Expression Of Ecdysone Receptor Research Articles

N(alpha)-acetyltransferase 40-mediated histone acetylation plays an important role in ecdysone regulation of metamorphosis in the red flour beetle, Tribolium castaneum

Histone acetylation, a crucial epigenetic modification, is governed by histone acetyltransferases (HATs), that regulate many biological processes. Functions of HATs in insects are not well understood. We identified 27 HATs and determined their functions using RNA interference (RNAi) in the model insect, Tribolium castaneum. Among HATs studied, N-alpha-acetyltransferase 40 (NAA40) knockdown caused a severe phenotype of arrested larval development. The steroid hormone, ecdysone induced NAA40 expression through its receptor, EcR (ecdysone receptor). Interestingly, ecdysone-induced NAA40 regulates EcR expression. NAA40 acetylates histone H4 protein, associated with the promoters of ecdysone response genes: EcR, E74, E75, and HR3, and causes an increase in their expression. In the absence of ecdysone and NAA40, histone H4 methylation by arginine methyltransferase 1 (ART1) suppressed the above genes. However, elevated ecdysone levels at the end of the larval period induced NAA40, promoting histone H4 acetylation and increasing the expression of ecdysone response genes. NAA40 is also required for EcR, and steroid-receptor co-activator (SRC) mediated induction of E74, E75, and HR3. These findings highlight the key role of ecdysone-induced NAA40-mediated histone acetylation in the regulation of metamorphosis.

Effects of Light Spectrum on Survival, Growth, Physiological, and Biochemical Indices of Redclaw Crayfish (Cherax quadricarinatus) Juveniles

The spectrum is a key environmental factor, and light-emitting diodes (LEDs) can influence the growth and development of crustaceans by altering the composition of the spectrum. This study conducted a 30-day experiment to investigate the effects of five LED spectra (red, yellow, blue, green, and white light) on the growth, antioxidant and immune enzyme activities, stress hormone levels, and the expression of α-amylase (α-AMY), ecdysone receptor (EcR) and retinoid X receptor (RXR) genes in juvenile redclaw crayfish (Cherax quadricarinatus). The results show that the survival rate of juveniles is markedly higher in the yellow and red-light groups than in the other three groups (P<0.05). The green light group exhibits the lowest survival rate, yet it demonstrates the highest weight gain rate and specific growth rate. Regarding enzyme activity and hormone levels, the yellow light group shows the lowest malondialdehyde content, with higher superoxide dismutase and acid phosphatase activity than the other groups; no significant differences are observed in lysozyme activity among the groups (P>0.05). The melatonin content in the green and blue light groups is significantly higher than that in the other three groups (P<0.01). In terms of growth gene expression, the expression of α-AMY, EcR, and RXR in juvenile C. quadricarinatus is regulated by the spectrum. In conclusion, when raised under the yellow light spectrum, juvenile C. quadricarinatus displays elevated survival rates, rapid growth, and robust antioxidant and immune defenses. This study provides important technical parameters for optimizing and enhancing the industrial cultivation of juvenile C. quadricarinatus.

Effects of astaxanthin in diet on antioxidant capacity and molting-related genes expression of larvae giant freshwater prawn (Macrobrachium rosenbergii)

This experiment was carried out to determine the effects of astaxanthin on reactive oxide species (ROS), antioxidant capacity as well as molting-related genes expression in larvae of giant freshwater prawn, Macrobrachium rosenbergii. A total of 90,000 larvae were assigned to five treatments with six replicates each. Astaxanthin mixed in steamed eggs with graded effective content as 0 (control, A0), 40 (A40), 80 (A80), 160 (A160) and 320 (A320) mg kg−1 were fed to larvae for 14 days. The results showed that ROS of larvae in astaxanthin treatments was significantly lower than the control (P < 0.05). T-AOC, T-SOD and GR activities under astaxanthin treatments generally showed a downward trend and decreased significantly than the control (P < 0.05). No significant difference was observed towards CAT activities among A40 and A80, but the values were markedly greater than A0, A160, A320 (P < 0.05). Larvae in A80 exhibited stronger MDA content than the other (P < 0.05). GSH-Px activity was significantly highest in A320 than the other treatments (P < 0.05). The expression of molt-inhibiting hormone (MIH) in A40 and A80 enhanced markedly than the control (P < 0.05). The elevated expression of Ecdysone receptor (EcR) was recorded only in A160 and A320 (P < 0.05). Chitinase (CHIT) expression among treatments was significantly downregulated than the control except for A40. Heat shock protein (HSP21) and chitooligosaccharidolytic beta-N-acetylglucosaminidase-like expression increased dramatically in A160 and A320 (P < 0.05). For actin 6 gene, higher expression was happened only in A40 compared to control, others presented drop expression in very low level (P < 0.05). This study provides a theoretical basis that 320 mg/kg astaxanthin was excessive level, and might decreased partial antioxidant indexes and influenced molting genes expression of larvae M. rosenbergii.

Response of moulting genes and gut microbiome to nano-plastics and copper in juvenile horseshoe crab Tachypleus tridentatus

Nanoplastics (NPs) and heavy metals are typical marine pollutants, affecting the gut microbiota composition and molting rate of marine organisms. Currently, there is a lack of research on the toxicological effects of combined exposure to horseshoe crabs. In this study, we investigated the effects of NPs and copper on the expression of molt-related genes and gut microbiome in juvenile tri-spine horseshoe crabs Tachypleus tridentatus by exposing them to NPs (100 nm, 104 particles L−1) and/or Cu2+ (10 μgL−1) in seawater for 21 days. Compared with the control group, the relative mRNA expression of ecdysone receptor (EcR), retinoid x receptor (RXR), calmodulin-A-like isoform X1 (CaM X1), and heat shock 70 kDa protein (Hsp70) were significantly increased under the combined stress of NPs and Cu2+. There were no significant differences in the diversity and abundance indices of the gut microbial population of horseshoe crabs between the NPs and/or Cu2+ groups and the control group. According to linear discriminant analysis, Oleobacillus was the most abundant microorganism in the NPs and Cu2+ stress groups. These results indicate that exposure to either NPs stress alone or combined NPs and Cu2+ stress can promote the expression levels of juvenile molting genes. NPs exposure has a greater impact on the gut microbial community structure of juvenile horseshoe crabs compared to Cu2+ exposure. This study is helpful for predicting the growth and development of horseshoe crabs under complex environmental pollution.

20-hydroxyecdysone Upregulates Ecdysone Receptor (ECR) Gene to Promote Pupation in the Honeybee, Apis mellifera Ligustica.

A heterodimeric complex of two nuclear receptors, the ecdysone receptor (ECR) and ultraspiracle (USP), transduces 20-hydroxyecdysone (20E) signaling to modulate insect growth and development. Here, we aimed to determine the relationship between ECR and 20E during larval metamorphosis and also the specific roles of ECR during larval-adult transition in Apis mellifera. We found that ECR gene expression peaked in the 7-day-old larvae, then decreased gradually from the pupae stage. 20E slowly reduced food consumption and then induced starvation, resulting in small-sized adults. In addition, 20E induced ECR expression to regulate larval development time. Double-stranded RNAs (dsRNAs) were prepared using common dsECR as templates. After dsECR injection, larval transition to the pupal stage was delayed, and 80% of the larvae showed prolonged pupation beyond 18 h. Moreover, the mRNA levels of shd, sro, nvd, and spo, and ecdysteroid titers were significantly decreased in ECR RNAi larvae compared with those in GFP RNAi control larvae. ECR RNAi disrupted 20E signaling during larval metamorphosis. We performed rescuing experiments by injecting 20E in ECR RNAi larvae and found that the mRNA levels of ECR, USP, E75, E93, and Br-c were not restored. 20E induced apoptosis in the fat body during larval pupation, while RNAi knockdown of ECR genes reduced apoptosis. We concluded that 20E induced ECR to modulate 20E signaling to promote honeybee pupation. These results assist our understanding of the complicated molecular mechanisms of insect metamorphosis.

Flubendiamide induced genetic and cellular damages directly influence the life cycle of the oriental leaf worm, Spodoptera litura

Indiscriminate uses of insecticide greatly damage the environment as well as non-target organisms. Thus, multiple levels of bioassays can help better management of our environment. Flubendiamide is a phthalic acid diamide insecticide that ceases the function of insect muscle leading to paralysis and death. Here, we aimed to explore the effects of Flubendiamide on the life cycle of Spodoptera litura vis-a-vis the mode of action. Fourth instar larvae of the same age (120 ± 2 h) and size were fed with different concentrations (20–80 μg/mL) of Flubendiamide for 12–72 h. We performed a pharmacokinetics study, different biochemical assays, p450, Ecdysone receptor (EcR) and other genes expression analyses by Real-Time PCR and gross damages by Dye exclusion assay and histopathology. Our results demonstrate that the mean concentration of Flubendiamide after 48 h is 9.907 μg/mL and (i) altered the molting, metamorphosis, and reproduction at 80 μg/mL (24 h) (ii) increases all oxidative stress parameters (ROS/RNS, MDA, 8OHdG), decreases oxidative defense mechanisms (SOD, CAT, GST) at 80 μg/mL (48 h) and p450 in a time and concentration-dependent manner, (iii) activates CncC/Maf apoptotic pathways at 80 μg/mL concentration at 24 h while the expression declined from 48 h onwards, (iii) downregulates the EcR expression in a time and concentration-dependent manner, which might be responsible for disturbed molting, metamorphosis, and reproduction, and (iv) increase the expression of apoptotic genes (Caspase 1, −3, and − 5), in time and concentration-dependent manner causing gross morphological and histological damages. In conclusion, indiscriminate use of this insecticide can affect the ecosystem and have the capacity to cause multiple hazardous effects on experimental organisms. Thus, it warrants further investigations to improve and optimize the integrated pest management packages, including Flubendiamide for better management.

Sex-specific effects of propiconazole on the molting of the Chinese mitten crab (Eriocheir sinensis)

Given the inevitable exposure of Eriocheir sinensis (E. sinensis) to fungicides in rice-crab co-culture systems, understanding the potential effect of fungisides is important for practical application. Molting is a crucial development process of E. sinensis, which is regulated by endocrine system and genetic factors, and is susceptible to exogenous chemicals. However, the impact of fungicides application on the molting of E. sinensis have been rarely reported. In the present study, propiconazole, a widely used fungicide for rice disease management, was found to exert potential effects on the molting of E. sinensis at residual-related level in the rice-crab co-culture fields. After 14 days of short-term exposure to propiconazole, female crabs exhibited remarkably higher levels of hemolymph ecdysone than males. When the exposure was extended to 28 days, propiconazole markedly accelerated molt-inhibiting hormone expression by 3.3-fold, ecdysone receptor expression by 7.8-fold, and crustacean retinoid X receptor expression by 9.6-fold in male crabs, while it showed the opposite effect in females with suppressed gene expression. Propiconazole also induced the activity of N-acetylglucosaminidase in male crabs rather than females during the experiments. Our study suggests that propiconazole exerts sex-specific effects on the molting of E. sinensis. The impact of propiconazole application in the rice-crab co-culture systems remains more assessment to avoid affecting the growth of cultured E. sinensis.

Mono(2-ethylhexyl) phthalate modulates lipid accumulation and reproductive signaling in Daphnia magna.

Mono(2-ethylhexyl) phthalate (MEHP) is a primary metabolite of di(2-ethylhexyl) phthalate (DEHP), which is widely used in industry as a plasticizer. Both DEHP and MEHP have been identified as endocrine disruptors affecting reproduction systems in natural aquatic environments. However, the effects of MEHP exposure on aquatic invertebrates such as Daphnia magna are still poorly understood. In the present study, lipid alterations caused by MEHP in D. magna were identified by analyzing lipid accumulation and nontarget metabolomics. In addition, reproductive endpoints were investigated. MEHP exposure under any conditions upto 2mg/L was not associated with mortality of D. magna; yet, the number of lipid droplets and the adult female daphnids reproduction rates increased after 96h of exposure and 21days of exposure, respectively. MEHP also enhanced lipid metabolism, as evident from 283 potential lipid metabolites, including glycerolipids, glycerophospholipids, and sphingolipids, identified following 48h of exposure. The MEHP-treated group exhibited significantly higher ecdysone receptor (EcR) and vitellogenin 2 (Vtg2) expression levels at 6 and 24h. At 48h, EcR and Vtg2 expression levels were downregulated in the 1 and 2mg/L MEHP exposure groups. Our data reveal that the EcR pathway changes over MEHP exposure could be associated with lipid accumulation, owing to increased lipid levels and the subsequent increase in the reproduction of MEHP-exposed D. magna.

The ecdysone receptor regulates several key physiological factors in Anopheles funestus

BackgroundMalaria is a devastating disease, transmitted by female Anopheles mosquitoes infected with Plasmodium parasites. Current insecticide-based strategies exist to control the spread of malaria by targeting vectors. However, the increase in insecticide resistance in vector populations hinder the efficacy of these methods. It is, therefore, essential to develop novel vector control methods that efficiently target transmission reducing factors such as vector density and competence. A possible vector control candidate gene, the ecdysone receptor, regulates longevity, reproduction, immunity and other physiological processes in several insects, including malaria vectors. Anopheles funestus is a prominent vector in sub-Saharan Africa, however, the function of the ecdysone receptor in this mosquito has not previously been studied. This study aimed to determine if the ecdysone receptor depletion impacts An. funestus longevity, reproduction and susceptibility to Plasmodium falciparum infection.MethodsRNA interference was used to reduce ecdysone receptor expression levels in An. funestus females and investigate how the above-mentioned phenotypes are influenced. Additionally, the expression levels of the ecdysone receptor, and reproduction genes lipophorin and vitellogenin receptor as well as the immune gene, leucine rich immune molecule 9 were determined in ecdysone receptor-depleted mosquitoes using quantitative polymerase chain reaction.ResultsEcdysone receptor-depleted mosquitoes had a shorter lifespan, impaired oogenesis, were less fertile, and had reduced P. falciparum infection intensity.ConclusionsOverall, this study provides the first experimental evidence that supports ecdysone receptor as a potential target in the development of vector control measures targeting An. funestus.

Effect of aspartame on survival, morphological and molecular levels of Polyrhachis vicina Roger (Hymenoptera, Formicidae)

Aspartame is a food additive that is widely used in the food industry. Previous data have shown that aspartame is toxic to humans and animals. However, there are few reports on the effect of aspartame on social insects living in the soil. The present study was designed to evaluate aspartame toxicity for Polyrhachis vicina Roger. Five dosages of aspartame were fed to the workers and 40 mg/kg (acceptable daily intake, ADI) aspartame was chosen to feed the 4th instar larvae, pupae, females and males in P. vicina. The results showed that the effect of aspartame on the survival rate of workers was in a time and dose dependent manner. The survival rate of 4th instar larvae, pupae and males decreased at 40 mg/kg aspartame. Meanwhile, we found that aspartame toxicity can cause weight and morphological changes. These changes may be related to the abnormal gene expression of Ecdysone receptor, Tailless and Extradenticle of P. vicina (PvEcR, PvTll and PvExd) resulting from aspartame treatment. The present study confirms the effect of aspartame toxicity on P. vicina even at the ADI dosage.

The microRNA miR-14 Regulates Egg-Laying by Targeting EcR in Honeybees (Apis mellifera).

Simple SummaryHoneybees (Apis mellifera) are important pollinators and commonly used for honey production. The oviposition behavior in honeybees is complex and errors in oviposition could affect the development of the bee colony. Ecdysone receptor (EcR) and miR-14 were previously reported to play important roles in egg-laying. Moreover, EcR was predicted to be the target gene of miR-14 and may form miR-14-EcR cross-talk. By knocking down and overexpression of miR-14 and EcR in queen model, the effect of RNA expression of miR-14 and EcR on the number of eggs laid by honeybee queens was analyzed. We found that the expression of miR-14 and EcR was associated with the egg number of queens. In specific, inhibition of miR-14 expression enhanced the egg number, while overexpression of EcR enhanced the egg number in honeybee queens.Honeybees (Apis mellifera) are important pollinators and are commonly used for honey production. The oviposition behavior in honeybees is complex and errors in oviposition could affect the development of the bee colony. Recent studies reported that RNA–RNA cross-talk played a critical role in several biological processes, including reproduction. Ecdysone receptor (EcR) and miR-14 were previously reported to play important roles in egg-laying. Moreover, EcR was predicted to be the target gene of miR-14 and may form miR-14-EcR cross-talk. In this study, knocking down and overexpression of miR-14 and EcR in queen model were implemented. The effect of RNA expression of miR-14 and EcR on the number of eggs laid by honeybee queens were analyzed. Further, luciferase assay was used to confirm the target relation between miR-14 and 3′UTR of EcR. The results showed that the expression of miR-14 and EcR was associated with the number of eggs laid by queens. In specific, inhibition of miR-14 expression enhanced the number of eggs laid, while overexpression of EcR enhanced the number of eggs laid. Lastly, we determined that miR-14 directly targets the mRNA of EcR. These findings suggest that the cross-talk of miR-14-EcR plays an important role in the number of eggs laid by honeybee queens.

Effect of dietary egg yolk lecithin levels on survival, growth, lipid metabolism, and antioxidant capacity of early juvenile green mud crab Scylla paramamosain

This study investigated the effects of different dietary egg yolk lecithin levels (with 0% (EL0), 1% (EL1), 2% (EL2), 3% (EL3), and 4% (EL4) EL supplementation) on survival, growth, lipid metabolism, and antioxidant capacity of the early juvenile green mud crab (Scylla paramamosain). Total of 420 crabs (initial weight 8.14 ± 0.30 mg) were randomly assigned into five experimental groups (with 3 replicates). At the end of the eight-week culture experiment, the highest survival (92.86 ± 4.52%) was obtained in EL3 group. Significantly higher growth performance (final body weight (FBW), weight gain (WG), and specific growth rate (SGR)) was also observed in the EL3 group, and there was no significant difference from the EL2 group. The highest molting frequency (MF) and transcription level of ecdysone receptor (EcR) gene were also obtained in EL2 group suggested that EL could increase the MF of early juvenile S. paramamosain by up-regulating the mRNA expression of EcR and ultimately promote its growth. In addition, the higher body lipid composition along with increased EL levels suggested that EL could promote lipid absorption and disposition in S. paramamosain. Compared with other crabs, the crabs in EL2 group had significantly higher ∑n-3/∑n-6 ratio. The highest transcription level of fatty acid binding proteins (FABPs) gene also observed in EL2 group. Furthermore, the crabs in EL4 group had significantly lower transcription level of fatty acid synthase (FAS) gene. This result implied that feeding of EL supress the fatty acids synthesis but induce fatty acid transport genes in S. paramamosain. In the hepatopancreas, the content of thiobarbituric acid reactive substances (TBARS) decreased significantly with the dietary EL addition. Meanwhile, significantly higher superoxide dismutase (SOD) activity was observed in the EL2 and EL3 groups. This indicated that dietary EL improved the antioxidant capacity of S. paramamosain. According to the polynomial regression analysis of SGR, 2.56% dietary EL level could achieve the maximum growth of early juvenile S. paramamosain.

Effects of dietary lipid levels on survival, growth performance, and antioxidant ability of the early juvenile Scylla paramamosain

Five isonitrogenous diets (50% crude protein) with 7%, 9%, 11%, 13%, and 15% lipid levels (named as L7, L9, L11, L13, and L15, respectively) were designed to evaluate the effects of dietary lipid levels on survival, growth performance, body composition, and antioxidant ability of the early juvenile Scylla paramamosain. Each diet was fed to three replicates of 28 crabs with initial weight 42.23 ± 0.21 mg for 8 weeks. Crabs fed diet with lowest dietary lipid level (L7 group) obtained significantly lower survival than other crabs. The highest final body weight (FBW), weight gain (WG), specific growth rate (SGR), and molting frequency (MF) were observed in L11 group. Meanwhile, the highest mRNA expression of ecdysone receptor (EcR) also observed in L11 group. Besides, a positive correlation between body lipid contents and dietary lipid levels was observed in present study. But the mRNA expression of fatty acid synthetase (FAS) was relatively lower in L15 group. In the hepatopancreas, superoxide dismutase (SOD) activity significantly increased with increasing dietary lipid levels. On the contrary, the concentration of malondialdehyde (MDA) significantly decreased with elevating dietary lipid levels (except L15 group). Meanwhile, the lowest catalase (CAT) activity appeared in L7 group, but there was no statistical difference from other groups. Through the second order polynomial regression equation analysis of WG, the optimum dietary lipid level in the early juvenile S. paramamosain was 10.31%.

Neural activity mapping of bumble bee (Bombus ignitus) brains during foraging flight using immediate early genes

Honey bees and bumble bees belong to the same family (Apidae) and their workers exhibit a division of labor, but the style of division of labor differs between species. The molecular and neural bases of the species-specific social behaviors of Apidae workers have not been analyzed. Here, we focused on two immediate early genes, hormone receptor 38 (HR38) and early growth response gene-1 (Egr1), and late-upregulated ecdysone receptor (EcR), all of which are upregulated by foraging flight and expressed preferentially in the small-type Kenyon cells of the mushroom bodies (MBs) in the honey bee brain. Gene expression analyses in Bombus ignitus revealed that HR38 and Egr1, but not EcR, exhibited an immediate early response during awakening from CO2 anesthesia. Both premature mRNA for HR38 and mature mRNA for Egr1 were induced during foraging flight, and mRNAs for HR38 and Egr1 were sparsely detected inside the whole MB calyces. In contrast, EcR expression was higher in forager brains than in nurse bees and was expressed preferentially in the small-type Kenyon cells inside the MBs. Our findings suggest that Kenyon cells are active during foraging flight and that the function of late-upregulated EcR in the brain is conserved among these Apidae species.

Cross-resistance and Fitness Cost Analysis of Resistance to Thiamethoxam in Melon and Cotton Aphid (Hemiptera: Aphididae).

The melon/cotton aphid, Aphis gossypii Glover, is a notorious pest in many crops. The neonicotinoid insecticide thiamethoxam is widely used for A. gossypii control. To evaluate thiamethoxam resistance risk, a melon/cotton aphid strain with an extremely high level of resistance to thiamethoxam (>2,325.6-fold) was established after selection with thiamethoxam for 24 generations. Additionally, the cross-resistance pattern to other neonicotinoids and fitness were analyzed. The cross-resistance results showed the thiamethoxam-resistant strain had extremely high levels of cross-resistance against clothianidin (>311.7-fold) and nitenpyram (299.9-fold), high levels of cross-resistance against dinotefuran (142.3-fold) and acetamiprid (76.6-fold), and low cross-resistance against imidacloprid (9.3-fold). Compared with the life table of susceptible strain, the thiamethoxam-resistant strain had a relative fitness of 0.950, with significant decreases in oviposition days and fecundity and prolonged developmental duration. The molecular mechanism for fitness costs was studied by comparing the mRNA expression levels of juvenile hormone acid O-methyltransferase (JHAMT), juvenile hormone-binding protein (JHBP), juvenile hormone epoxide hydrolase (JHEH), ecdysone receptor (EcR), ultraspiracle protein (USP), and Vitellogenin (Vg) in the susceptible and thiamethoxam-resistant strains. Significant overexpression of JHEH and JHBP and downregulation of EcR and Vg expression were found in the thiamethoxam-resistant strain. These results indicate that A. gossypii has the potential to develop extremely high resistance to thiamethoxam after continuous exposure, with a considerable fitness cost and cross-resistance to other neonicotinoids.

Sublethal Effects of Triflumezopyrim on Biological Traits and Detoxification Enzyme Activities in the Small Brown Planthopper Laodelphax striatellus (Hemiptera: Delphacidae).

The small brown planthopper [Laodelphax striatellus (Fallén) (Hemiptera, Delphacidae)] is one of the most destructive insect pests of rice and has developed strong resistance to several kinds of chemical insecticides. Triflumezopyrim, a novel mesoionic insecticide developed by Corteva Agriscience (formerly DuPont Crop Protection), has efficient biological activity in controlling sucking insects, such as the planthopper. However, the effects of triflumezopyrim on the growth and reproduction of L. striatellus have not been reported. In this study, an F5 generation was obtained by conducting five rounds of insecticide screening on a sensitive L. striatellus strain (F0 generation). An age-stage life table procedure was used to evaluate the effects of a sublethal concentration (LC50) of triflumezopyrim on the biological parameters of L. striatellus. Compared with those of the F0 generation, the intrinsic rate of increase (r), the finite rate (λ), and the net reproductive rate (R0) of the F5 generation were significantly decreased; nevertheless, the average duration of life (T) was not significantly affected. The results of detoxification enzyme activity assays indicated that the glutathione S-transferase and cytochrome P450 monooxygenase (P450) activities in the F5 generation were significantly higher than those in the F0 generation. The contents of vitellogenin (Vg) and vitellogenin receptor (VgR) were also detected, and the results indicated that the contents of Vg and VgR in the F5 generation were significantly decreased compared to those in the F0 generation. Furthermore, we detected the relative expression of ecdysone receptor (EcR), Vg, and VgR in the F0 and F5 generations and found that the relative expression levels of Vg and VgR in the F5 generation female adults were obviously lower than those in the F0 generation (P < 0.05), whereas the relative expression of EcR was slightly increased, although the difference was not significant (P > 0.05). Based on these results, a sublethal concentration (median lethal concentration, LC50) of triflumezopyrim may inhibit the generational growth and reproduction of L. striatellus. Moreover, our results may provide a reference for further studies of the suitability and resistance mechanisms of L. striatellus subjected to a sublethal dose of triflumezopyrim.

Biochemical and molecular changes mediated by plasticizer diethyl phthalate in Chironomus circumdatus (bloodworms)

Plasticizers are used as additives in making plastics. Diethyl phthalate (DEP) is one of the majorly used plasticizers in various products. When plastic materials are dumped in an aquatic system, there is an increase in chances of DEP leaching out and getting deposited in water. Thus the current work focuses on studying the effect of DEP on the larval stages of Chironomus circumdatus. In this study it was found that there was an increase in lipid peroxidation levels indicating imposition of oxidative stress on these larvae due to the exposure of DEP. Changes in the levels of carbohydrates and lipids were also seen. To reduce these effects antioxidant defense system may get activated. Thus investigations showed an increase in enzymatic activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and acetylcholinesterases (AchE) and decrease in the activity of glutathione reductase (GR). Nonenzymatic antioxidant glutathione levels were also increased during the post-recovery exposure period. Thus this indicates that both enzymatic, as well as non-enzymatic antioxidants, play a certain role in reducing the stress mediated by DEP. Up-regulation of gene expression of heat shock protein70 (hsp70) was observed, which is one of the conserved protein produced during stress response in many dipterans. Changes in the level of expression of the ecdysone receptor (EcR) gene were also seen in DEP exposed larvae. Thus the insights give us a preliminary indication that physiological and developmental adaptations may take place in these organisms to persist in the DEP contaminated environment. CapsuleDEP mediated stress imposes changes in the metabolites and thus activation of antioxidant defense system in aquatic midges of Chironomus circumdatus. Changes in the expression of heat shock protein70 and ecdysone receptor was also seen indicating that DEP mediated stress affects at the molecular level also of the organism. These changes may help them to tolerate and live in DEP polluted water.

MiR-34 regulates reproduction by inhibiting the expression of MIH, CHH, EcR, and FAMeT genes in mud crab Scylla paramamosain.

Mud crab Scylla paramamosain is a commercially important species widely cultured in China. It is well known that the eyestalk regulates reproductive activities in crustaceans. In our previous research, we found that the miR-34 expression level in male eyestalk was significantly higher than that in females. Thus, we assumed that it may play an important role in regulating reproduction.In this study, we used bioinformatic tools to identify the target genes of miR-34 in eyestalk. Six reproduction-related genes with an intact 3'-untranslated region (UTR), including molt-inhibiting hormone (MIH), crustacean hyperglycemic hormone (CHH), vitellogenesis-inhibiting hormone, red pigment concentrating hormone, ecdysone receptor (EcR), and farnesoic acid methyltransferase (FAMeT) were identified. When the 3'-UTR plasmid vectors of the six genes were cotransfected with miR-34 mimics into 293FT cells, respectively, the luciferase activities of four genes (MIH, CHH, EcR, and FAMeT) were significantly decreased compared with that in the control group; on the contrary, when the six plasmid vectors were cotransfected with the miR-34 inhibitor respectively, the luciferase activities of four genes (MIH, CHH, EcR, and FAMeT) were significantly higher than that in the control group. When agomiR-34 and antagomiR-34 were injected into the eyestalk respectively in vivo, the expression levels of the MIH, CHH, EcR, and FAMeT genes were detected by a quantitative real-time polymerase chain reaction. The results showed that agomiR-34 suppressed the expression of the four genes, whereas antagomiR-34 enhanced their expression. These experimental results confirmed our hypothesis that miR-34 may indirectly regulate reproduction via binding to the 3'-UTRs of MIH, CHH, EcR, and FAMeT genes and suppressing their expression.

Effect of dietary cholesterol levels on growth performance, body composition and gene expression of juvenile mud crabScylla paramamosain

A 56-day feeding trial was conducted to investigate the effect of dietary cholesterol (CHOL) levels on growth performance, body composition and gene expression of juvenile mud crab (Scylla paramamosain). Four isonitrogenous and isoenergetic diets were formulated with 0.4%, 0.8%, 1.2% and 1.6% CHOL supplemented, and the final dietary CHOL concentrations were 0.72%, 1.11%, 1.49% and 1.83% respectively. Each dietary treatment was performed with three replicates (28 crabs per replicate, initial body weight 0.04 g). At the termination of the experiment, although the survival had no statistic difference in all treatments, the mud crabs fed the lowest CHOL diet had the lowest survival rate. The weight gain (WG) of mud crab significantly increased with dietary CHOL level up to 1.11% and then significantly decreased with dietary CHOL level further increased. The total-cholesterol (T-CHOL) content in whole body had an increasing trend with the dietary CHOL level increased. Besides, dietary CHOL supplement generally increased the hepatic superoxide dismutase (SOD) activity, and the mud crabs fed diets CHOL1.11 and CHOL1.49 showed significantly higher value than those fed other diets. The hepatic aspartate aminotransferase (AST) activity decreased slightly with dietary CHOL level up to 1.11% and then significantly increased with CHOL level further increased. The mRNA expression of ecdysone receptor (EcR) gene in the eyestalk obviously increased with dietary CHOL level up to 1.11% and then significantly decreased with dietary CHOL further increased. These results suggested that about 1.11% dietary CHOL seem fulfil to maintain good growth performance and healthy condition for juvenile S. paramamosain.

Dehydration triggers ecdysone-mediated recognition-protein priming and elevated anti-bacterial immune responses in Drosophila Malpighian tubule renal cells

BackgroundDrosophila is a powerful model for the study of factors modulating innate immunity. This study examines the effect of water-loss dehydration on innate immune responsiveness in the Drosophila renal system (Malpighian tubules; MTs), and how this leads to elevated host defense and contributes to immunosenescence.ResultsA short period of desiccation-elevated peptidoglycan recognition protein-LC (PGRP-LC) expression in MTs, increased antimicrobial peptide (AMP) gene induction, and protected animals from bacterial infection. We show that desiccation increased ecdysone synthesis in MTs, while inhibition of ecdysone synthesis or ecdysone receptor expression, specifically within MTs, prevented induction of PGRP-LC and reduced protection from bacterial infection. Additionally, aged flies are constitutively water-stressed and have elevated levels of ecdysone and PGRP-LC. Conversely, adults aged at high relative humidity show less water loss and have reduced expression of PGRP-LC and AMPs.ConclusionsThe Drosophila renal system is an important contributor to host defense and can modulate immune responses in an organ autonomous manner, responding to environmental changes such as desiccation. Desiccation primes immune responsiveness by elevating PGRP-LC expression specifically in MTs. In response to desiccation, ecdysone is produced in MTs and acts in a paracrine fashion to increase PGRP-LC expression, immune responsiveness, and improve host defense. This activity of the renal system may contribute to the immunosenescence observed in Drosophila.