) complex, Culex tarsalis Coquillett, and Culex stigmatosoma Dyar under field, seminatural, and experimental conditions. All Culex spp. remained vagile throughout winter, enabling the collection of 3,174 females and 1,706 males from diverse habitats during the winters of 2010-2012. Overwintering strategies included both quiescence and diapause. In addition, Cx. pipiens form molestus Forskäl females remained reproductively active in both underground and aboveground habitats. Some blood-fed, gravid, and parous Cx. tarsalis and Cx. pipiens complex females were collected throughout the winter period. Under both field and experimental conditions, Cx. tarsalis and Cx. stigmatosoma females exposed to autumnal conditions arrested primary follicular maturation at previtellogenic stage I, with primary to secondary follicular ratios <1.5 (indicative of a hormonally induced diapause). In contrast, most Cx. pipiens complex females did not enter reproductive diapause and ovarian follicles matured to >or=stage I-II (host-seeking arrest) or were found in various stages of degeneration. Diapause was initiated in the majority of Cx. tarsalis and Cx. stigmatosoma females by mid-late October and was terminated after the winter solstice, but host-seeking seemed limited by temperature. An accrual of 97.52 +/- 30.7 and 162.85 +/- 79.3 degree-days after the winter solstice was estimated to be necessary for diapause termination in Cx. tarsalis under field and seminatural conditions, respectively. An increase in the proportion of blood-fed Culex females in resting collections occurred concurrently with diapause termination in field populations based on ovarian morphometrics. WNV RNA was detected in one pool of 18 males and in a single blood-fed female Cx. tarsalis collected during winter. Therefore, both vertically and horizontally infected Culex females may persist through winter and possibly transmit WNV after diapause termination in late winter or early spring in the Sacramento Valley of California.

Circannual Variation in Thyroid Hormone Deiodinases in a Short‐Day Breeder

At temperate latitudes, many mammals and birds show internally timed, long-term changes in seasonal physiology, synchronised to the seasons by changing day length (photoperiod). Photoperiodic control of thyroid hormone levels in the hypothalamus dictates the timing. This is effected through reciprocal regulation of thyroid hormone deiodinase gene expression. The local synthesis of type 2 deiodinase (Dio2) promotes triiodothyronine (T3) production and summer biology, whereas type 3 deiodinase (Dio3) promotes T3 degradation and winter biology. In the present study, we investigated the extent to which the hypothalamic expression of Dio2 and Dio3 is circannually regulated in the Soay sheep, a short-day breeding mammal. Male sheep were exposed to a long photoperiod (LP; 16 : 24 h light/dark cycle) or a short photoperiod (SP; 8 : 24 h light/dark cycle), for up to 28 weeks to establish four different endocrine states: (i) LP animals in a spring/summer-like state of reproductive arrest; (ii) LP refractory (LPR) animals showing spontaneous reproductive reactivation; (iii) SP animals showing autumn/winter-like reproductive activation; and (iv) SP refractory (SPR) animals showing spontaneous reproductive arrest. A complex pattern of hypothalamic Dio2 and Dio3 expression was observed, revealing distinctive photoperiod-driven and internally timed effects for both genes. The patterns of expression differed both spatially and temporally, with phases of peak Dio2 expression in the median eminence and tuberoinfundibular sulcus, as well as in the paraventricular zone (PVZ) (maximal under LP), whereas Dio3 expression was always confined to the PVZ (maximal under SP). These effects likely reflect the distinct roles of these enzymes in the localised control of hypothalamic T3 levels. The spontaneous decline in Dio2 and spontaneous increase in Dio3 in LPR animals occurred with a corresponding decline in thyroid-stimulating hormone β expression in the neighbouring pars tuberalis (PT), although this relationship did not hold for the corresponding Dio2 increase/Dio3 decrease seen in SPR animals. We conclude that internally timed and spatially regulated changes in Dio2 and Dio3 expression may drive the cycling between breeding and nonbreeding states in long-lived seasonal species, and may be either PT-dependent or PT-independent at different phases of the circannual cycle.

Autonomous regulation of the insect gut by circadian genes acting downstream of juvenile hormone signaling

In temperate regions, the shortening day length informs many insect species to prepare for winter by inducing diapause. The adult diapause of the linden bug, Pyrrhocoris apterus, involves a reproductive arrest accompanied by energy storage, reduction of metabolic needs, and preparation to withstand low temperatures. By contrast, nondiapause animals direct nutrient energy to muscle activity and reproduction. The photoperiod-dependent switch from diapause to reproduction is systemically transmitted throughout the organism by juvenile hormone (JH). Here, we show that, at the organ-autonomous level of the insect gut, the decision between reproduction and diapause relies on an interaction between JH signaling and circadian clock genes acting independently of the daily cycle. The JH receptor Methoprene-tolerant and the circadian proteins Clock and Cycle are all required in the gut to activate the Par domain protein 1 gene during reproduction and to simultaneously suppress a mammalian-type cryptochrome 2 gene that promotes the diapause program. A nonperiodic, organ-autonomous feedback between Par domain protein 1 and Cryptochrome 2 then orchestrates expression of downstream genes that mark the diapause vs. reproductive states of the gut. These results show that hormonal signaling through Methoprene-tolerant and circadian proteins controls gut-specific gene activity that is independent of circadian oscillations but differs between reproductive and diapausing animals.

Relationship between body colour, feeding, and reproductive arrest under short-day development in Tetranychus pueraricola (Acari: Tetranychidae)

In Tetranychus spider mites (Acari: Tetranychidae), diapausing females have a conspicuous orange body colour, which is used as an indicator of diapause induction in many laboratory studies. However, to which extent body colour reflects reproductive activity is scarcely investigated. In this study, we investigated the relationship between body colour, reproductive arrest, and food intake in the inbred strain of T. pueraricola individually reared at 20 °C with a 10:14 h light: dark photoperiod. Our results showed that (1) body colour is a good indicator of reproductive arrest 11 days after adult emergence but does not completely reflect reproductive status at an earlier age; (2) even orange females intermittently feed, and the arrest of feeding comes after the change in body colour; and (3) reproducing females have a higher risk of death than non-reproducing females. These results suggest that measurement of diapause incidence by body colour alone may miss the variation in reproductive status in early adult life.

Do long- and short-winged adult females of the bug Pyrrhocoris apterus (Heteroptera: Pyrrhocoridae) differ in lifespan and reproductive capacity?

In the present study we tested whether long-winged (macropterous) and short-winged (brachypterous) adult females of the firebug Pyrrhocoris apterus (L.) differ in their reproductive capacity and length of life. The following seven physiological markers were measured in these females: lengths of the pre-oviposition period (pre-OP), inter-ovipositon period (inter-OP) and post-oviposition period (post-OP), and the mean number of eggs per batch, total number of eggs, mean total number of egg batches laid and lifespan of the females. The results showed that macropterous and brachypterous females significantly differed in the length of the pre-OP, which was significantly shorter in brachypterous (7.95 ± 1.75 days) than in macropterous females (26.84 ± 9.86 days), but there was no significant difference between the lengths of the inter-OP in brachypterous (4.00-8.79 days) and macropterous (3.00-9.89 days) females. In contrast the length of the post-OP was significantly longer in brachypterous (48.23 ± 30.95 days) than in macropterous females (35.02 ± 17.32 days). Except for the 2nd and 3rd egg batches there was no significant difference between the average numbers of eggs in the other egg batches laid by females of the two wing morphs. Total number of eggs laid by macropterous females during their whole lifespan was not significantly lower (326.47 ± 155.65 eggs) than by brachypterous females (382.82 ± 207.52 eggs), but associated with the lower number of egg batches laid by macropterous (13) than by brachypterous females (19). However, there was no significant difference in the longevity of brachypterous (95.43 ± 41.21 days) and macropterous (93.40 ± 21.18 days) females. The relationship of these results to the reproductive arrest, inactivity of the endocrine gland, corpus allatum, and different levels of adipokinetic hormone in macropterous females is discussed.

Defining behavioral and molecular differences between summer and migratory monarch butterflies

BackgroundIn the fall, Eastern North American monarch butterflies (Danaus plexippus) undergo a magnificent long-range migration. In contrast to spring and summer butterflies, fall migrants are juvenile hormone deficient, which leads to reproductive arrest and increased longevity. Migrants also use a time-compensated sun compass to help them navigate in the south/southwesterly direction en route for Mexico. Central issues in this area are defining the relationship between juvenile hormone status and oriented flight, critical features that differentiate summer monarchs from fall migrants, and identifying molecular correlates of behavioral state.ResultsHere we show that increasing juvenile hormone activity to induce summer-like reproductive development in fall migrants does not alter directional flight behavior or its time-compensated orientation, as monitored in a flight simulator. Reproductive summer butterflies, in contrast, uniformly fail to exhibit directional, oriented flight. To define molecular correlates of behavioral state, we used microarray analysis of 9417 unique cDNA sequences. Gene expression profiles reveal a suite of 40 genes whose differential expression in brain correlates with oriented flight behavior in individual migrants, independent of juvenile hormone activity, thereby molecularly separating fall migrants from summer butterflies. Intriguing genes that are differentially regulated include the clock gene vrille and the locomotion-relevant tyramine beta hydroxylase gene. In addition, several differentially regulated genes (37.5% of total) are not annotated. We also identified 23 juvenile hormone-dependent genes in brain, which separate reproductive from non-reproductive monarchs; genes involved in longevity, fatty acid metabolism, and innate immunity are upregulated in non-reproductive (juvenile-hormone deficient) migrants.ConclusionThe results link key behavioral traits with gene expression profiles in brain that differentiate migratory from summer butterflies and thus show that seasonal changes in genomic function help define the migratory state.