Vg Gene Research Articles

Juvenile hormone mediates lipid storage in the oocytes of Dipetalogaster maxima

Triatomines are vectors of Chagas disease and important model organisms in insect physiology. “Kissing bugs” are obligatory hematophagous insects. A blood meal is required to successfully complete oogenesis, a process primarily controlled by juvenile hormone (JH). We used Dipetalogaster maxima as an experimental model to further understand the roles of JH in the regulation of vitellogenesis and oogenesis. A particular focus was set on the role of JH controlling lipid and protein recruitment by the oocytes. The hemolymph titer of JH III skipped bisepoxide increased after a blood meal. Following a blood meal there were increased levels of mRNAs in the fat body for the yolk protein precursors, vitellogenin (Vg) and lipophorin (Lp), as well as of their protein products in the hemolymph; mRNAs of the Vg and Lp receptors (VgR and LpR) were concomitantly up-regulated in the ovaries. Topical administration of JH induced the expression of Lp/LpR and Vg/VgR genes, and prompted the uptake of Lp and Vg in pre-vitellogenic females. Knockdown of the expression of LpR by RNA interference in fed females did not impair the Lp-mediated lipid transfer to oocytes, suggesting that the bulk of lipid acquisition by oocytes occurred by other pathways rather than by the endocytic Lp/LpR pathway. In conclusion, our results strongly suggest that JH signaling is critical for lipid storage in oocytes, by regulating Vg and Lp gene expression in the fat body as well as by modulating the expression of LpR and VgR genes in ovaries.

Expressions of conventional vitellogenin and vitellogenin-like A in worker brains are associated with a nursing task in a ponerine ant.

In eusocial insect colonies, non-reproductive workers often perform different tasks. Tasks of an individual worker are shifted depending on various factors, e.g., age and colony demography. Although a vitellogenin (Vg) gene play regulatory roles in both reproductive and non-reproductive division of labours in a honeybee, it has been shown that the insect Vg underwent multiple gene duplications and sub-functionalisation, especially in apical ant lineages. The regulatory roles of duplicated Vgs were suggested to change evolutionarily among ants, whereas such roles in phylogenetically basal ants remain unclear. Here, we examined the expression patterns of conventional Vg (CVg), Vg-like A, Vg-like B and Vg-like C, as well as Vg receptor, during the task shift in an age-dependent manner and under experimental manipulation of colony demography in a primitive ant Diacamma sp. Expressions of CVg and Vg-like A in a brain were associated with a nursing task. It is suggested that associations of brain expressions of these Vgs with worker tasks were acquired in the basal ant lineage, and that such Vg functions could have sub-functionalised in the derived ant lineage.

Lead stress affects the reproduction of Spodoptera litura but not by regulating the vitellogenin gene promoter

Lead (Pb) stress affects hormone-mediated responses (e.g., reproduction) in insects. In this study, the effects of Pb stress (12.5–50 mg Pb/kg in larval artificial diets) on the reproduction of the common cutworm Spodoptera litura (Lepidoptera: Noctuidae) were investigated after 7 generations. The results showed that Pb stress did not reduce the longevity of adult females, but 50 mg Pb/kg significantly reduced the longevity of adult males, regardless of the generation. After 50 mg Pb/kg stress for one or 7 generations, the peak time of egg-laying was delayed, and egg production and hatchability were decreased significantly. The vitellin content in eggs was significantly inhibited by Pb stress. The S. litura vitellogenin (Vg) gene promoter was cloned and analyzed. Multiple putative transcription factors were predicted for the 2321 bp Vg promoter region, including the TATA box, GATA, basic helix-loop-helix (bHLH) transcription factor, Broad-Complex (BR-C) binding sites, etc. The fragment from −2222 to −211 bp of the Vg promoter was the activation domain for Vg, whereas the region from −211 to −55 bp repressed the activity of the Vg promoter. The construct promoter (−782/+76) in Trichoplusia ni (Hi5) cells significantly improved Vg expression, which was not affected by Pb stress (1 or 10 mg/ml). Therefore, Pb stress significantly inhibited the reproduction of S. litura but not by regulating the Vg promoter.

Acetamiprid resistance and fitness costs of melon aphid, Aphis gossypii: An age-stage, two-sex life table study

The melon aphid, Aphis gossypii is a globally distributed crop pest with a wide host range. The intensive use of insecticides against this insect over several years has led to develop resistance against many insecticides including acetamiprid. Understanding the relationship between acetamiprid resistance and fitness of A. gossypii is essential to limit the spread of the resistant population in the field. In this study, age-stage, two-sex life table approach was used to investigate these relationships in the lab. Results showed that resistant strain (Ace-R) had a reduced fitness (relative fitness = 0.909) along with significantly decreased adult longevity, fecundity, net reproductive (R0), mean generation time (T) and gross reproductive rate (GRR). Compared to the susceptible strain (Ace-S), the pre-adult period and total pre-oviposition period (TPOP) were also significantly shorter in Ace-R strain. Moreover, the expression profiles of EcR, JHBP, JHAMT, JHEH, USP and Vg genes supposed to be involved in insect reproduction and development were analyzed using Quantitative Real Time PCR. The EcR, JHBP, JHAMT and USP genes were up-regulated, Vg gene was down-regulated while the mRNA level of JHEH gene was statistically same in the Ace-R strain compared to the Ace-S strain. Collectively, this study provides the occurrence and magnitude of fitness costs of A. gossypii against acetamiprid resistance and could be helpful to manage the resistance evolution in field populations.

Effect of dietary vitamin E on reproductive performance and vitellogenin gene expression in broodstock of Litopenaeus vannamei

Vitamin E (VE) is one of the most important micronutrients that influence the performance of shrimp reproduction. The effects of different dietary VE levels (0, 100, 300 and 500 mg kg-1) were investigated on the reproductive performance and vitellogenin (Vg) gene expression in Litopenaeus vannamei females which were co-fed with fresh food (two times daily) or experimental diets (two times daily) for 30 days. Sampling was carried out at the first day of the trial, prior to eyestalk ablation (ESA) (day 21) and after ESA (day 30). The reproductive parameters including hepatopancreatic index, absolute fecundity, egg diameter, latency period, and Vg gene expression were higher in the shrimp fed with VE supplemented diets than the control group (p <0.05). After ESA, the Vg gene expression significantly increased in all groups (p <0.05). As a measure of reproductive status of Pacific white shrimp, the present study evaluated correlations of Vg gene expression in hepatopancreas with the number and diameter of oocytes in gonads. Accordingly, Vg is a precursor of vitellin, which is the major yolk protein accumulated in ooplasm. These results provide evidence that dietary VE with 300 mg kg-1 may present a novel strategy and play a positive impact on reproductive performance and Vg gene expression of female shrimp.

CRISPR/Cas9-induced vitellogenin knockout lead to incomplete embryonic development in Plutella xylostella

Vitellogenin (Vg) is important for insect egg maturation and embryo development. In the present study, we characterized the molecular structure and expression profile of Vg gene, and analyzed its reproductive functions in diamondback moth, Plutella xylostella (L.), a destructive pest of cruciferous crops, using CRISPR/Cas9 system. The P. xylostella Vg (PxVg) included all conserved domains and motifs that were commonly found in most insect Vgs except for the polyserine tract. PxVg gene was highly expressed in female pupae and adults. PxVg protein was detected in eggs and female adults. PxVg was mainly expressed in the fat body and its protein was detected in most tissues, except in the midgut. CRISPR/Cas9-induced PxVg knockout successfully constructed a homozygous mutant strain with a 5-base pair nucleotide deletion. No PxVg protein was found in the mutant individuals and in their ovaries. There were no significant differences between wild (WT) and mutant (Mut-5) types of P. xylostella in terms of ovariole length and the number of fully developed oocytes in newly emerged females. No significant difference was observed in the number of eggs laid within two days, but there was a lower egg hatchability (84% for WT vs. 47% for Mut-5). This is the first study presenting the functions of Vg in ovary development, egg maturation, oviposition and embryonic development of P. xylostella. Our results suggest that the reproductive functions of Vg may be species-specific in insects. It is possible that Vg may not be the major egg yolk protein precursor in P. xylostella. Other “functional Vgs” closely involved in the yolk formation and oogenesis would need to be further explored in P. xylostella.

Vestigial suppresses apoptosis and cell migration in a manner dependent on the level of JNK-Caspase signaling in the Drosophila wing disc.

The Decapentaplegic (Dpp) and Wingless (Wg) signal pathways play important roles in numerous biological processes in Drosophila. The Drosophila vestigial (vg) gene is selectively required for wing imaginal disc cell proliferation, which is essential for the formation of the adult wing and halter structures, and is regulated by Dpp and Wg signaling. Using a Drosophila invasion model of wing epithelium, we showed herein that inhibition of Dpp or Wg signaling promoted cells to migrate across the cell lineage restrictive anterior/posterior (A/P) compartment boundary. Being downstream of both Dpp and Wg signaling, vg can block cell migration induced by loss of either pathway. In addition, suppression of vg is sufficient to induce cell migration across the A/P boundary. Transcriptomic analysis revealed potential downstream genes involved in the cell migration after suppressing vg in the wing disc. We further demonstrated that the c-Jun N-terminal kinase (JNK) signaling promoted cell migration induced by vg suppression by upregulating Caspase activity. Taken together, our results revealed the requirement of Vg for suppressing cell migration and clarified how developmental signals collaborate to stabilize cells along the compartment boundary.

Molecular Characterization of Neoseiulus barkeri Vitellogenin Genes and Vitellogenin Receptor during Reproductive Diapause.

The relationship between reproductive diapause and the genes related to vitellogenin (Vg) and its receptor (VgR) in insectoid ovarian development is still unclear. Accordingly, in the present study, we used hematoxylin and eosin staining to study the ovarian structure in the predatory mite Neoseiulus barkeri, a species that shows promise as a biological pest control agent. Staining revealed the presence of oocytes on ovary surfaces, and the oocytes were deposited as yolk granules through the intake of Vg and other nutrients with the development of the ovary. Development of the ovary stopped at the oocyte stage in diapausing adult mites, and this stage presented the same characteristics as the first day of adulthood in non-diapause female adults, where oocytes with nutrient cells, but no yolk granules are observed. In order to further explore the effects of the Vg gene and its receptor on reproduction, the sequences of the N. barkeri vitellogenin genes NbVg1, NbVg2, NbVg3, and NbVgR were analyzed using bioinformatics, and the expression levels of the NbVgs and the VgR at different developmental stages were determined by quantitative polymerase chain reaction (qPCR). The results showed that the NbVgs and NbVgR have complete domains and that the positions of many conservative regions and conservative motif are consistent. The expression levels of the NbVgs and NbVgR were highest in the ovipositional period, followed by those in the preovipositional period. The expression levels of the NbVgs and the VgR in non-diapause female adult mites were significantly higher than those in reproductive diapause female adult mites.

JNK signaling pathway regulates the development of ovaries and synthesis of vitellogenin (Vg) in the swimming crab Portunus trituberculatus.

The development of Portunus trituberculatus egg cells is directly related to the nutritional status of the fertilized egg, which affects the key production stages of offspring hatching. Vitellogenin plays a key role in the nutrient supply required for the development of the egg cells. The c-Jun N-terminal kinase (JNK) is an important member of the mitogen-activated protein kinase (MAPK) superfamily and plays an important role in cell proliferation, transformation, differentiation, and apoptosis. At present, there are no reports on the involvement of the JNK signaling pathway in the reproductive regulation of P. trituberculatus. In this study, rapid amplification of complementary DNA ends amplification technology was used to clone the full length of JNK complementary DNA, which has a length of 2094bp, including an open reading frame (ORF) of 1266bp encoding a 421-amino acid protein. The protein includes the S_TKC conserved domain with a TPY phosphorylation site, which is a typical feature of the JNK gene family. Observing tissue sections found the oocytes in the inhibitor group developed slowly, while the oocytes in the activated group showed accelerated development. Meanwhile, Portunus trituberculatus JNK and vitellogenin (Vg) genes exhibited the same trend in the hepatopancreas and ovaries, and the expression of the SP600125 group was downregulated (P < 0.05), while the anisomycin group was upregulated (P < 0.05). In addition, JNK enzyme activity and vitellin (Vn) content in the ovarian tissue showed that the JNK activity of the SP600125 group decreased, while activity increased in the anisomycin group. The accumulation of Vn content in the SP600125 group decreased, and that in the anisomycin group increased. In summary, after injection with inhibitor or activator, the JNK signaling pathway of P. trituberculatus was inhibited or activated, the accumulation of Vn in the ovary was reduced or increased, and ovarian development was inhibited or accelerated, respectively. These results indicated that the JNK signaling pathway is involved in the regulation of Vg synthesis and ovarian development in P. trituberculatus. The results of this study further add to the knowledge of the breeding biology of P. trituberculatus and provide a theoretical reference for the optimization of breeding techniques in aquaculture production systems.

&lt;p&gt;&lt;strong&gt;Effects of&lt;/strong&gt;&lt;strong&gt; &lt;/strong&gt;&lt;strong&gt;food sources on the fecundity and gene expression of vitellogenin and its receptor from &lt;/strong&gt;&lt;strong&gt;&lt;em&gt;Amblyseius eharai &lt;/em&gt;&lt;/strong&gt;&lt;strong&gt;(Acari: Phytoseiidae)&lt;/strong&gt;&lt;strong&gt;&lt;/strong&gt;&lt;/p&gt;

Vitellogenin (Vg) and Vitellogenin receptor (VgR) play key roles in the reproductive process and development of many arthropod species, including Amblyseius eharai, a dominant natural enemy of spider mites and other small pests. In this study, impacts of different food on A. eharai fecundity were investigated. Two Vg genes (AeVg1 and AeVg2) and AeVgR genes were cloned, with their expression pattern analyzed. When fed with different foods including Tetranychus cinnabarinus, Aleuroglyphus ovatus and Camellia oleifera pollen, the number of eggs for each female of A. eharai was influenced significantly (P&lt;0.05), while the hatching rate of eggs not significantly (P&gt;0.05). The open reading frames of AeVg1, AeVg2 and AeVgR were 5673, 5634 and 5597 bp, encoding 1857, 1851 and 1830 amino acids, respectively. Phylogenetic analysis of the amino acid sequences between the AeVgs and AeVgR and 35 other Vg and VgRs from other arthropod species showed that A. eharai AeVg1 was related most closely to Neoseiulus barkeri Vg1, AeVg2 was related most closely to Neoseiulus cucumeris Vg2, AeVgR was most similar to Varroa jacobsori VgR. AeVgs and AeVgR had similar expression patterns: the highest expression was observed in the oviposition female stage, and the highest expression was observed in A. eharai fed with C. oleifera pollen. Positive correlations between expression of AeVgs and AeVgR and fecundity were observed. These results will help us understand the molecular mechanism of the reproductive capacity in A. eharai.

Functional characterization of the cis-regulatory region for the vitellogenin gene in Plutella xylostella.

The vitellogenin gene promoter (VgP) is an essential cis-regulatory element that plays a significant role in transcription of the vitellogenin (Vg) gene, leading to the production of yolk protein in insects, including lepidopterans. However, the function of VgP is still not clear in Plutella xylostella. Here, we cloned a 5.1 kb DNA fragment of the cis-regulatory region adjacent to the 5' end of the Vg gene of P. xylostella (PxVg). We identified two promoter sites in that 5' upstream sequence of PxVg and performed in vitro analysis of two promoter sequences (PxVgP1, 4.9 kb, and PxVgP2, 2.9 kb) in the embryonic cell line of P. xylostella. PxVgP2 exhibited higher enhanced green fluorescent protein (EGFP) expression, so PxVgP2 was used for in vivo analysis. Strong EGFP fluorescence was observed in adult females and the fat body of females, with low expression in embryos. Our results suggest that PxVgP is an important stage-, tissue- and sex-specific endogenous cis-regulatory element in P. xylostella.

Molecular Characterization of Vitellogenin and Its Receptor in Sogatella furcifera, and Their Function in Oocyte Maturation.

The yolk protein precursor, vitellogenin (Vg), provides nutrition for embryonic development whereas the vitellogenin receptor (VgR) is responsible for the uptake of yolk protein by maturing oocytes. These two proteins are key reproduction-related proteins in insects. We cloned and characterized Vg and VgR genes in Sogatella furcifera, and investigated their function in oocyte maturation. Cloned SfVg and SfVgR have open reading frames of 6,114 and 5,796 bp, encoding 2,037 and 1,931 amino acid residues, respectively. Structural analysis indicates that SfVg has the three conserved LPD_N, DUF1943, and VWFD domains, SfVgR contains all conservative motifs of the LDLR superfamily. Both genes were highly expressed in adult females; SfVg was most highly expressed in the fat body whereas SfVgR was mainly expressed in the ovary. Knockdown of either gene reduced yolk protein deposition in oocytes and arrested oocyte maturation. However, silencing one of these two genes did not affect the transcript level of the other. These results demonstrate the role of SfVgR in transporting SfVg into oocytes. Both SfVg and SfVgR are essential for oocyte maturation in S. furcifera and both genes could potentially be targeted as means of controlling this pest.

Transcriptomics of Cherax quadricarinatus hepatopancreas during infection with Decapod iridescent virus 1 (DIV1)

Cherax quadricarinatus is a large-sized, highly fecund, and fast-growing species of freshwater crayfish, and has become one of the world's most intensely studied crustaceans. Decapod iridescent virus 1 (DIV1), a newly described species in the family Iridoviridae, is known to infect various crustaceans, including C. quadricarinatus, and may pose a new threat in the shrimp-farming industry. The present study performed de novo transcriptome sequencing of C. quadricarinatus hepatopancreas during DIV1 infection. A total of 114,784 transcripts and 56,418 genes were obtained; 1070 genes were upregulated and 775 genes were downregulated when compared with the uninfected samples (controls). Three pattern recognition receptor genes (fibrinogen-related protein, C-type lectin, and beta-1,3-glucan-binding protein) were upregulated during DIV1 infection. Among the top-30 upregulated unigenes, 9 unigenes were identified as vitellogenin (Vg) genes, and the top-3 upregulated unigenes were identified as involved in Vg lipid transport, lipid localization, and lipid transporter activity, which were all significantly over-representative GO terms in the GO enrichment analysis of total and upregulated differentially expressed genes (DEGs). Many genes associated with Jak-STAT signaling pathway, Endocytosis, Phagosome, MAPK signaling pathway, Apoptosis and Lysosome were positively modified after DIV1 infection. The predicted protein–protein interaction (PPI) analysis showed NF1 and TUBA, CRM1 and TUBB were involved in protein interactions. This research showed that DIV1 infection has a significant impact on the transcriptome profile of C. quadricarinatus hepatopancreas, and the results enhance our understanding of virus–host interactions. Furthermore, the high number of transcripts generated in the present study will provide information for identifying novel genes in the absence of a full C. quadricarinatus genome sequence.

Transcriptome analysis of the almond moth, Cadra cautella, female abdominal tissues and identification of reproduction control genes

BackgroundThe almond moth, Cadra cautella is a destructive pest of stored food commodities including dates that causes severe economic losses for the farming community worldwide. To date, no genetic information related to the molecular mechanism/strategies of its reproduction is available. Thus, transcriptome analysis of C. cautella female abdominal tissues was performed via next-generation sequencing (NGS) to recognize the genes responsible for reproduction.ResultsThe NGS was performed with an Illumina Hiseq 2000 sequencer (Beijing Genomics Institute: BGI). From the transcriptome data, 9,804,804,120 nucleotides were generated and their assemblage resulted in 62,687 unigenes. The functional annotation analyses done by different databases, annotated, 27,836 unigenes in total. The transcriptome data of C. cautella female abdominal tissue was submitted to the National Center for Biotechnology Information (accession no: PRJNA484692). The transcriptome analysis yielded several genes responsible for C. cautella reproduction including six Vg gene transcripts. Among the six Vg gene transcripts, only one was highly expressed with 3234.95 FPKM value (fragments per kilobase per million mapped reads) that was much higher than that of the other five transcripts. Higher differences in the expression level of the six Vg transcripts were confirmed by running the RT-PCR using gene specific primers, where the expression was observed only in one transcript it was named as the CcVg.ConclusionsThis is the first study to explore C. cautella reproduction control genes and it might be supportive to explore the reproduction mechanism in this pest at the molecular level. The NGS based transcriptome pool is valuable to study the functional genomics and will support to design biotech-based management strategies for C. cautella.

Vitellogenin and Vitellogenin-Like Genes in the Brown Planthopper

Vitellogenin (Vg) is precursor of vitellin. Here, we identified a Vg (NlVg) and two Vg-likes (NlVg-like1 and NlVg-like2) in the brown planthopper, Nilaparvata lugens. Phylogenetic analyses showed that NlVg-like1 and NlVg-like2 are not clustered with the conventional insect Vgs associated with vitellogenesis. Temporo-spatial expression analyses showed that the NlVg and NlVg-like2 transcript levels increased significantly 24 h after emergence and were primarily expressed in female adults. However, NlVg-like1 was expressed during all stages, and in both genders. Tissue-specific analyses showed that all three genes were most highly expressed in the fat body. The injection of double-stranded RNA targeting NlVg showed that NlVg is essential not only for oocyte development but also for nymph development. The knockdown of NlVg-like1 in female adults resulted in failure to hatch or death before eggshell emergence in 18% of offspring embryos, suggesting that NlVg-like1 plays an important role during late embryogenesis. Approximately 65% of eggs laid by females that were treated with double-stranded RNA targeting NlVg-like2 failed to hatch, indicating that NlVg-like2 plays a role in nutrition absorption during oocyte, or embryonic development. Our results illustrate the structural and functional differences among the Vg and Vg-like genes and provide potential targets for RNA-interference-based insect pest management strategies.

Identification of Halloween Genes and RNA Interference-Mediated Functional Characterization of a Halloween Gene shadow in Plutella xylostella.

Ecdysteroids play an essential role in controlling insect development and reproduction. Their pathway is regulated by a group of enzymes called Halloween gene proteins. The relationship between the Halloween genes and ecdysteroid synthesis has yet to be clearly understood in diamondback moth, Plutella xylostella (L.), a worldwide Lepidoptera pest attacking cruciferous crops and wild plants. In this study, complete sequences for six Halloween genes, neverland (nvd), shroud (sro), spook (spo), phantom (phm), disembodied (dib), shadow (sad), and shade (shd), were identified. Phylogenetic analysis revealed a strong conservation in insects, including Halloween genes of P. xylostella that was clustered with all other Lepidoptera species. Three Halloween genes, dib, sad, and shd were highly expressed in the adult stage, while nvd and spo were highly expressed in the egg and pupal stages, respectively. Five Halloween genes were highly expressed specifically in the prothorax, which is the major site of ecdysone production. However, shd was expressed predominantly in the fat body to convert ecdysone into 20-hydroxyecdysone. RNAi-based knockdown of sad, which is involved in the last step of ecdysone biosynthesis, significantly reduced the 20E titer and resulted in a longer developmental duration and lower pupation of fourth-instar larvae, as well as caused shorter ovarioles and fewer fully developed eggs of P. xylostella. Furthermore, after the knockdown of sad, the expression levels of Vg and VgR genes were significantly decreased by 77.1 and 53.0%. Meanwhile, the number of eggs laid after 3 days was significantly reduced in sad knockdown females. These results suggest that Halloween genes may play a critical role in the biosynthesis of ecdysteroids and be involved in the development and reproduction of P. xylostella. Our work provides a solid basis for understanding the functional importance of these genes, which will help to screening potential genes for pest management of P. xylostella.

Variation of protein kinase C-α expression in eyestalk removal-activated ovaries in whiteleg shrimp, Litopenaeus vannamei

The regulation of female reproduction in crustaceans is controlled by a variety of hormones. Previous studies of hormone-initiated cellular mechanisms controlling ovarian maturation focused mainly on those initiated by inhibitory hormones. In order to facilitate research on ovarian development, it is necessary to gain a better understanding of factors that promote ovarian growth on the cellular level. Here, we used eyestalk ablation to firstly induce a state of ovarian maturation in Litopenaeus vannamei. Gonadosomatic index, hemolymph vitellogenin (Vg) concentrations, and Vg gene transcript levels in the ovaries were significantly elevated at 1 and 2 weeks after eyestalk ablation (P < 0.05). Correspondingly, immunoblot analysis revealed a remarkable decrease in anti-PKC-α immunoreactivity in both cytosol and membrane fractions of ovarian tissue homogenates: it was strongly apparent in intact animals, but decreased with time after eyestalk ablation, showing a stronger tendency to do so in the membrane fraction than in the cytosol fraction. Considered overall, the data presented strongly suggest that PKC-α isoform plays a role in the regulation of ovarian growth in L. vannamei through a negative-based regulating mechanism.

Differential expression of acetylcholinesterase 1 in response to various stress factors in honey bee workers

The honey bee acetylcholinesterase 1 (AmAChE1) has been suggested to be related to stress response as judged from its elevated expression level under brood rearing-suppressed conditions. To further investigate the involvement of AmAChE1 expression in the stress response and its physiological functions, we analyzed altered expression profiles of AmAChE1 induced by diverse stress factors. In addition, transcription profiles of several heat shock protein (Hsp) genes (hsps) and the vitellogenin (Vg) gene (vg) known as general stress markers were investigated as positive references. Among the tested stress conditions, AmAChE1 expression was induced under the brood rearing-suppressed, crowding and heat shock conditions. The hsps, particularly hsp70 and hsp90, responded to seven of nine stress conditions tested, confirming that hsp expression profiles can serve as a general stress marker. Taken together, AmAChE1 expression is not suitable for using as a stress marker due to its limited response. Nevertheless, AmAChE1 expression appears to be connected, at least in part, to heat shock response and other pathways. Considering that AmAChE1 likely regulates the ACh titer particularly in non-neuronal tissues, thereby modulating the signal cascades mediated by mAChR, the AmAChE1 expression profile under different conditions likely provides important information on its physiological roles in honey bees.

Effects of low glyphosate-based herbicide concentrations on endocrine-related gene expression in the decapoda Macrobrachium potiuna.

Glyphosate-based herbicides (GBH) are the most used herbicides worldwide and are considered as endocrine-disrupting compounds (EDC) for non-target organisms. However, effects of GBH on their endocrine systems remain poorly understood. Thus, the aim of this study was to assess the effects of low concentrations of Roundup WG® on growth and reproduction process molecules in both males and females of the decapod crustacean Macrobrachium potiuna, by the relative transcript expression levels of the ecdysteroid receptor (EcR), the molt-inhibiting hormone (MIH), and the vitellogenin (Vg) genes. Prawns were exposed to three concentrations of GBH (0.0065, 0.065, and 0.28mgL-1) for 7 and 14days. The results revealed that only in males the three genes transcript levels were influenced by the GBH concentration, time of exposure, and the interaction between the concentrations and time of exposure, suggesting that males were more sensitive to GBH than females. For males, after 7days of exposure at 0.065mgL-1, EcR and MIH were over-expressed, while the Vg expression was only over-expressed after 14days. The present study highlighted that GBH impacted endocrine systems of M. potiuna. Moreover, EcR and MIH gene expressions could be promising EDC biomarkers of exposure in crustaceans. These results also indicate that GBH concentrations, considered secure by regulatory agencies, should be reviewed to minimize the effects on non-target organisms. Potential effects of glyphosate-based herbicides on the endocrine system of decapods Macrobrachium sp.

A putative direct repeat element plays a dual role in the induction and repression of insect vitellogenin-1 gene expression.

Juvenile hormones (JH) regulate wide-ranging physiological and developmental processes in insects. However, molecular mechanisms underlying JH signaling remain to be determined. Vitellogenin (Vg) is primarily an egg-yolk protein, but recently proposed to serve many functions in insects. In the female American cockroach (Periplaneta americana), vitellogenin (Vg) genes are activated by JH III and suppressed by 20-hydroxyecdysone (20E) via cis-regulatory elements in a dose-dependent manner. In the present study, the upstream promoter region (935 bp) of Vg1 was cloned to elucidate the action of these hormones. A luciferase reporter assay identified an 81 bp region in the promoter region of Vg1 (-120 to -39 bp) that we found to be critical for JH III activation and 20E suppression. This 81 bp region contains a direct repeat separated by a 2-nucleotide spacer-designated Vg1HRE- that is similar to the Drosophila ecdysone response element direct repeat 4. Moreover, nuclear proteins isolated from nymphs, males, females, and Sf9 cells successfully bound to Vg1HRE, while binding was outcompeted by a 100-fold excess of cold probe or dephosphorylated nuclear protein extracts. In addition, binding was outcompeted by other ecdysone and JH response elements with similar half-site sequences (direct repeats) but to varying extents. Ultimately, we postulate that JH III indirectly activates Vg expression by interfering with or inhibiting the phosphorylation of nuclear proteins bound to Vg1HRE. Involvement of JH III in both induction of Vg1 and control of nuclear proteins binding to Vg1HRE suggest the latter to play an important role in JH signaling.