Sexual dimorphism of sleep regulated by juvenile hormone signaling in Drosophila

Binbin Wu; Lingling Ma; Enyan Zhang; Juan Du; Suning Liu; Jeffrey Price; Sheng Li; Zhangwu Zhao

doi:10.1371/journal.pgen.1007318

Abstract

Sexually dimorphic phenotypes are a universal phenomenon in animals. In the model animal fruit fly Drosophila, males and females exhibit long- and short-sleep phenotypes, respectively. However, the mechanism is still a mystery. In this study, we showed that juvenile hormone (JH) is involved in regulation of sexually dimorphic sleep in Drosophila, in which gain of JH function enlarges differences of the dimorphic sleep phenotype with higher sleep in males and lower sleep in females, while loss of JH function blurs these differences and results in feminization of male sleep and masculinization of female sleep. Further studies indicate that germ cell-expressed (GCE), one of the JH receptors, mediates the response in the JH pathway because the sexually dimorphic sleep phenotypes cannot be rescued by JH hormone in a gce deletion mutant. The JH-GCE regulated sleep dimorphism is generated through the sex differentiation-related genes -fruitless (fru) and doublesex (dsx) in males and sex-lethal (sxl), transformer (tra) and doublesex (dsx) in females. These are the “switch” genes that separately control the sleep pattern in males and females. Moreover, analysis of sleep deprivation and circadian behaviors showed that the sexually dimorphic sleep induced by JH signals is a change of sleep drive and independent of the circadian clock. Furthermore, we found that JH seems to also play an unanticipated role in antagonism of an aging-induced sleep decrease in male flies. Taken together, these results indicate that the JH signal pathway is critical for maintenance of sexually dimorphic sleep by regulating sex-relevant genes.

Author summary

Sleep is a very important biological behavior in all animals and takes up around one third of the lifespan in many animals. In both insects and mammals (including humans), sleep differences between male and female (sexually dimorphic sleep) have been described over the past decades. However, its internal regulation mechanism is still unclear. The fruit fly Drosophila melanogaster, sharing most sleep characteristics with humans, has been used for sleep studies as a powerful model for genetic analysis. In this study, we reported that Juvenile hormone (JH) induces completely different sleep effects between males and females with higher sleep in males and lower sleep in females, while loss of JH function blurs these differences and results in feminization of male sleep and masculinization of female sleep. Further studies indicate that the sexual dimorphism of sleep is generated through the sex differentiation-related genes regulated by JH and its receptor GCE (germ cell-expressed) signaling. Furthermore, we found that JH seems to also play an unanticipated role in aging-induced sleep changes.

Citation: Wu B, Ma L, Zhang E, Du J, Liu S, Price J, et al. (2018) Sexual dimorphism of sleep regulated by juvenile hormone signaling in Drosophila. PLoS Genet 14(4): e1007318. https://doi.org/10.1371/journal.pgen.1007318

Editor: Paul H. Taghert, Washington University in Saint Louis School of Medicine, UNITED STATES

Received: September 6, 2017; Accepted: March 19, 2018; Published: April 4, 2018

Copyright: © 2018 Wu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the paper and its Supporting Information files.

Funding: The work was funded by grants from the National Nature Science Foundation of China (Grant numbers 31572317 and 31730076) to ZZ. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Introduction

Sleep is a general phenomenon identified in many species and is usually accompanied with various sex-specific properties. For instance, in human beings, women have higher sleep spindle number and density than men[1]. Also there are sex differences in responses of male and female rats to sleep deprivation[2]. In recent years, the Drosophila genetic model system for sleep research has been shown to share all the key characteristics of mammalian sleep[3,4,5]. Similar to a variety of organisms, sex-specific properties in sleep pattern are present in fruit flies. Previous studies reported that there is a large sex difference in the total sleep amounts during the daylight hours, in which female sleep is only 40% of male sleep in D. melanogaster[6].To address that phenomenon, specific neurons[7,8] and non-neural factors [8] relevant to sexually dimorphic sleep have been discussed. Other types of sexual dimorphism (e.g., stress resistance, feeding, and physical characteristics) mediated by insulin, dopamine and Juvenile Hormone (JH) may participate in the relevant regulatory circuits[9,10], but their specific interactions and the molecular mechanism are still unclear. Here, we examine the role of JH in the control of a sexually dimorphic sleep phenotype.

Juvenile hormones are a group of acyclic sesquiterpenoids that regulate insect physiological processes, such as development, reproduction and diapause[11]. JHs are synthesized primarily in the corpora allata (CA), a pair of endocrine glands with neural connections to the brain[12]. The pars intercerebralis (PI) has been recently identified as a structure of neuroendocrine regulation for sleep and wakefulness in Drosophila[13]. It has been suggested that at least part of the CA is derived from the PIa/c (PIa: located around the medial lobes of the mushroom body; PIc: located dorsal to the central complex)[14]. Furthermore, JH could serve as a molecular link between PI neurons and the corpus cardiacum—copora allata(CC-CA) gland[15]. Previous studies mapped the PI area of the brain which feminizes male locomotor activity through up-regulated expression of the sex-determination gene transformer (tra)[16].

In D. melanogaster, the presence of XX (or XY) sex chromosome constitution promotes female (or male) somatic development through a sex determination hierarchy[17]. The binary switch gene sex-lethal (sxl) regulated by the X chromosome counting system controls sexual identity in Drosophila. When SXL is on, it imposes female development, whereas male development proceeds when it is off[18]. Transformer (tra), fruitless (fru) and doublesex (dsx) act downstream of sxl to control sexual development in Drosophila. TRA regulates female-specific alternative splicing of dsx to encode DSX^F protein. Without TRA, male-specific alternative splicing of dsx pre-mRNA occurs, and this transcript encodes DSX^M protein[19]. Fru, a pleiotropic gene which lies at the bottom of the sex-determination hierarchy, controls male sexual behavior and is essential for viability in Drosophila[20]. Indeed, these genes products control sexual differentiation of the organism body and also contribute to relevant behavioral and neural development[21].

Biosynthetic activity in the CA is considered to be a major factor in the regulation of JH titer[22,23]. The JH biosynthetic pathway mainly includes two steps: the generation of farnesyl diphosphate (FPP) via the mevalonate pathway in early steps, and then conversion of farnesoic acid (FA) generated from FPP to active JH in later steps[11], in which many enzymes have been found to be responsible for the biosynthesis of JH, such as HMG-CoA synthase (HMGS), HMG-CoA reductase (HMGCR), phosphomevalonate kinase (MevPK), and JH acid O-methyltransferase (JHAMT)[24,25]. Among these, the enzyme JHAMT expressed in the CA activates the JH biosynthesis pathway in the last step and is considered the key regulator in JH biosynthesis. In D. melanogaster, the JHAMT expression pattern is consistent with changes of the JH titer, and recombinant DmJHAMT protein can produce JHs with the presence of its substrates[23]. In addition, the rate of JH in vitro biosynthesis by brain-ring gland complex is higher when JHAMT is over-expressed in the CA[26]. In Bombyx mori, jhamt mRNA level is consistent with JH titer, and the transcriptional suppression of jhamt gene is crucial for down-regulation of JH biosynthesis[11,25]. Therefore, JHAMT is a useful and important index for JH. In this study, we explored the role of JH and its signal pathway on sleep regulation through a genetically transgenic system.

Results

Effects of JH on sleep

To compare sleep-wake profiles between both sexes, we monitored hundreds of wild type flies. Results showed that sleep of males during daytime was much higher than that of females (Fig 1A). Transcriptomic analysis of sleep-deprived and non-deprived rats has shown that sleep–wakefulness states affect gene expression[27]. Thus, we performed a sleep deprivation assay by forcing flies of wild-type w¹¹¹⁸ to be awake all night using mechanical vibration, and surprisingly found that the changes of jhamt transcript levels exhibited two opposite tendencies in the male and female flies after 12 h sleep deprivation (Fig 1B). To further elucidate the function of endogenous JH signaling on the regulation of sleep in flies, we used a tissue-specific driver (Aug21-gal4) to over-express JHAMT—a key enzyme synthesizing JH in the CA cells[26,28]. Over-expression was confirmed by immunofluorescence with anti-DmJHAMT antibody (Fig 1C). Results showed that flies with up-regulated JHAMT exhibited enhanced sleep differences between both sexes, in which the daytime sleep amount in males showed a significant increase but a significant decrease in females(Fig 1D and 1E), primarily caused by sleep episode duration (Fig 1F). Furthermore, in order to eliminate the possibility that abnormality of metamorphosis development would contribute to non-specific effects on sleep, we applied three different concentrations of exogenous pyriproxyfen–a juvenile hormone analog (JHA)- to adult wild-type flies, and then sleep behavior was monitored. Results showed that flies with JHA application exhibited similar sexually dimorphic sleep responses like those above with genetic gain of JHAMT function, with 0.1mM JHA as an effective concentration and regulation of sleep showing dose-dependency(Fig 1G–1I).

Download:

Fig 1.

Effects of gain of JH function on sleep behaviors. (A) Sleep patterns of male and female flies; the gray represents w¹¹¹⁸ female, and the blue represents w¹¹¹⁸ male. Individual plots for multiple females and males are shown, in which one plot trace line represents a repeat with 16–32 flies. Tabulated data represent mean±SEM (n = 112–145).***P<0.001 determined by Student’s t test. (B) Jhamt relative expression in w¹¹¹⁸ males and females after sleep deprivation. Data represent mean±SEM. *P<0.05 determined by Student’s t test. (C) Aug21-gal4 was used to drive JHAMT over-expression in the corpora allata cells. The blue represents DAPI, the red represents anti-JHAMT, and the third column represents co-location of DAPI and anti-JHAMT. (D) Sleep trace graph of flies over-expressing jhamt with Aug21-gal4. The dark and gray colors separately represent UAS-jhamt and Aug21-gal4 as controls. The blue color represents jhamt over-expression.(E) Daytime sleep amount of flies over-expressing jhamt showed a significant decrease in the female but an increase in the male. Data represent mean±SEM (n = 32–96). **P<0.01, ***P<0.001 determined by Student’s t test. (F) Daytime sleep-bout duration of flies over-expressing jhamt showed a significant decrease in the female but an increase in the male. Data represent mean±SEM (n = 32–96). *P<0.05, **P<0.01, ***P<0.001 determined by Student’s t test. (G) Sleep trace graph of flies with the treatments of different concentrations of JH analog. The dark, blue, green and red lines represent 0mM, 0.01mM, 0.1mM and 1mM of JH analog, respectively. (H) Daytime sleep amount after administration of 0mM, 0.01mM, 0.1mM and 1mM JH analog to adult flies. Data represent mean±SEM (n = 32–96). *P<0.05, ***P<0.001 determined by Student’s t test. (I) Daytime sleep-bout duration after administration of 0mM, 0.01mM, 0.1mM and 1mM JH analog to adult flies. Data represent mean±SEM (n = 32–96). *P<0.05, ***P<0.001 determined by Student’s t test.

https://doi.org/10.1371/journal.pgen.1007318.g001

For a loss-of-function analysis, we used a jhamt mutant (jhamt², in which JHAMT is effectively deleted [29]), which was further confirmed by immunofluorescence with anti-DmJHAMT antibody, to assess sleep regulation (Fig 2A). With loss of jhamt function, the daytime sleep amount in females significantly increased (Fig 2B and 2C). Although sleep amount of males during daytime showed no significant difference compared to control (Fig 2B and 2C), sleep quality was significantly affected by decreases of the sleep-bout duration (Fig 2D), indicating reduction of sleep quality in the mutant males. The daytime sleep amount and sleep-bout duration in the jhamt-deleted mutants of both males and females was completely or partially rescued by exposing them to 0.1mM JHA, in which sleep quality of males increased and of females decreased (Fig 2B–2D).

Download:

Fig 2.

Effects of jhamt mutation on sleep behaviors. (A) JHAMT protein could not be detected in jhamt² corpora allata cells. The blue represents DAPI, the red represents anti-JHAMT, and the third column represents a co-location of DAPI and anti-JHAMT. (B) Sleep trace graph of jhamt² and JH analog-fed jhamt² flies. The dark, blue and gray lines represent w¹¹¹⁸, jhamt² and 0.1mM JH analog-fed jhamt² flies, respectively. (C) Daytime sleep amount of jhamt² and JH analog-fed jhamt² flies. With loss of jhamt function, daytime sleep amount in the female significantly increased, and the males had a slight but not significant decrease in daytime sleep amount. The effects of administration of JH analog were consistent with Fig 1D. Data represent mean±SEM (n = 32–96). n.s. represents no significant difference, ***P< 0.001 determined by Student’s t test. (D) Daytime sleep-bout duration of jhamt² and JH analog-fed jhamt² flies. The results were generally consistent with their daytime sleep, except that the sleep-bout duration of the male exhibited a significant difference between w¹¹¹⁸ and jhamt². Data represent mean±SEM (n = 32–96). *P<0.05, **P<0.01, ***P< 0.001 determined by Student’s t test.

https://doi.org/10.1371/journal.pgen.1007318.g002

All these results above indicate that males and females exhibit a JH-responsive sexually dimorphic sleep phenotype—a long sleep phenotype in males and a short sleep phenotype in females. An increase of JH increases the sexually dimorphic sleep phenotype, and a decrease of JH decreases this difference and blurs the gender boundaries. In other words, lack of JH will result in feminization of male sleep and masculinization of female sleep.

Effects of JH receptors on sleep

The Drosophila germ cell-expressed (GCE) and methoprene-tolerant (MET), both belonging to bHLH-PAS transcription factors, are products of two paralogous genes on the X chromosome. Both proteins potentially mediate the effects of JH as candidate JH receptors[30]. In this study, gce^2.5k and met²⁷ (both of them are null alleles) were used to further investigate the function of the JH signaling pathway on daytime sleep. Results showed that the gce deletion mutant also exhibited sexually dimorphic effects on sleep. Gce mutant male flies exhibited a short-sleep phenotype, while female flies exhibited a long-sleep phenotype. Moreover, administration of 0.1mM JHA did not rescue the sleep phenotypes, and even increasing JHA concentration to 1mM did not rescue the sleep phenotype (Fig 3A and 3B). However, in the met mutant, the males and females did not exhibit sexually dimorphic sleep phenotypes, but administration of 1mM JHA did effectively induce sexual dimorphism again (S1A and S1B Fig), indicating the met mutation may be partially involved in a JH-responsive sexually dimorphic sleep regulatory pathway, and it may be also involved in other signaling to affect flies’ daytime sleep. The sleep behavior of Gce^2.5k but not of met²⁷ phenocopied jhamt², suggesting that GCE is the main receptor to regulate Drosophila sexually dimorphic sleep in the JH signaling pathway.

Download:

Fig 3.

Effects of JH receptor gce mutations on sleep behavior. (A) Gce mutation increased the female daytime sleep amount and decreased the male daytime sleep amount, while neither 0.1mM nor 1mM JH analog could rescue the mutant phenotypes. Data represent mean±SEM (n = 32–96). n.s. represents no significant difference, **P<0.01, ***P< 0.001 determined by Student’s t test. (B) Sleep-bout duration of gce^2.5k flies was consistent with their daytime sleep amount. Data represent mean±SEM (n = 32–96). n.s. represents no significant difference, **P<0.01, ***P< 0.001 determined by Student’s t test.

https://doi.org/10.1371/journal.pgen.1007318.g003

JH-GCE signaling regulates expressions of sex determination genes

To determine how the JH pathway acts on sexual sleep differentiation, we focused on the JH-GCE signaling pathway effects on the sex determination. Sex-lethal (sxl), a key upstream gene of sex-determination in female flies, is well known to establish and maintain sex determination in females[31], while Fru^M, a male-specific upstream gene for sex determination, has been shown to contribute to sexual dimorphism by sculpting the sexually dimorphic nervous system[32]. In this study, we compared mRNA levels from both female sxl and tra and male fru^M genes among w¹¹¹⁸ (as control), jhamt² and gce^2.5k, and found their transcript levels in the jhamt and gce mutants almost decreased to half compared with control levels (Fig 4A, 4B & 4E). Because SXL is mainly expressed in female germline cells[33], we determined SXL protein expression in female ovaries of wild-type w¹¹¹⁸, jhamt² and gce^2.5k. Results showed that SXL protein expression was much less in both jhamt and gce deletion mutants than in w¹¹¹⁸ controls (Fig 4C). Abnormal expression of SXL in female flies suggested that the female sex differentiation pathway, mediated by SXL and TRA, may be affected in jhamt and gce mutants. We further used elav-gal4 to drive gce dsRNA expression in a large field of central neurons and the results showed that the sleep pattern of female flies was much closer to that of males (Fig 4D). FRU^M protein is broadly expressed in the central nervous system of males, and the gce mutation led to reduced FRU^M protein in the PI neurons (Fig 4F), which have been reported to be involved in locomotor feminization[16]. Similarly, the density of FRU^M also decreased to a certain degree in jhamt² males (Fig 4F). Furthermore, we specifically used a fru driver (fru(16)-gal4) to drive gce dsRNA expression and found the male flies’ daytime sleep significantly reduced as assessed by both the daytime sleep amount and the daytime sleep-bout duration (Fig 4G). These results indicate that JH-GCE pathway is important for regulating sexual determination genes and sleep behaviors via related neurons.

Download:

Fig 4. JH-GCE signaling pathway affected sex-related gene expression.

(A, B and E) Sxl, tra1 and fru relative transcription levels in w¹¹¹⁸, jhamt² and gce^2.5k female or male flies. Data represent mean±SEM. *P<0.05, **P<0.01, ***P< 0.001 determined by Student’s t test. (C) The protein expression of SXL in female ovary. First row: w¹¹¹⁸; Second row: jhamt²; Third row: gce^2.5k. The blue represents DAPI, the red represents anti-SXL, and the third column represents co-location of DAPI and anti-SXL. (D) Sleep analysis of female flies with gce dsRNA interference driven by elav-gal4. The color of dark, gray and blue represent UAS-gce-RNAi, elav-gal4 (as controls) and elav>gce-RNAi, respectively. The line diagram is sleep trace graph; the two histograms are daytime sleep amount and daytime sleep-bout duration quantification. The bottom left figure shows the neurons with gce dsRNA interference, as detected with a GFP reporter. (F) FRU^M protein expression in pars intercerebralis of w¹¹¹⁸, jhamt² and gce^2.5k. (G) Sleep status of male flies with gce dsRNA interference in fru(16)-gal4 neurons. The color of dark, gray and blue represent UAS-gce-RNAi, fru(16)-gal4 and fru(16)> gce-RNAi, respectively. The line diagram is sleep trace graph; the two histograms are quantified analysis of the sleep amount and sleep bout duration during daytime. The bottom left figure shows the neurons with gce dsRNA interference, as detected with a GFP reporter.

https://doi.org/10.1371/journal.pgen.1007318.g004

To further identity the function of sxl, tra and fru genes in sleep regulation, we separately used deletion or insertion mutants blocking expression of sxl, tra and fru to observe effects on sleep behavior. Trans-heterozygous mutant sxl[f2]/sxl[f18] females are viable and fertile, but their sleep pattern is similar to the male pattern with an increased sleep during daytime (Fig 5A), and heterozygous tra mutants (tra[1]/+) also exhibit increased sleep in females (Fig 5B). In contrast, a fru insertion mutation in males caused significant decreases of sleep during the daytime (Fig 5C).

Download:

Fig 5. Flies with sex differentiation-related gene mutations phenocopied jhamt² and gce^2.5k sleep behaviors.

(A and B) The top graph is sleep trace; the bottom graph is quantified analysis of daytime sleep amount and sleep-bout duration. The color of black, blue, gray and green represent yw, sxl[f2]/sxl[f18], w¹¹¹⁸ and tra[1]/+ female flies, respectively. Data represent mean±SEM (n = 64–96). **P<0.01, ***P< 0.001 determined by Student’s t test. (C) Sleep trace, daytime sleep amounts and sleep-bout durations of w¹¹¹⁸ and fru[MI15628] males. Data represent mean±SEM (n = 64–96). **P<0.01 determined by Student’s t test. (D) Sleep analysis of male flies with over-expression of traF driven by fru(16)-gal4 and Aug21-gal4. The colors of black, blue and green lines represent UAS-traF/+, fru(16) >traF and Aug21 >traF sleep traces, respectively. The bottom graph is daytime sleep amount and sleep-bout duration. Data represent mean±SEM (n = 16–32). **P<0.01, ***P< 0.001 determined by Student’s t test. (E) The top picture shows sleep traces of w¹¹¹⁸ males, Fru^Δtrafemales, Fru-F males and w¹¹¹⁸ females colored by black, blue, red and green, respectively. The bottom is quantified assays of daytime sleep amount and sleep-bout duration. Data represent mean±SEM (n = 32–64). **P<0.01, ***P< 0.001 determined by Student’s t test. n.s. represents no significance. (F) Sleep status of w¹¹¹⁸ and dsx[48]/+ colored by gray and red, respectively. Data represent mean±SEM (n = 32–64).**P<0.01determined by Student’s t test. n.s. represents no significance.

https://doi.org/10.1371/journal.pgen.1007318.g005

To verify the effects of masculinization or feminization, we simultaneously used different drivers (Aug21-gal4 and Fru(16)-gal4) to drive the expression of traF (Female-specific protein TRA) in males. Results revealed that male fly’s daytime sleep pattern is closest to the female fly’s daytime sleep when the traF of females is expressed in fru neurons of males, which is consistent with the importance of FRU-expressing neurons for the male sleep pattern. Expression of traF in the JH synthesis tissue corpus allata (with Aug21-Gal4) also significantly decreased daytime sleep amount and sleep bout duration of the male flies (Fig 5D). As for more genetic methods, Fru^Δtra, with a modified S exon with tra binding sites mutation, produces female flies exhibiting a male sleep pattern (Fig 5E), and Fru-F, with a deletion of the S exon of the fru male specific splice form, produces male flies exhibiting a female sleep pattern (Fig 5E). Taken together, these results indicate that JH-GCE signaling regulating sleep sexual dimorphism is via the sexual determination genes sxl-tra in females and fru in males, respectively, and these genes seem to be the “switch” genes in females and males to separately control the sexually dimorphic sleep pattern in D. melanogaster.

Then, we explored another sex differentiation downstream gene by using a dsx heterozygous mutant. Previous research demonstrated this gene is bifunctional, since male-specific DSX represses female sexual differentiation and vice versa for female-specific DSX[34]. Our results showed that mutation of dsx in both males and females results in an intersexual sleep pattern. Total daytime sleep decreased about 22.5% (P < 0.01) in dsx[48]/+males and increased about 34.0% in females (P < 0.01)(Fig 5F), respectively. These results indicate that dsx is important to maintain sleep sexual differentiation. Thus, all sleep analysis in the context of sex differentiation-related genes suggests that JH signaling regulates sleep sexual dimorphism through sex differentiation-related genes.

The JH-GCE signaling pathway is not related to sleep homeostasis and the circadian clock

Sleep is usually under control of homeostasis, which responds to sleep need and adjusts sleep intensity, and of the sleep time keeper, or the circadian clock[35,36]. Prolonged wakefulness increases sleep pressure which results in a rebound in the next sleep stage[37], and vice versa. To examine whether the homeostatic system is destroyed by a blocked JH signaling pathway, sleep deprivation was performed during the nighttime and sleep parameters of the next daytime were analyzed using the JH receptor mutant gce^2.5k and wild-type w¹¹¹⁸ control flies. Results showed that a functional sleep homeostatic system was completely preserved in the gce mutant (Fig 6A and 6B). We further examined the circadian parameters of jhamt and gce mutants (Fig 6C) and found that the jhamt mutation dampened the flies’ rhythm amplitude and rhythmicity. Furthermore, the gce^2.5k mutant still exhibited a robust activity rhythm and ~24h periodicity. Based on the circadian phenotypes of jhamt² and gce^2.5k, we reasoned that the JH-GCE signaling effects on sleep are circadian rhythm independent.

Download:

Fig 6.

Sleep homeostatic system was completely preserved in gce^2.5k flies. (A and B) The w¹¹¹⁸ and gce^2.5k flies exhibit sleep rebound after sleep deprivation. The black line represents sleep baseline. The red line (ZT 12 -ZT 24) represents sleep deprivation. The green line (ZT0-ZT12) represents sleep rebound. (C) Circadian phenotypes of w¹¹¹⁸, jhamt²and gce^2.5k. Data represent mean±SEM (n = 32–96).

https://doi.org/10.1371/journal.pgen.1007318.g006

Discussion

Contributions of juvenile hormone, insulin, ecdysone and sex peptide signaling to sexually dimorphic sleep

The insulin system has been reported to be involved in regulation of sleep and locomotor activity [38,39]. A previous study showed that HMGCR, an enzyme responsible for the biosynthesis of JH, is controlled by the insulin receptor (lnR)[40]. A JH inhibitor fluvastain fed to male flies led to male locomotor activity feminization, and this effect was reversed by application of a JH analog[15]. Ablation of insulin-producing neurons in the adult PI or an insulin receptor mutant were also found to abolish sexual dimorphism in locomotor activity, suggesting that the insulin signaling pathway may work in conjunction with the JH pathway to control sexually dimorphic behaviors[38]. Ecdysone, another major endocrine hormone, has been shown to regulate adult flies’ daytime sleep[41]. However, the effects of ecdysone on sleep did not involve sex differences, although there is a vital relationship between JH and ecdysone for controlling molting and metamorphosis. As a male specific product, sex peptide inhibits female flies’ siesta sleep through copulation behavior, contributing to sexually dimorphic sleep[6]. Recently another paper reported that the male’s dorsal clock neurons are more active than those in females, in correlation with the increased sleep behavior of male flies[7]. In this study, we found that wild-type males synthesized more JHAMT protein in CA cells than females, and notably old flies (30^th day adults) seem to synthesize more JHAMT than young adults (3^rd day adults), although it is a small increase in females (Fig 7A and 7B). According to our sleep behavior results, increased JHAMT should promote male sleep and inhibit female sleep. Thus, the results imply that JH may act against sleep decrease in the male aging process. Interestingly, we did find that old male flies exhibit less sleep loss than old females, whose sleep loss reaches almost 50% (Fig 7C and 7D). These phenotypes possibly indicate that aging-induced sleep loss may include effects of JH.

Download:

Fig 7.

Contributions of JH to sleep decrease in flies’ aging process. (A) JHAMT expression in 3^rd day adults and 30^th day adults. The first row: 3^rd day female; second row: 30^th day female; third row: 3^rd day male; fourth row: 30^th day male. The blue represents DAPI, the red represents anti-JHAMT, and the third column represents co-localization of DAPI and anti-JHAMT. (B) Quantification of Fig 7A. The gray column represents 3^rd day flies, the blue column represents 30^th day flies. Data represent mean±SEM (n = 5–10). (C and D) Sleep changes in both sexes in 3^rd day adults and 30^th day adults which are colored with gray and blue. Data represent mean±SEM (n = 32–64).

https://doi.org/10.1371/journal.pgen.1007318.g007

Role of JH receptors in sleep regulation

JH is an important endocrine hormone, which regulates many physiological and developmental processes through JH receptors. As the JH receptors, MET or/and GCE independently act in different functions[42]. For example, JH regulates the production of female sex pheromones via MET rather than its paralog GCE[43]. However, JH regulates nuclear receptor gene E75A in S2 cells through GCE but not MET[44]. According to the sleep behavioral assays of gce and met mutants, there is no doubt that GCE is the principal receptor for regulation of sexual dimorphism in sleep. Alternatively, met mutation caused sleep length to be significantly longer during the daytime in both sexes, suggesting that met may be involved in other regulatory pathways. A previous study reported that a MET/CYC heterodimer activates circadian rhythm-dependent gene expression in female mosquitoes[45], suggesting that met may control the circadian rhythm independent of sleep.

Sex determination genes work in collaboration with the JH signal pathway in sleep regulation

From humans to insects, morphological and behavioral differences between male and female are ubiquitous. Mating, courtship-related sensory responses and aggression have been demonstrated to be under the control of the sex determination hierarchy[46,47,48]. Tra initiated by sxl modulates or even drives various female characteristics. A recent paper has reported that myc and tra can promote sexual size dimorphism in fruit flies[49]. In fact, the idea that the sex determination system participates in the regulation of sex-specific size in stag beetles was proposed several years ago, and JH was also shown to play a crucial role[10]. With regard to behavior, TRA can feminize male locomotor behavior in transgenic flies, as described above (Ref. 15). Fru, a terminal gene of the sex determination hierarchy, orchestrates male-specific neuronal morphology and behavior[50]. Activation of male-specific dsx/fru positive P1 neurons in female brain induces male-like courtship behavior[51]. Another terminal gene (dsx) coupled with fru to regulate male sexual behavior[52]. All these sex determination genes orchestrate the characteristics of “maleness” and “femaleness”. In this study, we showed that sexually dimorphic sleep in Drosophila regulated by JH-GCE signaling is produced by the pathway of sex-specific regulatory genes differing between males and females.

A model of sexually dimorphic sleep regulated by the JH signal pathway in Drosophila

We propose a model based on the available evidence to explain this complicated sleep sexual dimorphism phenomenon (Fig 8).The pars intercerebralis sends related signals to the corpora allata. The latter initiates jhamt transcription and activates JH biosynthesis from FA. JH is released and binds to the GCE protein to promote female-specific gene transcription in female D. melanogaster. On the other hand, JH signaling acts on fru, which encodes male specific protein FRU^M in the CNS, to maintain the male sleep pattern in the absence of SXL and TRA. DSX may be also responsible for sustaining male and female sleep behaviors. Therefore, the JH signaling pathway plays a necessary role in regulation of sexual dimorphism of sleep in D. melanogaster.

Download:

Fig 8. Model of JH-GCE signals regulating sexually dimorphic sleepJH signals from Drosophila brain (PI possibly) target sex differentiation-related genes and sustain male- and female-sleep types.

The black arrow represents JH signals, while the red and green arrows represent JH/GCE-FRU and JH/GCE-SXL-TRA pathways, respectively.

https://doi.org/10.1371/journal.pgen.1007318.g008

Materials and methods

Fly strains

The following strains were used in this study: UAS-jhamt, UAS-gce-RNAi, Aug21-gal4, jhamt-gal4, jhamt², gce^2.5k and met²⁷, which were previously reported (Ref. 26, Ref. 29 and Ref. 30). UAS-traF, fru(16)-gal4, Fru^Δtra and Fru-F were obtained from Yi Rao, Yufeng Pan and Chuan Zhou’s labs, respectively. Dsx[48]/TM2, tra[1]/TM2, Sxl[f2]/FM7c and Sxl[f18]/FM7c were purchased from the Bloomington Drosophila Stock Center. Flies were reared at 25°C and 65% relative humidity on a standard cornmeal-yeast-agar medium in a 12h light/12h dark cycle.

Sleep behavioral assays

Three- to five-day-old flies were housed in monitor tubes (5[W] × 65[L] mm) with fly food. Experiments were performed in an incubator at a temperature of 25 ± 1°C and a relative humidity of 65%. Light was turned on at ZT0 (local time 06:30) and off at ZT12 (local time 18:30). The sleep activity was recorded using the Drosophila Activity Monitoring System (Trikinetics, Waltham, MA). The details for the experimental protocol and data analysis were described by Chen et al (2013).

Sleep was deprived by mechanical vibration of three seconds within each minute interval for the whole night (12h). All the sleep deprived flies were immediately fixed in liquid nitrogen for qRT-PCR assays.

JHA treatment

The juvenile hormone analog (JHA) pyriproxyfen (gift from Xiru Liang, China Agriculture University) was dissolved in 95% ethanol, and JHA-containing food was prepared by adding JHA solution to standard Drosophila food at 50–55°C to the indicated concentrations. Newly eclosed flies were placed in glass tubes with standard medium for three days and transferred to detecting tubes with food containing JHA for behavior monitoring.

Immunofluorescence

Three-day-old flies were fixed in 4% paraformaldehyde for 2 h. Then 6 to 8 flies were dissected in phosphate-buffered saline (PBS). They were washed 3 times with PBST (0.5% Triton X-100 in PBS) at room temperature. The tissue was soaked in blocking solution PNT (1% goat serum in PBS) for 1 h at room temperature and incubated with primary antibody for 16–24 h at 4°C. After 3 washes, the tissue was incubated in secondary antibody for 2 h at room temperature. The samples were analyzed with Nikon Eclipse TE2000-E and Nikon D-Eclipse (Nikon, Japan) confocal microscopes. Anti-DmJHAMT antibody was provided by Dr. Niwa. Anti-FruMale antibody was provided by the Yamamoto lab. Anti-SXL antibody was provided by DSHB at the University of Iowa.

RNA Isolation, cDNA synthesis and Quantitative real-time PCR

RNA was isolated from whole bodies of 3-day-old male and female flies using Trizol Reagent (TIANGEN, Beijing) according to the manufacturer’s protocol. RNA was reverse transcribed using Fast cDNA Reverse Transcription Kit (TIANGEN, Beijing). The Quantitative real-time PCR assay was performed using Applied Biosystem Step One Real Time PCR system (Applied Biosystem, Foster, CA, USA), RealMasterMix kit and SuperReal PreMix Plus kit (TIANGEN, Beijing). The sequences of primers are shown in Supplementary S1 Table.

Statistics analysis

Statistical analysis was performed with SPSS statistics 17.0. P values was obtained with t-test and considered significant at P < 0.05 and extremely significant at P < 0.001.

Supporting information

S1 Fig. Sleep behaviors of met²⁷ flies.

(A) Met mutation led to increases in fly daytime sleep amount in both females and males. The third column represents met²⁷ flies fed with 1mM JH analog. Data represent mean±SEM (n = 96). n.s. represents no significant difference. **P<0.01, ***P<0.001 determined by Student’s t test.

(B) Sleep-bout duration of met²⁷ flies was consistent with their daytime sleep amount. And administration of 1mM JH analog induced a sleep sexual dimorphism. Data represent mean±SEM (n = 96). n.s. represents no significant difference. *P<0.05, **P<0.01, ***P<0.001 determined by Student’s t test.

(C) Circadian phenotypes of met²⁷. Data represent mean±SEM (n = 32).

https://doi.org/10.1371/journal.pgen.1007318.s001

(TIF)

S1 Table. qRT-PCR primers used in this study.

https://doi.org/10.1371/journal.pgen.1007318.s002

(DOCX)

References

1. Ujma PP, Konrad BN, Genzel L, Bleifuss A, Simor P, et al.Sleep spindles and intelligence: evidence for a sexual dimorphism. J Neurosci. 2014; 34(49): 16358–16368. https://doi.org/10.1523/JNEUROSCI.1857-14.2014 pmid:25471574
- View Article
- PubMed/NCBI
- Google Scholar
2. Hajali V, Sheibani V, Esmaeili-Mahani S, Shabani M. Female rats are more susceptible to the deleterious effects of paradoxical sleep deprivation on cognitive performance. Behav Brain Res. 2012; 228(2): 311–318. https://doi.org/10.1016/j.bbr.2011.12.008 pmid:22192378
- View Article
- PubMed/NCBI
- Google Scholar
3. Shaw PJ, Cirelli C, Greenspan RJ, Tononi G. Correlates of sleep and waking in Drosophila melanogaster. Science. 2000; 287(5459): 1834–1837. https://doi.org/10.1126/science.287.5459.1834 pmid:10710313
- View Article
- PubMed/NCBI
- Google Scholar
4. Cirelli C, Bushey D. Sleep and wakefulness in Drosophila melanogaster. Ann N Y Acad Sci. 2008; 1129:323–329. https://doi.org/10.1196/annals.1417.017 pmid:18591491
- View Article
- PubMed/NCBI
- Google Scholar
5. Huber R, Hill SL, Holladay C, Biesiadecki M, Tononi G, et al. Sleep homeostasis in Drosophila melanogaster. Sleep. 2004; 27(4): 628–639. pmid:15282997
- View Article
- PubMed/NCBI
- Google Scholar
6. Isaac RE, Li C, Leedale AE, Shirras AD. Drosophila male sex peptide inhibits siesta sleep and promotes locomotor activity in the post-mated female. P Roy Soc B-Biol Sci. 2010; 277(1678): 65–70. https://doi.org/10.1098/rspb.2009.1236 pmid:19793753
- View Article
- PubMed/NCBI
- Google Scholar
7. Guo F, Yu J, Jung HJ, Abruzzi KC, Luo W, et al. Circadian neuron feedback controls the Drosophila sleep–activity profile. Nature. 2016; 536(7616): 292–297. https://doi.org/10.1038/nature19097 pmid:27479324
- View Article
- PubMed/NCBI
- Google Scholar
8. Khericha M, Kolenchery JB, Tauber E. Neural and non-neural contributions to sexual dimorphism of mid-day sleep inDrosophila melanogaster: a pilot study. Physiol Entomol. 2016; 41(4): 327–334. https://doi.org/10.1111/phen.12134 pmid:27840547
- View Article
- PubMed/NCBI
- Google Scholar
9. Gruntenko NE, Karpova EK, Burdina EV, Adonyeva NV, Andreenkova OV, et al. Probable mechanism of sexual dimorphism in insulin control ofDrosophila heat stress resistance. Physiol Entomol. 2016; 41(1): 59–66. https://doi.org/10.1111/phen.12125
- View Article
- Google Scholar
10. Gotoh H, Miyakawa H, Ishikawa A, Ishikawa Y, Sugime Y, et al. (2014) Developmental Link between Sex and Nutrition; doublesex Regulates Sex-Specific Mandible Growth via Juvenile Hormone Signaling in Stag Beetles. 1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA. pmid:24453990
- View Article
- PubMed/NCBI
- Google Scholar
11. Shinoda T, Itoyama K. Juvenile hormone acid methyltransferase: a key regulatory enzyme for insect metamorphosis. Proc Natl Acad Sci USA. 2003; 100(21): 11986–11991. https://doi.org/10.1073/pnas.2134232100 pmid:14530389
- View Article
- PubMed/NCBI
- Google Scholar
12. Li Y, Hernandez-Martinez S, Unnithan GC, Feyereisen R, Noriega FG. Activity of the corpora allata of adult female Aedes aegypti: effects of mating and feeding. Insect Biochem Mol Biol. 2003; 33(12): 1307–1315.pmid:14599502
- View Article
- PubMed/NCBI
- Google Scholar
13. Foltenyi K, Greenspan RJ, Newport JW. Activation of EGFR and ERK by rhomboid signaling regulates the consolidation and maintenance of sleep in Drosophila. Nat Neurosci. 2007; 10(9): 1160–1167. https://doi.org/10.1038/nn1957 pmid:17694052
- View Article
- PubMed/NCBI
- Google Scholar
14. de Velasco B, Erclik T, Shy D, Sclafani J, Lipshitz H, et al. Specification and development of the pars intercerebralis and pars lateralis, neuroendocrine command centers in the Drosophila brain. Dev Biol. 2007; 302(1): 309–323. https://doi.org/10.1016/j.ydbio.2006.09.035 pmid:17070515
- View Article
- PubMed/NCBI
- Google Scholar
15. Belgacem YH, Martin JR. Neuroendocrine control of a sexually dimorphic behavior by a few neurons of the pars intercerebralis in Drosophila. Proc Natl Acad Sci USA. 2002; 99(23): 15154–15158. https://doi.org/10.1073/pnas.232244199 pmid:12399547
- View Article
- PubMed/NCBI
- Google Scholar
16. Gatti S, Ferveur JF, Martin JR. Genetic identification of neurons controlling a sexually dimorphic behaviour. Curr Biol CB. 2000: 667–670.https://doi.org/10.1016/S0960-9822(00)00517-0 pmid:10837249
- View Article
- PubMed/NCBI
- Google Scholar
17. Yang SY, Baxter EM, Van Doren M. Phf7 controls male sex determination in the Drosophila germline. Dev Cell. 2012; 22(5): 1041–1051. https://doi.org/10.1016/j.devcel.2012.04.013 pmid:22595675
- View Article
- PubMed/NCBI
- Google Scholar
18. Kappes G, Deshpande G, Mulvey BB, Horabin JI, Schedl P. The Drosophila Myc gene, diminutive, is a positive regulator of the Sex-lethal establishment promoter, Sxl-Pe. Proc Natl Acad Sci USA. 2011; 108(4): 1543–1548. https://doi.org/10.1073/pnas.1017006108 pmid:21220321
- View Article
- PubMed/NCBI
- Google Scholar
19. Clough E, Jimenez E, Kim Y, Whitworth C, Neville MC, et al. Sex- and tissue-specific functions of Drosophila doublesex transcription factor target genes. Dev Cell. 2014; 31(6): 761–773. https://doi.org/10.1016/j.devcel.2014.11.021 pmid:25535918
- View Article
- PubMed/NCBI
- Google Scholar
20. Neville MC, Nojima T, Ashley E, Parker DJ, Walker J, et al. Male-specific fruitless isoforms target neurodevelopmental genes to specify a sexually dimorphic nervous system. Curr Biol. 2014; 24(3): 229–241. https://doi.org/10.1016/j.cub.2013.11.035 pmid:24440396
- View Article
- PubMed/NCBI
- Google Scholar
21. Shirangi TR, McKeown M. Sex in flies: What ‘body–mind’ dichotomy? Dev Biol. 2007; 306(1): 10–19. https://doi.org/10.1016/j.ydbio.2007.03.022 pmid:17475234
- View Article
- PubMed/NCBI
- Google Scholar
22. Gilbert LI, Granger NA, Roe RM. The juvenile hormones: historical facts and speculations on future research directions. Insect Biochem Molec. 2000; 30(8–9): 617–644. https://doi.org/10.1016/S0965-1748(00)00034-5 pmid:10876106
- View Article
- PubMed/NCBI
- Google Scholar
23. Niwa R, Niimi T, Honda N, Yoshiyama M, Itoyama K, et al. Juvenile hormone acid O-methyltransferase in Drosophila melanogaster. Insect Biochem Molec. 2008; 38(7): 714–720. https://doi.org/10.1016/j.ibmb.2008.04.003 pmid:18549957
- View Article
- PubMed/NCBI
- Google Scholar
24. Noriega FG, Ribeiro JM, Koener JF, Valenzuela JG, Hernandez-Martinez S, et al. Comparative genomics of insect juvenile hormone biosynthesis. Insect Biochem Mol Biol. 2006; 36(4): 366–374. https://doi.org/10.1016/j.ibmb.2006.01.013 pmid:16551550
- View Article
- PubMed/NCBI
- Google Scholar
25. Kinjoh T, Kaneko Y, Itoyama K, Mita K, Hiruma K, et al. Control of juvenile hormone biosynthesis in Bombyx mori: cloning of the enzymes in the mevalonate pathway and assessment of their developmental expression in the corpora allata. Insect Biochem Mol Biol. 2007; 37(8): 808–818. https://doi.org/10.1016/j.ibmb.2007.03.008 pmid:17628279
- View Article
- PubMed/NCBI
- Google Scholar
26. Liu Y, Sheng Z, Liu H, Wen D, He Q, et al. Juvenile hormone counteracts the bHLH-PAS transcription factors MET and GCE to prevent caspase-dependent programmed cell death in Drosophila. Development. 2009; 136(12): 2015–2025. https://doi.org/10.1242/dev.033712 pmid:19465595
- View Article
- PubMed/NCBI
- Google Scholar
27. Cirelli C. The genetic and molecular regulation of sleep: from fruit flies to humans. Nat Rev Neurosci. 2009; 10(8): 549–560. https://doi.org/10.1038/nrn2683 pmid:19617891
- View Article
- PubMed/NCBI
- Google Scholar
28. Gruntenko NE, Wen D, Karpova EK, Adonyeva NV, Liu Y, et al. Altered juvenile hormone metabolism, reproduction and stress response in Drosophila adults with genetic ablation of the corpus allatum cells. Insect Biochem Molec. 2010; 40(12): 891–897. https://doi.org/10.1016/j.ibmb.2010.09.001 pmid:20849954
- View Article
- PubMed/NCBI
- Google Scholar
29. Wen D, Rivera-Perez C, Abdou M, Jia Q, He Q, et al. Methyl farnesoate plays a dual role in regulating drosophila metamorphosis. Plos Genet. 2015; 11(3): e1005038. https://doi.org/10.1371/journal.pgen.1005038 pmid:25774983
- View Article
- PubMed/NCBI
- Google Scholar
30. Abdou MA, He Q, Wen D, Zyaan O, Wang J, et al. Drosophila Met and Gce are partially redundant in transducing juvenile hormone action. Insect Biochem Molec. 2011; 41(12): 938–945. https://doi.org/10.1016/j.ibmb.2011.09.003 pmid:21968404
- View Article
- PubMed/NCBI
- Google Scholar
31. Graham PL, Yanowitz JL, Penn JKM, Deshpande G, Schedl P. The translation initiation factor eif4e regulates the sex-specific expression of the master switch gene sxl in Drosophila melanogaster. Plos Genet. 2011; 7(e10021857). https://doi.org/10.1371/journal.pgen.1002185 pmid:21829374
- View Article
- PubMed/NCBI
- Google Scholar
32. Nojima T, Neville MC, Goodwin SF. Fruitless isoforms and target genes specify the sexually dimorphic nervous system underlying Drosophila reproductive behavior. Fly (Austin). 2014; 8(2): 95–100. https://doi.org/10.4161/fly.29132 pmid:25483248
- View Article
- PubMed/NCBI
- Google Scholar
33. Parrott BB, Chiang Y, Hudson A, Sarkar A, Guichet A, et al. Nucleoporin98-96 function is required for transit amplification divisions in the germ line of Drosophila melanogaster. Plos One. 2011; 6(9): e25087. https://doi.org/10.1371/journal.pone.0025087 pmid:21949861
- View Article
- PubMed/NCBI
- Google Scholar
34. Baker BS, Hoff G, Kaufman TC, Wolfner MF, Hazelrigg T. The doublesex locus of drosophila melanogaster and its flanking regions: a cytogenetic analysis. J Neurogenet. 1991; 127(1): 125–138.pmid:1901816
- View Article
- PubMed/NCBI
- Google Scholar
35. Seidner G, Robinson JE, Wu M, Worden K, Masek P, et al. Identification of neurons with a privileged role in sleep homeostasis in Drosophila melanogaster. Curr Biol. 2015; 25(22): 2928–2938. https://doi.org/10.1016/j.cub.2015.10.006 pmid:26526372
- View Article
- PubMed/NCBI
- Google Scholar
36. Allada R, Cirelli C, Sehgal A. Molecular mechanisms of sleep homeostasis in flies and mammals. Csh Perspect Biol. 2017; 9(8): a027730. https://doi.org/10.1101/cshperspect.a027730
- View Article
- Google Scholar
37. Liu S, Liu Q, Tabuchi M, Wu MN. Sleep drive is encoded by neural plastic changes in a dedicated circuit. Cell. 2016; 165(6): 1347–1360. https://doi.org/10.1016/j.cell.2016.04.013 pmid:27212237
- View Article
- PubMed/NCBI
- Google Scholar
38. Belgacem YH, Martin JR. Disruption of insulin pathways alters trehalose level and abolishes sexual dimorphism in locomotor activity in Drosophila. J Neurobiol. 2006; 66(1): 19–32. https://doi.org/10.1002/neu.20193 pmid:16187303
- View Article
- PubMed/NCBI
- Google Scholar
39. Cong X, Wang H, Liu Z, He C, An C, et al. Regulation of Sleep by Insulin-like Peptide System in Drosophila melanogaster. Sleep. 2015; 38(07): 1075–1083. https://doi.org/10.5665/sleep.4816 pmid:25581915
- View Article
- PubMed/NCBI
- Google Scholar
40. Belgacem YH, Martin J. Hmgcr in the corpus allatum controls sexual dimorphism of locomotor activity and body size via the insulin pathway in Drosophila. Plos One. 2007; 2(e1871). https://doi.org/10.1371/journal.pone.0000187 pmid:17264888
- View Article
- PubMed/NCBI
- Google Scholar
41. Ishimoto H, Kitamoto T. The steroid molting hormone ecdysone regulates sleep in adult Drosophila melanogaster. Genetics. 2010; 185(1): 269–281. https://doi.org/10.1534/genetics.110.114587 pmid:20215472
- View Article
- PubMed/NCBI
- Google Scholar
42. Baumann A, Barry J, Wang S, Fujiwara Y, Wilson TG. Paralogous genes involved in juvenile hormone action in Drosophila melanogaster. Genetics. 2010; 185(4): 1327–1336. https://doi.org/10.1534/genetics.110.116962 pmid:20498297
- View Article
- PubMed/NCBI
- Google Scholar
43. Bilen J, Atallah J, Azanchi R, Levine JD, Riddiford LM. Regulation of onset of female mating and sex pheromone production by juvenile hormone in Drosophila melanogaster. Proc Natl Acad Sci USA. 2013; 110(45): 18321–18326. https://doi.org/10.1073/pnas.1318119110 pmid:24145432
- View Article
- PubMed/NCBI
- Google Scholar
44. Dubrovsky EB, Dubrovskaya VA, Bernardo T, Otte V, DiFilippo R, et al. The Drosophila FTZ-F1 nuclear receptor mediates juvenile hormone activation of E75A gene expression through an intracellular pathway. J Biol Chem. 2011; 286(38): 33689–33700. https://doi.org/10.1074/jbc.M111.273458 pmid:21832074
- View Article
- PubMed/NCBI
- Google Scholar
45. Shin SW, Zou Z, Saha TT, Raikhel AS. bHLH-PAS heterodimer of methoprene-tolerant and Cycle mediates circadian expression of juvenile hormone-induced mosquito genes. Proc Natl Acad Sci USA. 2012; 109(41): 16576–16581. https://doi.org/10.1073/pnas.1214209109 pmid:23012454
- View Article
- PubMed/NCBI
- Google Scholar
46. Gleason JM. Mutations and natural genetic variation in the courtship song of Drosophila. Behav Genet. 2005; 35(3): 265–277. https://doi.org/10.1007/s10519-005-3219-y pmid:15864442
- View Article
- PubMed/NCBI
- Google Scholar
47. Savarit F, Sureau G, Cobb M, Ferveur JF. Genetic elimination of known pheromones reveals the fundamental chemical bases of mating and isolation in Drosophila. Proc Natl Acad Sci USA. 1999; 96(16): 9015–9020.https://doi.org/10.1073/pnas.96.16.9015 pmid:10430887
- View Article
- PubMed/NCBI
- Google Scholar
48. Koganezawa M, Kimura K, Yamamoto D. The neural circuitry that functions as a switch for courtship versus aggression in Drosophila males. Curr Biol. 2016; 26(11): 1395–1403. https://doi.org/10.1016/j.cub.2016.04.017 pmid:27185554
- View Article
- PubMed/NCBI
- Google Scholar
49. Wehr Mathews K, Cavegn M, Zwicky M. Sexual dimorphism of body size is controlled by dosage of the x-chromosomal gene myc and by the sex-determining gene tra in Drosophila. Genetics. 2017; 205(3): 1215–1228. https://doi.org/10.1534/genetics.116.192260 pmid:28064166
- View Article
- PubMed/NCBI
- Google Scholar
50. Ito H, Sato K, Kondo S, Ueda R, Yamamoto D. Fruitless represses robo1 transcription to shape male-specific neural morphology and behavior in Drosophila. Curr biol CB. 2016; 26(12): 1532–1542. https://doi.org/10.1016/j.cub.2016.04.067 pmid:27265393
- View Article
- PubMed/NCBI
- Google Scholar
51. Rezával C, Pattnaik S, Pavlou HJ, Nojima T, Brüggemeier B, et al. Activation of latent courtship circuitry in the brain of Drosophila females induces male-like behaviors. Curr Biol. 2016; 26(18): 2508–2515. https://doi.org/10.1016/j.cub.2016.07.021 pmid:27568592
- View Article
- PubMed/NCBI
- Google Scholar
52. Rideout EJ, Dornan AJ, Neville MC, Eadie S, Goodwin SF. Control of sexual differentiation and behavior by the doublesex gene in Drosophila melanogaster. Nat Neurosci. 2010; 13(4): 458–466. https://doi.org/10.1038/nn.2515 pmid:20305646
- View Article
- PubMed/NCBI
- Google Scholar

[ref1] 1. Ujma PP, Konrad BN, Genzel L, Bleifuss A, Simor P, et al.Sleep spindles and intelligence: evidence for a sexual dimorphism. J Neurosci. 2014; 34(49): 16358–16368. https://doi.org/10.1523/JNEUROSCI.1857-14.2014 pmid:25471574
View Article
PubMed/NCBI
Google Scholar

[2] View Article

[3] PubMed/NCBI

[4] Google Scholar

[ref2] 2. Hajali V, Sheibani V, Esmaeili-Mahani S, Shabani M. Female rats are more susceptible to the deleterious effects of paradoxical sleep deprivation on cognitive performance. Behav Brain Res. 2012; 228(2): 311–318. https://doi.org/10.1016/j.bbr.2011.12.008 pmid:22192378
View Article
PubMed/NCBI
Google Scholar

[6] View Article

[7] PubMed/NCBI

[8] Google Scholar

[ref3] 3. Shaw PJ, Cirelli C, Greenspan RJ, Tononi G. Correlates of sleep and waking in Drosophila melanogaster. Science. 2000; 287(5459): 1834–1837. https://doi.org/10.1126/science.287.5459.1834 pmid:10710313
View Article
PubMed/NCBI
Google Scholar

[10] View Article

[11] PubMed/NCBI

[12] Google Scholar

[ref4] 4. Cirelli C, Bushey D. Sleep and wakefulness in Drosophila melanogaster. Ann N Y Acad Sci. 2008; 1129:323–329. https://doi.org/10.1196/annals.1417.017 pmid:18591491
View Article
PubMed/NCBI
Google Scholar

[14] View Article

[15] PubMed/NCBI

[16] Google Scholar

[ref5] 5. Huber R, Hill SL, Holladay C, Biesiadecki M, Tononi G, et al. Sleep homeostasis in Drosophila melanogaster. Sleep. 2004; 27(4): 628–639. pmid:15282997
View Article
PubMed/NCBI
Google Scholar

[18] View Article

[19] PubMed/NCBI

[20] Google Scholar

[ref6] 6. Isaac RE, Li C, Leedale AE, Shirras AD. Drosophila male sex peptide inhibits siesta sleep and promotes locomotor activity in the post-mated female. P Roy Soc B-Biol Sci. 2010; 277(1678): 65–70. https://doi.org/10.1098/rspb.2009.1236 pmid:19793753
View Article
PubMed/NCBI
Google Scholar

[22] View Article

[23] PubMed/NCBI

[24] Google Scholar

[ref7] 7. Guo F, Yu J, Jung HJ, Abruzzi KC, Luo W, et al. Circadian neuron feedback controls the Drosophila sleep–activity profile. Nature. 2016; 536(7616): 292–297. https://doi.org/10.1038/nature19097 pmid:27479324
View Article
PubMed/NCBI
Google Scholar

[26] View Article

[27] PubMed/NCBI

[28] Google Scholar

[ref8] 8. Khericha M, Kolenchery JB, Tauber E. Neural and non-neural contributions to sexual dimorphism of mid-day sleep inDrosophila melanogaster: a pilot study. Physiol Entomol. 2016; 41(4): 327–334. https://doi.org/10.1111/phen.12134 pmid:27840547
View Article
PubMed/NCBI
Google Scholar

[30] View Article

[31] PubMed/NCBI

[32] Google Scholar

[ref9] 9. Gruntenko NE, Karpova EK, Burdina EV, Adonyeva NV, Andreenkova OV, et al. Probable mechanism of sexual dimorphism in insulin control ofDrosophila heat stress resistance. Physiol Entomol. 2016; 41(1): 59–66. https://doi.org/10.1111/phen.12125
View Article
Google Scholar

[34] View Article

[35] Google Scholar

[ref10] 10. Gotoh H, Miyakawa H, Ishikawa A, Ishikawa Y, Sugime Y, et al. (2014) Developmental Link between Sex and Nutrition; doublesex Regulates Sex-Specific Mandible Growth via Juvenile Hormone Signaling in Stag Beetles. 1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA. pmid:24453990
View Article
PubMed/NCBI
Google Scholar

[37] View Article

[38] PubMed/NCBI

[39] Google Scholar

[ref11] 11. Shinoda T, Itoyama K. Juvenile hormone acid methyltransferase: a key regulatory enzyme for insect metamorphosis. Proc Natl Acad Sci USA. 2003; 100(21): 11986–11991. https://doi.org/10.1073/pnas.2134232100 pmid:14530389
View Article
PubMed/NCBI
Google Scholar

[41] View Article

[42] PubMed/NCBI

[43] Google Scholar

[ref12] 12. Li Y, Hernandez-Martinez S, Unnithan GC, Feyereisen R, Noriega FG. Activity of the corpora allata of adult female Aedes aegypti: effects of mating and feeding. Insect Biochem Mol Biol. 2003; 33(12): 1307–1315.pmid:14599502
View Article
PubMed/NCBI
Google Scholar

[45] View Article

[46] PubMed/NCBI

[47] Google Scholar

[ref13] 13. Foltenyi K, Greenspan RJ, Newport JW. Activation of EGFR and ERK by rhomboid signaling regulates the consolidation and maintenance of sleep in Drosophila. Nat Neurosci. 2007; 10(9): 1160–1167. https://doi.org/10.1038/nn1957 pmid:17694052
View Article
PubMed/NCBI
Google Scholar

[49] View Article

[50] PubMed/NCBI

[51] Google Scholar

[ref14] 14. de Velasco B, Erclik T, Shy D, Sclafani J, Lipshitz H, et al. Specification and development of the pars intercerebralis and pars lateralis, neuroendocrine command centers in the Drosophila brain. Dev Biol. 2007; 302(1): 309–323. https://doi.org/10.1016/j.ydbio.2006.09.035 pmid:17070515
View Article
PubMed/NCBI
Google Scholar

[53] View Article

[54] PubMed/NCBI

[55] Google Scholar

[ref15] 15. Belgacem YH, Martin JR. Neuroendocrine control of a sexually dimorphic behavior by a few neurons of the pars intercerebralis in Drosophila. Proc Natl Acad Sci USA. 2002; 99(23): 15154–15158. https://doi.org/10.1073/pnas.232244199 pmid:12399547
View Article
PubMed/NCBI
Google Scholar

[57] View Article

[58] PubMed/NCBI

[59] Google Scholar

[ref16] 16. Gatti S, Ferveur JF, Martin JR. Genetic identification of neurons controlling a sexually dimorphic behaviour. Curr Biol CB. 2000: 667–670.https://doi.org/10.1016/S0960-9822(00)00517-0 pmid:10837249
View Article
PubMed/NCBI
Google Scholar

[61] View Article

[62] PubMed/NCBI

[63] Google Scholar

[ref17] 17. Yang SY, Baxter EM, Van Doren M. Phf7 controls male sex determination in the Drosophila germline. Dev Cell. 2012; 22(5): 1041–1051. https://doi.org/10.1016/j.devcel.2012.04.013 pmid:22595675
View Article
PubMed/NCBI
Google Scholar

[65] View Article

[66] PubMed/NCBI

[67] Google Scholar

[ref18] 18. Kappes G, Deshpande G, Mulvey BB, Horabin JI, Schedl P. The Drosophila Myc gene, diminutive, is a positive regulator of the Sex-lethal establishment promoter, Sxl-Pe. Proc Natl Acad Sci USA. 2011; 108(4): 1543–1548. https://doi.org/10.1073/pnas.1017006108 pmid:21220321
View Article
PubMed/NCBI
Google Scholar

[69] View Article

[70] PubMed/NCBI

[71] Google Scholar

[ref19] 19. Clough E, Jimenez E, Kim Y, Whitworth C, Neville MC, et al. Sex- and tissue-specific functions of Drosophila doublesex transcription factor target genes. Dev Cell. 2014; 31(6): 761–773. https://doi.org/10.1016/j.devcel.2014.11.021 pmid:25535918
View Article
PubMed/NCBI
Google Scholar

[73] View Article

[74] PubMed/NCBI

[75] Google Scholar

[ref20] 20. Neville MC, Nojima T, Ashley E, Parker DJ, Walker J, et al. Male-specific fruitless isoforms target neurodevelopmental genes to specify a sexually dimorphic nervous system. Curr Biol. 2014; 24(3): 229–241. https://doi.org/10.1016/j.cub.2013.11.035 pmid:24440396
View Article
PubMed/NCBI
Google Scholar

[77] View Article

[78] PubMed/NCBI

[79] Google Scholar

[ref21] 21. Shirangi TR, McKeown M. Sex in flies: What ‘body–mind’ dichotomy? Dev Biol. 2007; 306(1): 10–19. https://doi.org/10.1016/j.ydbio.2007.03.022 pmid:17475234
View Article
PubMed/NCBI
Google Scholar

[81] View Article

[82] PubMed/NCBI

[83] Google Scholar

[ref22] 22. Gilbert LI, Granger NA, Roe RM. The juvenile hormones: historical facts and speculations on future research directions. Insect Biochem Molec. 2000; 30(8–9): 617–644. https://doi.org/10.1016/S0965-1748(00)00034-5 pmid:10876106
View Article
PubMed/NCBI
Google Scholar

[85] View Article

[86] PubMed/NCBI

[87] Google Scholar

[ref23] 23. Niwa R, Niimi T, Honda N, Yoshiyama M, Itoyama K, et al. Juvenile hormone acid O-methyltransferase in Drosophila melanogaster. Insect Biochem Molec. 2008; 38(7): 714–720. https://doi.org/10.1016/j.ibmb.2008.04.003 pmid:18549957
View Article
PubMed/NCBI
Google Scholar

[89] View Article

[90] PubMed/NCBI

[91] Google Scholar

[ref24] 24. Noriega FG, Ribeiro JM, Koener JF, Valenzuela JG, Hernandez-Martinez S, et al. Comparative genomics of insect juvenile hormone biosynthesis. Insect Biochem Mol Biol. 2006; 36(4): 366–374. https://doi.org/10.1016/j.ibmb.2006.01.013 pmid:16551550
View Article
PubMed/NCBI
Google Scholar

[93] View Article

[94] PubMed/NCBI

[95] Google Scholar

[ref25] 25. Kinjoh T, Kaneko Y, Itoyama K, Mita K, Hiruma K, et al. Control of juvenile hormone biosynthesis in Bombyx mori: cloning of the enzymes in the mevalonate pathway and assessment of their developmental expression in the corpora allata. Insect Biochem Mol Biol. 2007; 37(8): 808–818. https://doi.org/10.1016/j.ibmb.2007.03.008 pmid:17628279
View Article
PubMed/NCBI
Google Scholar

[97] View Article

[98] PubMed/NCBI

[99] Google Scholar

[ref26] 26. Liu Y, Sheng Z, Liu H, Wen D, He Q, et al. Juvenile hormone counteracts the bHLH-PAS transcription factors MET and GCE to prevent caspase-dependent programmed cell death in Drosophila. Development. 2009; 136(12): 2015–2025. https://doi.org/10.1242/dev.033712 pmid:19465595
View Article
PubMed/NCBI
Google Scholar

[101] View Article

[102] PubMed/NCBI

[103] Google Scholar

[ref27] 27. Cirelli C. The genetic and molecular regulation of sleep: from fruit flies to humans. Nat Rev Neurosci. 2009; 10(8): 549–560. https://doi.org/10.1038/nrn2683 pmid:19617891
View Article
PubMed/NCBI
Google Scholar

[105] View Article

[106] PubMed/NCBI

[107] Google Scholar

[ref28] 28. Gruntenko NE, Wen D, Karpova EK, Adonyeva NV, Liu Y, et al. Altered juvenile hormone metabolism, reproduction and stress response in Drosophila adults with genetic ablation of the corpus allatum cells. Insect Biochem Molec. 2010; 40(12): 891–897. https://doi.org/10.1016/j.ibmb.2010.09.001 pmid:20849954
View Article
PubMed/NCBI
Google Scholar

[109] View Article

[110] PubMed/NCBI

[111] Google Scholar

[ref29] 29. Wen D, Rivera-Perez C, Abdou M, Jia Q, He Q, et al. Methyl farnesoate plays a dual role in regulating drosophila metamorphosis. Plos Genet. 2015; 11(3): e1005038. https://doi.org/10.1371/journal.pgen.1005038 pmid:25774983
View Article
PubMed/NCBI
Google Scholar

[113] View Article

[114] PubMed/NCBI

[115] Google Scholar

[ref30] 30. Abdou MA, He Q, Wen D, Zyaan O, Wang J, et al. Drosophila Met and Gce are partially redundant in transducing juvenile hormone action. Insect Biochem Molec. 2011; 41(12): 938–945. https://doi.org/10.1016/j.ibmb.2011.09.003 pmid:21968404
View Article
PubMed/NCBI
Google Scholar

[117] View Article

[118] PubMed/NCBI

[119] Google Scholar

[ref31] 31. Graham PL, Yanowitz JL, Penn JKM, Deshpande G, Schedl P. The translation initiation factor eif4e regulates the sex-specific expression of the master switch gene sxl in Drosophila melanogaster. Plos Genet. 2011; 7(e10021857). https://doi.org/10.1371/journal.pgen.1002185 pmid:21829374
View Article
PubMed/NCBI
Google Scholar

[121] View Article

[122] PubMed/NCBI

[123] Google Scholar

[ref32] 32. Nojima T, Neville MC, Goodwin SF. Fruitless isoforms and target genes specify the sexually dimorphic nervous system underlying Drosophila reproductive behavior. Fly (Austin). 2014; 8(2): 95–100. https://doi.org/10.4161/fly.29132 pmid:25483248
View Article
PubMed/NCBI
Google Scholar

[125] View Article

[126] PubMed/NCBI

[127] Google Scholar

[ref33] 33. Parrott BB, Chiang Y, Hudson A, Sarkar A, Guichet A, et al. Nucleoporin98-96 function is required for transit amplification divisions in the germ line of Drosophila melanogaster. Plos One. 2011; 6(9): e25087. https://doi.org/10.1371/journal.pone.0025087 pmid:21949861
View Article
PubMed/NCBI
Google Scholar

[129] View Article

[130] PubMed/NCBI

[131] Google Scholar

[ref34] 34. Baker BS, Hoff G, Kaufman TC, Wolfner MF, Hazelrigg T. The doublesex locus of drosophila melanogaster and its flanking regions: a cytogenetic analysis. J Neurogenet. 1991; 127(1): 125–138.pmid:1901816
View Article
PubMed/NCBI
Google Scholar

[133] View Article

[134] PubMed/NCBI

[135] Google Scholar

[ref35] 35. Seidner G, Robinson JE, Wu M, Worden K, Masek P, et al. Identification of neurons with a privileged role in sleep homeostasis in Drosophila melanogaster. Curr Biol. 2015; 25(22): 2928–2938. https://doi.org/10.1016/j.cub.2015.10.006 pmid:26526372
View Article
PubMed/NCBI
Google Scholar

[137] View Article

[138] PubMed/NCBI

[139] Google Scholar

[ref36] 36. Allada R, Cirelli C, Sehgal A. Molecular mechanisms of sleep homeostasis in flies and mammals. Csh Perspect Biol. 2017; 9(8): a027730. https://doi.org/10.1101/cshperspect.a027730
View Article
Google Scholar

[141] View Article

[142] Google Scholar

[ref37] 37. Liu S, Liu Q, Tabuchi M, Wu MN. Sleep drive is encoded by neural plastic changes in a dedicated circuit. Cell. 2016; 165(6): 1347–1360. https://doi.org/10.1016/j.cell.2016.04.013 pmid:27212237
View Article
PubMed/NCBI
Google Scholar

[144] View Article

[145] PubMed/NCBI

[146] Google Scholar

[ref38] 38. Belgacem YH, Martin JR. Disruption of insulin pathways alters trehalose level and abolishes sexual dimorphism in locomotor activity in Drosophila. J Neurobiol. 2006; 66(1): 19–32. https://doi.org/10.1002/neu.20193 pmid:16187303
View Article
PubMed/NCBI
Google Scholar

[148] View Article

[149] PubMed/NCBI

[150] Google Scholar

[ref39] 39. Cong X, Wang H, Liu Z, He C, An C, et al. Regulation of Sleep by Insulin-like Peptide System in Drosophila melanogaster. Sleep. 2015; 38(07): 1075–1083. https://doi.org/10.5665/sleep.4816 pmid:25581915
View Article
PubMed/NCBI
Google Scholar

[152] View Article

[153] PubMed/NCBI

[154] Google Scholar

[ref40] 40. Belgacem YH, Martin J. Hmgcr in the corpus allatum controls sexual dimorphism of locomotor activity and body size via the insulin pathway in Drosophila. Plos One. 2007; 2(e1871). https://doi.org/10.1371/journal.pone.0000187 pmid:17264888
View Article
PubMed/NCBI
Google Scholar

[156] View Article

[157] PubMed/NCBI

[158] Google Scholar

[ref41] 41. Ishimoto H, Kitamoto T. The steroid molting hormone ecdysone regulates sleep in adult Drosophila melanogaster. Genetics. 2010; 185(1): 269–281. https://doi.org/10.1534/genetics.110.114587 pmid:20215472
View Article
PubMed/NCBI
Google Scholar

[160] View Article

[161] PubMed/NCBI

[162] Google Scholar

[ref42] 42. Baumann A, Barry J, Wang S, Fujiwara Y, Wilson TG. Paralogous genes involved in juvenile hormone action in Drosophila melanogaster. Genetics. 2010; 185(4): 1327–1336. https://doi.org/10.1534/genetics.110.116962 pmid:20498297
View Article
PubMed/NCBI
Google Scholar

[164] View Article

[165] PubMed/NCBI

[166] Google Scholar

[ref43] 43. Bilen J, Atallah J, Azanchi R, Levine JD, Riddiford LM. Regulation of onset of female mating and sex pheromone production by juvenile hormone in Drosophila melanogaster. Proc Natl Acad Sci USA. 2013; 110(45): 18321–18326. https://doi.org/10.1073/pnas.1318119110 pmid:24145432
View Article
PubMed/NCBI
Google Scholar

[168] View Article

[169] PubMed/NCBI

[170] Google Scholar

[ref44] 44. Dubrovsky EB, Dubrovskaya VA, Bernardo T, Otte V, DiFilippo R, et al. The Drosophila FTZ-F1 nuclear receptor mediates juvenile hormone activation of E75A gene expression through an intracellular pathway. J Biol Chem. 2011; 286(38): 33689–33700. https://doi.org/10.1074/jbc.M111.273458 pmid:21832074
View Article
PubMed/NCBI
Google Scholar

[172] View Article

[173] PubMed/NCBI

[174] Google Scholar

[ref45] 45. Shin SW, Zou Z, Saha TT, Raikhel AS. bHLH-PAS heterodimer of methoprene-tolerant and Cycle mediates circadian expression of juvenile hormone-induced mosquito genes. Proc Natl Acad Sci USA. 2012; 109(41): 16576–16581. https://doi.org/10.1073/pnas.1214209109 pmid:23012454
View Article
PubMed/NCBI
Google Scholar

[176] View Article

[177] PubMed/NCBI

[178] Google Scholar

[ref46] 46. Gleason JM. Mutations and natural genetic variation in the courtship song of Drosophila. Behav Genet. 2005; 35(3): 265–277. https://doi.org/10.1007/s10519-005-3219-y pmid:15864442
View Article
PubMed/NCBI
Google Scholar

[180] View Article

[181] PubMed/NCBI

[182] Google Scholar

[ref47] 47. Savarit F, Sureau G, Cobb M, Ferveur JF. Genetic elimination of known pheromones reveals the fundamental chemical bases of mating and isolation in Drosophila. Proc Natl Acad Sci USA. 1999; 96(16): 9015–9020.https://doi.org/10.1073/pnas.96.16.9015 pmid:10430887
View Article
PubMed/NCBI
Google Scholar

[184] View Article

[185] PubMed/NCBI

[186] Google Scholar

[ref48] 48. Koganezawa M, Kimura K, Yamamoto D. The neural circuitry that functions as a switch for courtship versus aggression in Drosophila males. Curr Biol. 2016; 26(11): 1395–1403. https://doi.org/10.1016/j.cub.2016.04.017 pmid:27185554
View Article
PubMed/NCBI
Google Scholar

[188] View Article

[189] PubMed/NCBI

[190] Google Scholar

[ref49] 49. Wehr Mathews K, Cavegn M, Zwicky M. Sexual dimorphism of body size is controlled by dosage of the x-chromosomal gene myc and by the sex-determining gene tra in Drosophila. Genetics. 2017; 205(3): 1215–1228. https://doi.org/10.1534/genetics.116.192260 pmid:28064166
View Article
PubMed/NCBI
Google Scholar

[192] View Article

[193] PubMed/NCBI

[194] Google Scholar

[ref50] 50. Ito H, Sato K, Kondo S, Ueda R, Yamamoto D. Fruitless represses robo1 transcription to shape male-specific neural morphology and behavior in Drosophila. Curr biol CB. 2016; 26(12): 1532–1542. https://doi.org/10.1016/j.cub.2016.04.067 pmid:27265393
View Article
PubMed/NCBI
Google Scholar

[196] View Article

[197] PubMed/NCBI

[198] Google Scholar

[ref51] 51. Rezával C, Pattnaik S, Pavlou HJ, Nojima T, Brüggemeier B, et al. Activation of latent courtship circuitry in the brain of Drosophila females induces male-like behaviors. Curr Biol. 2016; 26(18): 2508–2515. https://doi.org/10.1016/j.cub.2016.07.021 pmid:27568592
View Article
PubMed/NCBI
Google Scholar

[200] View Article

[201] PubMed/NCBI

[202] Google Scholar

[ref52] 52. Rideout EJ, Dornan AJ, Neville MC, Eadie S, Goodwin SF. Control of sexual differentiation and behavior by the doublesex gene in Drosophila melanogaster. Nat Neurosci. 2010; 13(4): 458–466. https://doi.org/10.1038/nn.2515 pmid:20305646
View Article
PubMed/NCBI
Google Scholar

[204] View Article

[205] PubMed/NCBI

[206] Google Scholar

Figures

Abstract

Author summary

Introduction

Results

Effects of JH on sleep

Effects of JH receptors on sleep

JH-GCE signaling regulates expressions of sex determination genes

The JH-GCE signaling pathway is not related to sleep homeostasis and the circadian clock

Discussion

Contributions of juvenile hormone, insulin, ecdysone and sex peptide signaling to sexually dimorphic sleep

Role of JH receptors in sleep regulation

Sex determination genes work in collaboration with the JH signal pathway in sleep regulation

A model of sexually dimorphic sleep regulated by the JH signal pathway in Drosophila

Materials and methods

Fly strains

Sleep behavioral assays

JHA treatment

Immunofluorescence

RNA Isolation, cDNA synthesis and Quantitative real-time PCR

Statistics analysis

Supporting information

S1 Fig. Sleep behaviors of met27 flies.

S1 Table. qRT-PCR primers used in this study.

References

S1 Fig. Sleep behaviors of met²⁷ flies.