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The vitellogenin present in the bloodstream undergoes internalization into developing oocytes through the vitellogenin receptor (VgR), a process mediated by receptor-mediated endocytosis. VgR plays a crucial role in facilitating the accumulation of vitellogenin and the maturation of oocytes. In this study, we characterized a Tachypleus tridentatus vitellogenin receptor (TtVgR) gene from the tri-spine horseshoe crab, revealing a length of 1956 bp and encoding 652 amino acid residues with 12 exons. TtVgR has a molecular weight of 64.26 kDa and an isoelectric point of 5.95. Predictions indicate 85 phosphorylation sites and 7 glycosylation sites within TtVgR. Transcriptional analysis demonstrated specific expression of TtVgR in the ovary and yellow connective tissue. TtVgR was identified and distributed in the plasma membrane of oocytes. The siRNA-mediated TtVgR knockdown significantly reduced the transcriptional activity of TtVgR. This depletion induced excessive ROS production, resulting in DNA damage in ovarian primary cells. TUNEL and flow cytometry analyses confirmed ovarian cell apoptosis following TtVgR knockdown, indicating DNA damage in ovarian primary cells. These findings underscore the importance of TtVgR in ovarian cell development, suggesting its potential involvement in vitellogenesis and oocyte maturation. This knowledge may inform innovative breeding strategies and contribute to the sustainable management and conservation of the tri-spine horseshoe crab. [ABSTRACT FROM AUTHOR]

Vitellogenin receptor (VgR) plays a crucial role in oogenesis by mediating endocytosis of vitellogenin and a portion of the yolk proteins in many insect species. However, the function of VgR in minute parasitoid wasps is largely unknown. Here, we applied Trichogramma dendrolimi, a minute egg parasitoid, as a study model to investigate the function of VgR in parasitoids. We developed RNA interference (RNAi) methods based on microinjection of prepupae in T. dendrolimi. RNAi employs nanomaterial branched amphipathic peptide capsules (BAPC) as a carrier for double‐stranded RNA (dsRNA), significantly enhancing delivery efficiency. Also, artificial hosts without medium were used to culture the injected prepupae in vitro. Utilizing these methods, we found that ovarian growth was disrupted after knockdown of TdVgR, as manifested by the suppressed development of the ovariole and the inhibition of nurse cell internalization by oocytes. Also, the initial mature egg load in the ovary was significantly reduced. Notably, the parasitic capacity of the female adult with ovarian dysplasia was significantly decreased, possibly resulting from the low availability of mature eggs. Moreover, ovarian dysplasia in T. dendrolimi caused by VgR deficiency are conserved despite feeding on different hosts. The results confirmed a critical role of TdVgR in the reproductive ability of T. dendrolimi and provided a reference for gene functional studies in minute insects. [ABSTRACT FROM AUTHOR]

Thrips hawaiiensis (Morgan) (Thysanoptera: Thripidae) is a sap-sucking pest that seriously damages several crops and reduces their economic value. Exposure to low concentrations of insecticides may have a sublethal effect on surviving insects. In order to provide a reference for the rational application of emamectin benzoate, its sublethal effects on the development and reproduction of T. hawaiiensis were evaluated. Pupal development time was significantly shorter in T. hawaiiensis treated with sublethal concentrations of emamectin benzoate (LC10 and LC20) than in control. Female adult longevity and female total longevity were significantly longer following LC20 treatment than in the control and LC10 treatment groups. Nevertheless, male adult longevity and male total longevity were significantly shorter in the LC10 treatment group than in the control and LC20 treatment groups. The sublethal concentration of emamectin benzoate (LC20) significantly shortened the preadult stages and the mean generation. Meanwhile, it significantly increased the finite rate of increase, intrinsic rate of increase, and net reproductive rate. The fecundity was significantly higher after LC20 treatment than after LC10 and control treatments. Compared with the control group, the LC10 and LC20 groups of T. hawaiiensis adults showed a significantly higher expression of the vitellogenin (Vg) and vitellogenin receptor (VgR) genes, which played a key role in increasing their fecundity. These findings suggest that short-term exposure to sublethal concentrations of emamectin benzoate may lead to a resurgence and secondary outbreak of T. hawaiiensis infestation. The results have practical applications for the management of this important and noxious pest. [ABSTRACT FROM AUTHOR]

Lipoprotein receptors, including low-density lipoprotein receptor (LDLr) relatives (Lrs) and LDLr-related proteins (Lrps), belong to the LDLr supergene family and participate in diverse physiological functions. In this study, novel sequences of lr and lrp genes expressed in the ovary of the short-finned eel, Anguilla australis, during early gonadal development are presented. The genes encoding the LDLr-like, Lrp1-like, Lrp1b-like, Lrp3, Lrp4-like, Lrp5-like, Lrp6, Lrp10, Lrp11, Lrp12-like, and Lr11-like proteins were found and identified by sequence and structure analysis, in addition to phylogenetic analysis. Genes encoding proteins previously implicated in follicle development and vitellogenin (Vtg) uptake in oviparous vertebrates were also identified, i.e. lr8 (including lr8 + and lr8- variants) and lrp13; their identification was reinforced by conserved synteny with orthologues in other teleost fish. Compared to other lr/lrp genes, the genes encoding Lr8 + , Lr8-, and Lrp13 were highly expressed in ovary during early development, decreasing as oocyte development advanced when induced by hypophysation. Furthermore, lr8 + , lr8-, and lrp13 were dominantly expressed in the ovary when compared with 17 other tissues. Finally, this study successfully detected the expression of both lr8 variants, which showed different expression patterns to those reported in other oviparous vertebrates and provided the first characterisation of Lrp13 in Anguilla sp. We propose that lr8 + , lr8-, and lrp13 encode putative Vtg receptors in anguillid eels. [ABSTRACT FROM AUTHOR]

Vitellogenin receptors (VgRs) transport vitellogenin (Vg) into oocytes, thereby promoting egg growth and embryonic development. VgRs recognize and transport multiple ligands in oviparous animals, but their role in insects is rarely reported. In this study, we investigated whether Bombyx mori VgR (BmVgR) binds and transports lipoprotein-1 (BmLP1) and lipoprotein-7 (BmLP7) of the 30 kDa lipoproteins (30 K proteins), which are essential for egg formation and embryonic development in B. mori. Protein sequence analysis showed BmLP7, similar to reported lipoprotein-3 (BmLP3), contains the cell-penetrating peptides and Cysteine position, while BmLP1 has not. Assays using Spodoptera frugiperda ovary cells (sf9) indicated the direct entry of BmLP7 into the cells, whereas BmLP1 failed to enter. However, co-immunoprecipitation (Co-IP) assays indicated that BmVgR could bind BmLP1. Western blotting and immunofluorescence assays further revealed that over-expressed BmVgR could transport BmLP1 into sf9 cells. Co-IP assays showed that SE11C (comprising LBD1+EGF1+OTC domains of BmVgR) or SE22C (comprising LBD2+EGF2+OTC domains of BmVgR) could bind BmLP1. Over-expressed SE11C or SE22C could also transport BmLP1 into sf9 cells. Western blotting revealed that the ability of SE11C to transport BmLP1 might be stronger than that of SE22C. In the vit mutant with BmVgR gene mutation (vit/vit), SDS-PAGE and western blotting showed the content of BmLP1 in the ovary, like BmVg, was lower than that in the normal silkworm. When transgenic with hsp70 promoter overexpressed BmVgR in the vit mutant, we found that the phenotype of the vit mutant was partly rescued after heat treatment. And contents of BmLP1 and BmVg in vit mutant over-expressed BmVgR were higher than in the vit mutant. We conclude that BmVgR and its two repeat domains could bind and transport BmLP1 into the oocytes of the silkworm, besides BmVg. These results will provide a reference for studying the molecular mechanism of VgR transporting ligands in insects. [ABSTRACT FROM AUTHOR]

Vitellogenin receptor (VgR) is crucial for vitellogenin (Vg) uptake by oocytes. VgR is less known in Arachnida, especially in spiders. Different from only one VgR in an arthropod species, two VgRs, VgR‐1 and VgR‐2, were found in the pond wolf spider, Pardosa pseudoannulata. Both VgRs had the typical domains of the low‐density lipoprotein receptor family except for the absence of the ligand‐binding domain 1 in VgR‐2. Spatiotemporal expression profiles showed that two VgR genes were consistently highly expressed in females and their ovaries, but VgR‐1 was 48‐fold that of VgR‐2 in ovaries. The transcriptional level of VgR‐1 was significantly downregulated by RNAi, but it did not work for VgR‐2 although several trials were performed. Vg‐1 and Vg‐2 might be the ligands of VgR‐1 because their expressions were also decreased in the dsVgR‐1‐treated females. Silencing VgR‐1 prolonged the pre‐oviposition period by 56 h. The expression of VgRs and Vgs were upregulated by juvenile hormones (JHs), which suggested that JHs were the essential factors to vitellogenesis in the spider. The present study revealed the importance of VgR‐1 in the spider oviposition, which will improve the understanding on VgR physiological functions in spiders. [ABSTRACT FROM AUTHOR]

Abstract Background Krüppel homolog 1 (Kr-h1) is a critical transcription factor for juvenile hormone (JH) signaling, known to play a key role in regulating metamorphosis and adult reproduction in insects. Kr-h1 can also be induced by molting hormone 20-hydroxyecdysone (20E), however, the underlying mechanism of 20E-induced Kr-h1 expression remains unclear. In the present study, we investigated the molecular mechanism of Kr-h1 induction by 20E in the reproductive system of a model lepidopteran insect, Bombyx mori. Results Developmental and tissue-specific expression analysis revealed that BmKr-h1 was highly expressed in ovaries during the late pupal and adult stages and the expression was induced by 20E. RNA interference (RNAi)-mediated depletion of BmKr-h1 in female pupae severely repressed the transcription of vitellogenin receptor (VgR), resulting in the reduction in vitellogenin (Vg) deposition in oocytes. BmKr-h1 specifically bound the Kr-h1 binding site (KBS) between − 2818 and − 2805 nt upstream of BmVgR and enhanced the transcription of BmVgR. A 20E cis-regulatory element (CRE) was identified in the promoter of BmKr-h1 and functionally verified using luciferase reporter assay, EMSA and DNA-ChIP. Using pull-down assays, we identified a novel transcription factor B. mori Kr-h1 regulatory protein (BmKRP) that specifically bound the BmKr-h1 CRE and activated its transcription. CRISPR/Cas9-mediated knockout of BmKRP in female pupae suppressed the transcription of BmKr-h1 and BmVgR, resulting in arrested oogenesis. Conclusion We identified BmKRP as a new transcription factor mediating 20E regulation of B. mori oogenesis. Our data suggests that induction of BmKRP by 20E regulates BmKr-h1 expression, which in turn induces BmVgR expression to facilitate Vg uptake and oogenesis.

Vitellogenin receptors (VgRs) have vital roles in reproduction by mediating endocytosis of the vitellogenin (Vg) in oviparous insects. Same as most insect species that possess only one VgR, in this study, a single VgR mRNA (HmVgR) was identified in an important natural enemy ladybeetle, Harmonia axyridis (Pallas). The open reading frame of HmVgR was 5,340 bases encoding a protein of 1,779 amino acids. Bioinformatic analyses showed that HmVgR had conserved domain motifs of low-density lipoprotein receptor family. Based on phylogenetic analysis, HmVgR had highly homologous within the Coleoptera. The transcriptional level of HmVgR was initially detected in the newly emerged female adults, gradually increased from day 3 to day 9, peaked on day 13, and then sharply decreased on day 15. Similar to most insects, HmVgR was expressed specifically in ovarian tissue. Moreover, RNA interference (RNAi) clearly decreased the transcription levels of HmVgR, which blocked the deposition of yolk protein in the ovaries, shortened the ovarian length, and led to negative impacts on reproductive-related parameters (i.e., prolonged preoviposition periods, reduced spawning and depressed hatchability). In sum, these results indicated that HmVgR may be critical for yolk protein deposition of oocytes and can play a key role in reproduction of female adults of H. axyridis. Our results provide conclusive proof for the important roles of HmVgR in fecundity, and establish a basis for further research on its interaction with vitellogenin. Graphical Abstract [ABSTRACT FROM AUTHOR]

Insect pest control can be achieved by the application of RNA interference (RNAi), a key molecular tool in functional genomics. Whereas most RNAi research has focused on insect pests, few studies have been performed on natural enemies. Validating the efficacy of RNAi in natural enemies is crucial for assessing its safety and enabling molecular research on these organisms. Here, we assessed the efficacy of RNAi in the ladybird beetle Eriopis connexa Germar (Coleoptera: Coccinellidae), focusing on genes related to reproduction, such as vitellogenin (Vg) and its receptor (VgR). In the transcriptome of E. connexa, we found one VgR (EcVgR) and two Vg genes (EcVg1 and EcVg2). These genes have been validated by in silico analyses of functional domains and evolutionary relationships. Five-day-old females were injected with 500 ng/µL of a specific double-stranded RNA (dsRNA) (dsEcVg1, dsEcVg2, or dsEcVgR) for RNAi tests, while nonspecific dsRNA (dsGFP or dsAgCE8.1) was used as a control. Interestingly, dsEcVg2 was able to knockdown both Vg genes, while dsEcVg1 could silence only EcVg1. Additionally, the viability of the eggs was significantly reduced when both Vg genes were knocked down at the same time (after treatment with dsEcVg2 or "dsEcVg1+dsEcVg2"). Ultimately, malformed, nonviable eggs were produced when EcVgR was silenced. Interestingly, no dsRNA treatment had an impact on the quantity of eggs laid. Therefore, the feasibility of RNAi in E. connexa has been confirmed, suggesting that this coccinellid is an excellent Neotropical model for molecular research on natural enemies and for studying RNAi nontarget effects., (© 2024 Wiley Periodicals LLC.)

Vitellogenin receptor (VgR) plays a pivotal role in ovarian vitellogenin (Vg) uptake and vertical transmission of pathogenic microbes and Wolbachia symbionts. However, the regulatory mechanisms of VgR action as an endocytic receptor and translocation from oocyte cytoplasm to the membrane remain poorly understood. Here, by using the migratory locust Locusta migratoria as a model system, we report that juvenile hormone (JH) promotes VgR phosphorylation at Ser1361 in the second EGF-precursor homology domain. A signaling cascade including GPCR, PLC, extracellular calcium, and PKC-ι is involved in JH-stimulated VgR phosphorylation. This posttranslational regulation is a prerequisite for VgR binding to Vg on the external surface of the oocyte membrane and subsequent VgR/Vg endocytosis. Acidification, a condition in endosomes, induces VgR dephosphorylation along with the dissociation of Vg from VgR. Phosphorylation modification is also required for VgR recycling from oocyte cytoplasm to the membrane. Additionally, VgR phosphorylation and its requirement for Vg uptake and VgR recycling are evolutionarily conserved in other representative insects including the cockroach Periplaneta americana and the cotton bollworm Helicoverpa armigera. This study fills an important knowledge gap of low-density lipoprotein receptors in posttranslational regulation, endocytosis, and intracellular recycling. [ABSTRACT FROM AUTHOR]

Abstract Background Vitellogenin (Vg), a key molecule for oocyte development synthesized in the fat body during blood-feeding, is released into the hemolymph and then taken into the oocytes via Vg receptor (VgR) in ticks. Previously, we showed that VgR mRNA is expressed in the ovary at the adult stage of parthenogenetic Haemaphysalis longicornis ticks and its expression increases after blood-feeding. However, intracellular localization of VgR mRNA and protein at each developmental stage of oocytes during oogenesis remains largely unclear. Methods mRNA and protein expression profiles of H. longicornis VgR (HlVgR) in the oocytes from the unfed to oviposition periods were analyzed by real-time PCR, in situ hybridization, and immunostaining. To elucidate the timing of the onset of Vg uptake, RNA interference (RNAi)-mediated gene silencing of HlVgR was performed. Results In situ hybridization revealed that HlVgR mRNA was detected in the cytoplasm of stage I-III oocytes, and weaker positive signals for HlVgR mRNA were found in the cell periphery of stage IV and V oocytes. Likewise, HlVgR protein was detected by immunostaining in the cytoplasm of stage I-III oocytes and in the cell periphery of stage IV and V oocytes. Each developmental stage of the oocytes showed distinct patterns of mRNA and protein expression of HlVgR. Moreover, RNAi of HlVgR caused delayed or arrested development in the oocytes. The ovaries of control ticks showed all developmental stages of oocytes, whereas stage I-III oocytes were found in the ovaries of HlVgR-RNAi ticks at 5 days after engorgement. Conclusions These results suggest that active uptake of Vg is required for development from stage III to stage IV during oogenesis. Our data clearly revealed an apparent shift in the intracellular localization of VgR for both mRNA and protein level in oocytes during oogenesis.

Abstract Background Rhipicephalus microplus is an efficient biological vector of Babesia bovis, a causative agent of bovine babesiosis. Babesia bovis is passed transovarially to the next generation of ticks, which then transmit the parasite to naïve animals. Due to the importance of the R. microplus ovary for tick reproduction and transmission of B. bovis, we investigated the hypothesis that silencing vitellogenin receptor gene expression in the ovary during tick feeding on B. bovis-infected cattle would affect parasite transmission to the next generation of ticks. Results Silencing expression of the vitellogenin receptor in the ovary by RNA interference, resulted in reduced tick fertility. We observed reduced egg production (i.e. reduced weight of eggs), a lower rate of embryonic development, and a reduction in hatching. Analysis of individual larvae by PCR confirmed that RNAi mediated downregulation of the R. microplus vitellogenin receptor and also interfered with transovarial transmission of B. bovis. None of the larvae (0/58) from the RmVgR dsRNA-injected group were PCR-positive, whereas 12% (7/58) and 17% (10/58) of larvae from the non-injected and buffer-injected control groups, respectively, were infected with B. bovis. Conclusions The combined effects of reduced fecundity and reduced infection in surviving larvae resulting from silencing indicate that vitellogenin receptor is essential for tick reproduction and may play a vital role in B. bovis transmission.

BACKGROUND: Aedes albopictus is an important vector with an extensive worldwide distribution. Only female mosquitoes play a significant role in the transmission of pathogens. Doublesex (dsx) is a central nexus gene in the insect somatic sex determination hierarchy. RESULTS: In this study, we characterized the full-length sex-specific splicing forms of the Ae. albopictus dsx (Aalbdsx) gene. Then, we identified 15 direct target genes of DSX in adult females using digital gene expression combined with quantitative real-time polymerase chain reaction (qPCR) by performing a chromatin immunoprecipitation (ChIP) assay with specific DSX antibodies. Knockdown of Aalbdsx suppressed ovarian development and decreased the transcript levels of the Aalbdsx target vitellogenin receptor (VgR) gene, whereas vitellogenin (Vg) expression showed an increase in the fat body. Genes in the major Vg regulatory pathway were also up-regulated. CONCLUSION: Our results suggest that both Vg and VgR are direct target genes of Aalbdsx and that direct regulation of Aalbdsx on VgR is indispensable for ovarian development in Ae. albopictus, which not only provides a reference for the further elucidation of the evolutionarily conserved role of dsx in Ae. albopictus sexual differentiation but also reveals potential molecular targets for application to the development of sterile male mosquitoes to be released for vector control. [ABSTRACT FROM AUTHOR]

Background: Krüppel homolog 1 (Kr-h1) is a critical transcription factor for juvenile hormone (JH) signaling, known to play a key role in regulating metamorphosis and adult reproduction in insects. Kr-h1 can also be induced by molting hormone 20-hydroxyecdysone (20E), however, the underlying mechanism of 20E-induced Kr-h1 expression remains unclear. In the present study, we investigated the molecular mechanism of Kr-h1 induction by 20E in the reproductive system of a model lepidopteran insect, Bombyx mori. Results: Developmental and tissue-specific expression analysis revealed that BmKr-h1 was highly expressed in ovaries during the late pupal and adult stages and the expression was induced by 20E. RNA interference (RNAi)-mediated depletion of BmKr-h1 in female pupae severely repressed the transcription of vitellogenin receptor (VgR), resulting in the reduction in vitellogenin (Vg) deposition in oocytes. BmKr-h1 specifically bound the Kr-h1 binding site (KBS) between − 2818 and − 2805 nt upstream of BmVgR and enhanced the transcription of BmVgR. A 20E cis-regulatory element (CRE) was identified in the promoter of BmKr-h1 and functionally verified using luciferase reporter assay, EMSA and DNA-ChIP. Using pull-down assays, we identified a novel transcription factor B. mori Kr-h1 regulatory protein (BmKRP) that specifically bound the BmKr-h1 CRE and activated its transcription. CRISPR/Cas9-mediated knockout of BmKRP in female pupae suppressed the transcription of BmKr-h1 and BmVgR, resulting in arrested oogenesis. Conclusion: We identified BmKRP as a new transcription factor mediating 20E regulation of B. mori oogenesis. Our data suggests that induction of BmKRP by 20E regulates BmKr-h1 expression, which in turn induces BmVgR expression to facilitate Vg uptake and oogenesis. [ABSTRACT FROM AUTHOR]

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Besides the function of preventing metamorphosis in insects, the juvenile hormone (JH) plays a role in female reproduction; however, the underlying mechanism is largely unknown. The methoprene-tolerant (Met) protein belongs to a family of basic helix-loop-helix–Per-Arnt-Sim (bHLH-PAS) transcription factors and functions as the JH intracellular receptor. In this study, two full length cDNAs encoding Met (CsMet1 and CsMet2) were isolated from the rice stem borer, Chilo suppressalis. Structural analysis revealed that both CsMet1 and CsMet2 exhibited typical bHLH, PAS-A, PAS-B, and PAC (PAS C terminal motif) domains. Comparative analysis of transcript level using reverse transcription-quantitative PCR (RT-qPCR) revealed that CsMet1 was predominant in almost all examined developmental stages and tissues. Treatment with methoprene in vivo induces the transcription of both CsMet1 and CsMet2. Notably, injection of dsCsMet1 and dsCsMet2 suppressed the expression levels of vitellogenin (CsVg) and Vg receptor (CsVgR). These findings revealed the potential JH signaling mechanism regulating C. suppressalis reproduction, and provided evidence that RNAi-mediated knockdown of Met holds great potential as a control strategy of C. suppressalis.

Oocyte maturation and ovarian development are sequentially coordinated events critical to reproduction. In the ovaries of adult oviparous animals such as birds, bony fish, insects, and crustaceans, vitellogenin receptor (VgR) is a plasma membrane receptor that specifically mediates vitellogenin (Vg) transport into oocytes. Accumulation of Vg drives sexual maturation of the female crustaceans by acting as a pivotal regulator of nutritional accumulation within oocytes, a process known as vitellogenesis. However, the mechanisms by which VgR mediates vitellogenesis are still not fully understood. In this study, we first identified a unique VgR (Lv-VgR) and characterized its genomic organization and protein structural domains in Litopenaeus vannamei, a predominant cultured shrimp species worldwide. This newly identified Lv-VgR phylogenetically forms a group with VgRs from other crustacean species within the arthropod cluster. Duplicated LBD/EGFD regions are found exclusively among arthropod VgRs but not in paralogs from vertebrates and nematodes. In terms of expression patterns, Lv-VgR transcripts are specifically expressed in ovaries of female shrimps, which increases progressively during ovarian development, and rapidly declines toward embryonic development. The cellular and subcellular locations were For analyzed by in situ hybridization and immunofluorescence, respectively. The Lv-VgR mRNA was found to be expressed in the oocytes of ovaries, and Lv-VgR protein was found to localize in the cell membrane of maturing oocytes while accumulation of the ligand Vg protein assumed an even cytoplasmic distribution. Silencing of VgR transcript expression by RNAi was effective for stunting ovarian development. This present study has thus provided new insights into the regulatory roles of VgR in crustacean ovarian development.

At the onset of puberty, ovarian follicles become competent to incorporate large amounts of vitellogenin (Vtg). Using an RNAseq-based approach, transcriptomes from pre-vitellogenic (PV) and early vitellogenic (EV) ovaries from wild-caught eel, Anguilla australis, were compared to investigate the expression of specific genes encoding cell junction proteins that could be involved in regulating Vtg uptake. Partial support was found for the mechanical barrier hypothesis proposing that the access of Vtg to the oolemma is restricted by a tight junction (TJ) network within the granulosa cell layer, which changes between the PV and EV stage. Among 25 genes encoding TJ-constituting proteins, five were down-regulated and two were up-regulated. A chemical barrier hypothesis stating that gap junctions (GJs) are involved in modulating Vtg uptake was not supported, as only five GJs were found to be expressed in the ovary with no significant changes in expression between stages. Furthermore, the endocytic pathway was found to be up-regulated during the PV-EV transition. Finally, the study showed that gene expression patterns may help identify suitable candidates involved in the regulation of Vtg uptake, and provided novel sequence data for A. australis, including putative Vtg receptors corresponding to Lr8 and Lrp13 members of the low-density lipoprotein receptor family.

The vitellogenin receptor (Vgr), which is specific for vitellogenin (Vtg), recognises and transports Vtg into the ovaries. Accumulating evidence suggests that Vtg also performs an immune defence function and plays critical roles in innate immunity in oviparous animals. However, whether Vgr is involved in innate immunity in the Chinese mitten crab (Eriocheir sinensis) is unknown. In this study, we obtained a 3009 nucleotide partial cDNA of the E. sinensis vitellogenin receptor gene (Es-vgr) encoding an open reading frame of 1003 amino acid residues. Bioinformatics analysis showed that the domains of Es-vgr were conserved during evolution. Quantitative real-time PCR and western blotting revealed that the highest Es-vgr expression levels occurred in the ovary, and expression was specific. Comparison of the expression levels of Es-vgr and the Vtg gene (Es-vtg1) at different ovary developmental stages suggested that there may be some regulatory relationship between them. Bacterial challenge induced high-level expression of antimicrobial peptide genes and reduced Es-vgr expression in ovaries, resulting in massive accumulation of Vtg in the hemolymph. The survival rate of crabs increased significantly after injection with recombinant Es- vtg1 protein following bacterial infection. Collectively, these results demonstrate that Es-vgr plays critical roles in antimicrobial function by regulating the accumulation of Vtg in the hemolymph. • The expression of Vtg and Vgr were both changed dynamically in different developmental stages. • Down-regulated Vgr transcription may cause Vtg accumulation in hemolymph post bacterial challenge. • Vtg has critical role in protect crab from bacterial infection. [ABSTRACT FROM AUTHOR]

The almond moth is an important pest of date fruits worldwide. The females produce several eggs; however, role of vitellogenin receptor (VgR) in oocyte development by mediating endocytosis of major yolk protein precursor Vg remains yet unexplored. To investigate the role of vitellogenin receptor (VgR) in reproduction, Cadra cautella vitellogenin receptor (CcVgR) transcript was obtained using rapid amplification of cDNA ends‐polymerase chain reaction. Expression analysis of CcVgR was performed using reverse transcriptase and quantitative polymerase chain reaction (qPCR) in different developmental stages. RNA interference was performed by injecting CcVgR‐based double‐stranded (ds)RNA at different exposure times. The results revealed that CcVgR is 5421 bp long, encoded 1807 amino acid, belongs to low‐density lipoprotein receptor superfamily and contains all conserved domains. Expression analysis confirmed that CcVgR is sex‐specific and starts to express in female larvae on day 19. Additionally, RNA interference (RNAi) of CcVgR‐based dsRNA inhibited CcVgR expression up to 83% after 72 h, reduced fecundity and hatchability, and confirmed involvement of CcVgR in C. cautella reproduction. This report provides a basis for gene silencing in this species, and proposes RNAi technology potential for pest management. [ABSTRACT FROM AUTHOR]

Besides the function of preventing metamorphosis in insects, the juvenile hormone (JH) plays a role in female reproduction; however, the underlying mechanism is largely unknown. The methoprene-tolerant (Met) protein belongs to a family of basic helix-loop-helix–Per-Arnt-Sim (bHLH-PAS) transcription factors and functions as the JH intracellular receptor. In this study, two full length cDNAs encoding Met (CsMet1 and CsMet2) were isolated from the rice stem borer, Chilo suppressalis. Structural analysis revealed that both CsMet1 and CsMet2 exhibited typical bHLH, PAS-A, PAS-B, and PAC (PAS C terminal motif) domains. Comparative analysis of transcript level using reverse transcription-quantitative PCR (RT-qPCR) revealed that CsMet1 was predominant in almost all examined developmental stages and tissues. Treatment with methoprene in vivo induces the transcription of both CsMet1 and CsMet2. Notably, injection of dsCsMet1 and dsCsMet2 suppressed the expression levels of vitellogenin (CsVg) and Vg receptor (CsVgR). These findings revealed the potential JH signaling mechanism regulating C. suppressalis reproduction, and provided evidence that RNAi-mediated knockdown of Met holds great potential as a control strategy of C. suppressalis. [ABSTRACT FROM AUTHOR]

Oocyte maturation and ovarian development are sequentially coordinated events critical to reproduction. In the ovaries of adult oviparous animals such as birds, bony fish, insects, and crustaceans, vitellogenin receptor (VgR) is a plasma membrane receptor that specifically mediates vitellogenin (Vg) transport into oocytes. Accumulation of Vg drives sexual maturation of the female crustaceans by acting as a pivotal regulator of nutritional accumulation within oocytes, a process known as vitellogenesis. However, the mechanisms by which VgR mediates vitellogenesis are still not fully understood. In this study, we first identified a unique VgR (Lv-VgR) and characterized its genomic organization and protein structural domains in Litopenaeus vannamei , a predominant cultured shrimp species worldwide. This newly identified Lv-VgR phylogenetically forms a group with VgRs from other crustacean species within the arthropod cluster. Duplicated LBD/EGFD regions are found exclusively among arthropod VgRs but not in paralogs from vertebrates and nematodes. In terms of expression patterns, Lv-VgR transcripts are specifically expressed in ovaries of female shrimps, which increases progressively during ovarian development, and rapidly declines toward embryonic development. The cellular and subcellular locations were For analyzed by in situ hybridization and immunofluorescence, respectively. The Lv-VgR mRNA was found to be expressed in the oocytes of ovaries, and Lv-VgR protein was found to localize in the cell membrane of maturing oocytes while accumulation of the ligand Vg protein assumed an even cytoplasmic distribution. Silencing of VgR transcript expression by RNAi was effective for stunting ovarian development. This present study has thus provided new insights into the regulatory roles of VgR in crustacean ovarian development. [ABSTRACT FROM AUTHOR]

Vitellogenin receptors (VgRs) play crucial roles in oogenesis by mediating endocytosis of vitellogenin and other nutrients in ovipara. We conducted small RNA sequencing and screening with a luciferase reporter system, and found that bmo-miR-2739 and a novel miRNA (novel-miR-167) coordinately regulate the expression of VgR in Bombyx mori (BmVgR). Further analyses suggested that these two miRNAs direct target repression by binding directly to the BmVgR 3' untranslated region. Forced expression of either miRNA using the piggyBac system blocked vitellogenin (Vg) transport and retarded ovariole development. Antagomir silencing of bmo-miR-2739 or novelmiR- 167 resulted in increased amounts of BmVgR protein in the ovaries and BmVgR mRNA in the fat body. This evidence, combined with spatiotemporal expression profiles, revealed that these two miRNAs function together to fine-tune the amount of BmVgR protein for ovarian development. Additionally, novel-miR-167 was mainly responsible for the post-transcriptional repression of BmVgR in nonovarian tissues. The results of this study contribute to our understanding of the function of miRNAs during ovarian development of a lepidopteran and suggest a new strategy for controlling insect reproduction. [ABSTRACT FROM AUTHOR]

Vitellogenin receptors (VgRs) play critical roles in egg formation by transporting vitellogenin (Vg) into oocytes in insects. Although the function of VgR in insects is well studied, the transcriptional regulation of this gene is still unclear. Here, we cloned the promoter of the VgR gene from Bombyx mori (BmVgR), and predicted many POU cis-response elements (CREs) in its promoter. Electrophoretic mobility shift and chromatin immunoprecipitation assays showed that the POU transcription factor POU-M2 bound directly to the CREs of the promoter. Overexpression of POU-M2 in an ovarian cell line (BmNs) enhanced BmVgR transcription and promoter activity detected by quantitative reverse transcription PCR and luciferase reporter assays. Analyses of expression patterns indicated that POU-M2 was expressed in ovary at day two of wandering stage initially, followed by BmVgR. RNA interference of POU-M2 significantly reduced the transcription of BmVgR in ovary and egg-laying rate. Our results suggest a novel function for the POU factor in silkworm oogenesis by its involvement in BmVgR regulation and expands the understanding of POU factors in insect VgR expression. [ABSTRACT FROM AUTHOR]

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<0.05), while the hatching rate of eggs not significantly (P>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. [ABSTRACT FROM AUTHOR]

Clathrin plays an important role in arthropods, but its function in ticks has not been explored. Here, we describe the molecular characteristics of the clathrin heavy chain of the tick Rhipicephalus haemaphysaloides and its effects on yolk development. The open reading frame of the clathrin heavy chain (Chc) (Rh-Chc) gene consists of 5103 nucleotides encoding 670 amino acids, which is most closely related to that of Ixodes scapularis and relatively close to Homo sapiens and Drosophila melanogaster. Real-time qPCR revealed that Rh-Chc was expressed at all developmental stages and organs. After Rh-Chc is silenced, ticks did not feed and mortality rate was 100%. Moreover, Rh-Chc co-localized with Vitellogenin receptor (VgR) on oocyte membrane. Immunofluorescence showed that the expression of Vitellogenin (Vg) (Rh-Vg) was also closely related to Rh-Chc. Immunofluorescence showed that the expression of Vg was also closely related to Rh-Chc, Rh-Chc silencing slowed the development of oocytes in tick, and culture of ovary in vitro silenced Rh-Chc, the development of oocytes in ticks also slowed down. Overall, the results of this study indicated that Rh-Chc is a vital gene in the tick R. haemaphysaloides that plays an important role in its growth, development, and reproduction. [ABSTRACT FROM AUTHOR]

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.

Agasicles hygrophila is an effective biological control agent for the invasive weed Alternanthera philoxeroides, and because of this it has been introduced to many parts of the world where A. philoxeroides is a problem. Despite this, there are no reports at present about the reproduction of this important insect. Vitellogenin receptors (VgRs) belong to the superfamily of low-density lipoprotein receptors (LDLRs). One of the roles of VgRs is to regulate the absorption of yolk protein in insects. In this study, the full length vitellogenin receptor gene (AhVgR) from A. hygrophila was sequenced and found to encode a predicted protein of 1,642 amino acids. Sequence analysis of AhVgR revealed that it contains conserved structural motifs common to LDLR family members, and a phylogenetic analysis placed AhVgR as a separate group among the order Coleoptera. AhVgR was found to be specifically expressed in ovarian tissues, and it is first transcribed in the newly-emerged females. The expression patterns are consistent with VgR genes in other insects. RNA interference (RNAi)-mediated suppression of AhVgR gene expression in adult A. hygrophila females inhibited yolk protein deposition in the ovaries, shortened the ovariole, drastically reduced egg production, and ultimately led to a decrease in fecundity. In summary, our work shows that AhVgR is critical for transporting Vg into the oocytes and plays an important role in A. hygrophila reproduction.

While much effort has been put into understanding vitellogenesis in insects and other organisms, much less is known of this process in ticks. There are several steps that facilitate yolk formation in developing oocytes of which the vitellogenin receptor (VgR) is a key component. The tick VgR binds vitellogenin (Vg) circulating in the hemolymph to initiate receptor-mediated endocytosis and its transformation into vitellin (Vn). The conversion of Vg into Vn, the final form of the yolk protein, occurs inside oocytes of the female tick ovary. Vn is critical to tick embryos since it serves as the nutritional source for their development, survival, and reproduction. Recent studies also suggest that pathogenic microbes, i.e., Babesia spp., that rely on ticks for propagation and dissemination likely “hitchhike” onto Vg molecules as they enter developing oocytes through the VgR. Suppressing VgR messenger RNA synthesis via RNA interference (RNAi) completely blocked Babesia spp. transmission into developing tick oocytes, thereby inhibiting vertical transmission of these pathogenic microbes from female to eggs. To date, VgRs from only four tick species, Dermacentor variabilis, Rhipicephalus microplus, Amblyomma hebraeum, and Haemaphysalis longicornis, have been fully sequenced and characterized. In contrast, many more VgRs have been described in various insect species. VgR is a critical component in egg formation and maturation that can serve as a precise target for tick control. However, additional research will help identify unique residues within the receptor that are specific to ticks or other arthropod disease vectors while avoiding cross-reactivity with non-target species. Detailed knowledge of the molecular structure and functional role of tick VgRs will enable development of novel vaccines to control ticks and tick-borne diseases.

Adipokinetic hormones (AKHs) are well known to mobilize lipids and carbohydrates for energy-consuming activities in insects. These neuropeptides exert their functions by interacting with AKH receptors (AKHRs) located on the plasma membrane of fat body cells, which regulates energy mobilization by stimulating lipolysis of triacylglycerols (TAG) to diacylglycerols (DAG) and conversion of glycogen into trehalose. Here, we investigated the roles of AKH/AKHR signaling system in trehalose metabolism and vitellogenesis during female reproduction in the brown planthopper, Nilaparvata lugens. Knockdown of AKHR expression by RNA interference (RNAi) resulted in a decrease of the circulating trehalose in hemolymph and significantly increased levels of two trehalases in fat bodies, indicating that the modulation of hemolymph trehalose levels by AKHR may be mediated by regulating trehalose degradation. In addition, adult females that had been injected with double-stranded RNA (dsRNA) for AKHR exhibited delayed oocyte maturation, prolonged pre-oviposition period, as well as decline in egg number and reduction in fecundity. Considering that these phenotypes resulting from AKHR silencing are similar to those of vitellogenin receptor (VgR) RNAi, we further analyzed a possible connection between AKHR and vitellogenesis. Knockdown of AKHR showed no effects on the Vg synthesis in fat bodies, whereas it significantly reduced the levels of VgR in ovaries. With RNAi-females, we observed an increase of Vg accumulation in hemolymph and a decrease of Vg deposition in ovaries. Moreover, the decrease in VgR expression and Vg incorporation by developing oocytes could be partially rescued by injection of trehalose into AKHR RNAi females. The present study has implicated trehalose in the AKH/AKHR signaling-mediated control of reproduction and provided new insight into mechanisms of AKH/AKHR regulation of trehalose metabolism in insect vitellogenesis, oocyte maturation and fecundity.

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. [ABSTRACT FROM AUTHOR]

Accumulation of nonylphenol (NP) in hepatopancreas, gonad, eyestalk, and muscle of freshwater prawn Macrobrachium rosenbergii following 72 h exposure to 100 µg/L NP, and depuration of NP in these tissues at 0.5–192 h post exposure were examined. We also examined the expressions of vitellogenin (Vg) and vitellogenin receptor (VgR) of prawn following 0–20 days exposure to 0, 1, 10, and 100 µg/L NP. NP accumulation in hepatopancreas and gonad with high concentration, and low concentration in muscle, but depurated faster in eyestalk and muscle. The expressions of vitellogenin (Vg) and vitellogenin receptor (VgR) increased directly with dose and time. In conclusion, NP accumulated significantly in gonad together with high Vg and VgR expressions, and depurated slow in hepatopancreas and gonad when prawns were removed back to control water. The induction of Vg and VgR under NP exposure might be a stress response in M. rosenbergii. [ABSTRACT FROM AUTHOR]

Agasicles hygrophila is an effective biological control agent for the invasive weed Alternanthera philoxeroides , and because of this it has been introduced to many parts of the world where A. philoxeroides is a problem. Despite this, there are no reports at present about the reproduction of this important insect. Vitellogenin receptors (VgRs) belong to the superfamily of low-density lipoprotein receptors (LDLRs). One of the roles of VgRs is to regulate the absorption of yolk protein in insects. In this study, the full length vitellogenin receptor gene (AhVgR) from A. hygrophila was sequenced and found to encode a predicted protein of 1,642 amino acids. Sequence analysis of AhVgR revealed that it contains conserved structural motifs common to LDLR family members, and a phylogenetic analysis placed AhVgR as a separate group among the order Coleoptera. AhVgR was found to be specifically expressed in ovarian tissues, and it is first transcribed in the newly-emerged females. The expression patterns are consistent with VgR genes in other insects. RNA interference (RNAi)-mediated suppression of AhVgR gene expression in adult A. hygrophila females inhibited yolk protein deposition in the ovaries, shortened the ovariole, drastically reduced egg production, and ultimately led to a decrease in fecundity. In summary, our work shows that AhVgR is critical for transporting Vg into the oocytes and plays an important role in A. hygrophila reproduction. [ABSTRACT FROM AUTHOR]

While much effort has been put into understanding vitellogenesis in insects and other organisms, much less is known of this process in ticks. There are several steps that facilitate yolk formation in developing oocytes of which the vitellogenin receptor (VgR) is a key component. The tick VgR binds vitellogenin (Vg) circulating in the hemolymph to initiate receptor-mediated endocytosis and its transformation into vitellin (Vn). The conversion of Vg into Vn, the final form of the yolk protein, occurs inside oocytes of the female tick ovary. Vn is critical to tick embryos since it serves as the nutritional source for their development, survival, and reproduction. Recent studies also suggest that pathogenic microbes, i.e., Babesia spp., that rely on ticks for propagation and dissemination likely "hitchhike" onto Vg molecules as they enter developing oocytes through the VgR. Suppressing VgR messenger RNA synthesis via RNA interference (RNAi) completely blocked Babesia spp. transmission into developing tick oocytes, thereby inhibiting vertical transmission of these pathogenic microbes from female to eggs. To date, VgRs from only four tick species, Dermacentor variabilis , Rhipicephalus microplus , Amblyomma hebraeum , and Haemaphysalis longicornis , have been fully sequenced and characterized. In contrast, many more VgRs have been described in various insect species. VgR is a critical component in egg formation and maturation that can serve as a precise target for tick control. However, additional research will help identify unique residues within the receptor that are specific to ticks or other arthropod disease vectors while avoiding cross-reactivity with non-target species. Detailed knowledge of the molecular structure and functional role of tick VgRs will enable development of novel vaccines to control ticks and tick-borne diseases. [ABSTRACT FROM AUTHOR]

BACKGROUND The small brown plant hopper (SBPH), Laodelphax striatellus Fallén, is one of the most destructive pests on rice. This pest transmits rice stripe virus (RSV) both horizontally and vertically, leading to major yield and economic losses in rice production. However, the way that RSV particles enter oocytes of SBPH remains largely unknown. Thus, identification of key factors involved in the interaction between SBPH and RSV in the ovary is crucial. RESULTS: Transcriptome of non‐viruliferous (NV) or high viruliferous (HV) SBPH ovaries at 24 and 48 h of emergence was sequenced. Differentially expressed genes analysis showed that vitellogenin receptor was significantly highly expressed in the ovary of the HV SBPH strains compared to NV strains. Quantitative real‐time polymer chain reaction showed that the vitellogenin receptor in L. striatellus (LsVgR) was highly expressed in the ovaries of female adults and maintained a high level of expression at the early stage of ovary development. By using RNA interference, the expression of LsVgR in the ovaries of the HV strain was significantly decreased by 98.1%. RSV titer was reduced by 60.9% as quantified by viral RNA3 intergenic region and the transcripts of nucleocapsid protein gene (CP) reduced by 46.3%. The numbers of offspring hatched were significantly reduced in dsRNA‐treated groups. The transcripts of CP were not affected by silencing LsVgR, whereas the abundance of RNA‐dependent RNA polymerase increased by 15‐fold in the member of surviving progenies. CONCLUSION: Our results suggest that vitellogenin receptor participates in regulating RSV replication during oogenesis. © 2018 Society of Chemical Industry This study investigates factors involved in the interaction between rice stripe virus and Laodelphax striatellus in ovaries, showing that vitellogenin receptor is involved in regulating RSV replication during oogenesis. [ABSTRACT FROM AUTHOR]

Abstract The vitellogenin receptor (VgR) is highly expressed in the ovaries where it is responsible for vitellogenin (Vg) deposition during oogenesis in insects. Therefore, identifying the VgR of insect pests, and understanding the mechanism regulating its expression, could lead to the development of pest management strategies based on disrupting reproduction. We cloned and identified VgR in the cabbage beetle, Colaphellus bowringi , which is a serious pest of cruciferous vegetables in Asia. The regulation of VgR transcription by juvenile hormone (JH) was also investigated. The results show that C. bowringi VgR cDNA contains an open reading frame of 5310 bp encoding 1769 amino acid residues. Protein domain prediction indicates that C. bowringi VgR belongs to the LDLR gene superfamily, having the same group of structural domains that has been well characterized in other insects. VgR mRNA was highly expressed in the ovaries of reproductive female cabbage beetles. Knockdown of VgR reduced yolk deposition in the ovaries, increased the accumulation of Vg proteins in the hemolymph and decreased the transcription of Vg1 and Vg2 in the fat body. RNA interference and hormone challenge experiments showed that JH induced VgR transcription via the JH intracellular receptor methoprene-tolerant (Met) and the JH-responsive transcription factor Krüppel homolog 1 (Kr-h1). Our results suggest that there is a feedback loop between VgR transcription in the ovaries and Vg transcription in the fat body. JH acting through Met-Kr-h1 pathway induces the transcription of the VgR that is essential for Vg uptake and reproductive development. These findings not only reveal the potential JH signaling mechanism regulating VgR transcription, but may also contribute to the development of pest control strategies based on disrupting endocrine-regulated reproduction. Graphical abstract Unlabelled Image Highlights • Colaphellus bowringi VgR mRNA level was higher in the ovaries of reproductive females compared to diapause females. • Knockdown of VgR reduced yolk deposition in the ovaries and increased the accumulation of Vg proteins in the hemolymph. • There was a feedback loop between VgR transcription in the ovaries and Vg transcription in the fat body. • JH induced VgR transcription via the Met-Kr-h1 pathway. [ABSTRACT FROM AUTHOR]

Adipokinetic hormones (AKHs) are well known to mobilize lipids and carbohydrates for energy-consuming activities in insects. These neuropeptides exert their functions by interacting with AKH receptors (AKHRs) located on the plasma membrane of fat body cells, which regulates energy mobilization by stimulating lipolysis of triacylglycerols (TAG) to diacylglycerols (DAG) and conversion of glycogen into trehalose. Here, we investigated the roles of AKH/AKHR signaling system in trehalose metabolism and vitellogenesis during female reproduction in the brown planthopper, Nilaparvata lugens. Knockdown of AKHR expression by RNA interference (RNAi) resulted in a decrease of the circulating trehalose in hemolymph and significantly increased levels of two trehalases in fat bodies, indicating that the modulation of hemolymph trehalose levels by AKHR may be mediated by regulating trehalose degradation. In addition, adult females that had been injected with double-stranded RNA (dsRNA) for AKHR exhibited delayed oocyte maturation, prolonged pre-oviposition period, as well as decline in egg number and reduction in fecundity. Considering that these phenotypes resulting from AKHR silencing are similar to those of vitellogenin receptor (VgR) RNAi, we further analyzed a possible connection between AKHR and vitellogenesis. Knockdown of AKHR showed no effects on the Vg synthesis in fat bodies, whereas it significantly reduced the levels of VgR in ovaries. With RNAi-females, we observed an increase of Vg accumulation in hemolymph and a decrease of Vg deposition in ovaries. Moreover, the decrease in VgR expression and Vg incorporation by developing oocytes could be partially rescued by injection of trehalose into AKHR RNAi females. The present study has implicated trehalose in the AKH/AKHR signaling-mediated control of reproduction and provided new insight into mechanisms of AKH/AKHR regulation of trehalose metabolism in insect vitellogenesis, oocyte maturation and fecundity. [ABSTRACT FROM AUTHOR]

Abstract Lambda-cyhalothrin has long been recommended as an effective insecticide to control the oriental armyworm, Mythimna separata (Walker), a notorious migratory pest of agricultural plants. Previous researches have suggested that survival, development, and reproduction of insects are influenced by sublethal concentrations of insecticides. However, the effects of sublethal concentrations of lambda-cyhalothrin on M. separata are less known. In this study, we determined the toxicity and effects of LC 20 and LC 50 concentration of lambda-cyhalothrin on development and reproduction of M. separata. Results indicate that LC 20 of lambda-cyhalothrin tends to decrease the life traits of M. separate , with a shortening larvae period of offspring and oviposition period, whereas LC 50 of lambda-cyhalothrin stimulates daily maximal fecundity and forwards the oviposition peak, suggesting a stimulation of reproduction by LC 50 of lambda-cyhalothrin. The M. separata population was increased by an LC 50 concentration of lambda-cyhalothrin, resulting in a net reproductive rate (R 0) and intrinsic rate of increase (r m) significantly higher than that of the control. Transcripts of vitellogenin (MsVg) and vitellogenin receptor (MsVgR) genes were suppressed at day 1 after emergence of moth which developed from the larvae exposed to LC 20 and LC 50 of lambda-cyhalothrin, but were significant induced when the moth begin to lay eggs (day 4), with a more remarkable induction by LC 50 of lambda-cyhalothrin than those of LC 20 of lambda-cyhalothrin. Our results indicate that the observed stimulation of reproduction is therefore the results of up-regulation of MsVg and MsVgR by LC 50 of lambda-cyhalothrin. Graphical abstract Unlabelled Image Highlights • The toxicity of lambda-cyhalothrin on Mythimna separata was determined. • LC 20 of lambda-cyhalothrin decreases the life traits of M. separate. • LC 50 of lambda-cyhalothrin stimulates reproduction on moth M. separata. • LC 50 of lambda-cyhalothrin up-regulates the expression of vitellogenin and vitellogenin receptor gene in M. separata. [ABSTRACT FROM AUTHOR]

Abstract Pesticides risk assessments have traditionally focused on the effects on standard parameters, such as mortality, reproduction and development. However, one of the first signs of adverse effects that occur in organisms exposed to stress conditions is an alteration in their genomic expression, which is specific to the type of stress, sensitive to very low contaminant concentrations and responsive in a few hours. The aim of the present study was to evaluate the single and binary mixture toxicity of commercial products of abamectin (Kraft® 36 EC) and difenoconazole (Score® 250 EC) to Folsomia candida. Laboratory toxicity tests were conducted to access the effects of these pesticides on springtail survival, reproduction and gene expression. The reproduction assays gave EC 50 and EC 10 values, respectively, of 6.3 and 1.4 mg a.s./kg dry soil for abamectin; 1.0 and 0.12 mg a.s./kg dry soil for Kraft® 36 EC; and 54 and 23 mg a.s./kg dry soil for Score® 250 EC. Technical difenoconazole did not have any effect at the concentrations tested. No significant differences in gene expression were found between the abamectin concentrations tested (EC 10 and EC 50) and the solvent control. Exposure to Kraft® 36 EC, however, significantly induced Cyp6 expression at the EC 50 level, while VgR was significantly downregulated at both the EC 10 and EC 50. Exposure to the simple pesticide mixture of Kraft® 36 EC + Score® 250 EC caused significant up regulation of ABC transporter, and significant down regulation of VgR relative to the controls. GABA receptor also showed significant down-regulation between the EC 10 and EC 50 mixture treatments. Results of the present study demonstrate that pesticide-induced gene expression effects precede and occur at lower concentrations than organism-level responses. Integrating "omic" endpoints in traditional bioassays may thus be a promising way forward in pesticide toxicity evaluations. Graphical abstract Image 1 Highlights • The use of gene expression to unravel pesticide effects was evaluated. • Gene expression and traditional parameters after single and mixture exposure were compared. • Gene expression parameters complement "traditional" test parameters. • Gene expression analyses has great potential to increase TKTD understanding of pesticide stress. [ABSTRACT FROM AUTHOR]

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.

Conopomorpha sinensis Bradley is the dominant borer pest of Litchi chinesis and Euphoria longan. Current management of C. sinensis relies upon insecticide application to adult moths. In addition to the direct mortality induced by insecticides, a sublethal dose of insecticides also affects growth, survival, and reproduction in the exposed insects. Vitellogenin (Vg) and vitellogenin receptor (VgR) are normally identified as essential reproduction-related proteins in insects. In this study, we characterized these two genes from C. sinensis, and investigated their differential responses to sublethal concentrations of insecticide. Cloned CsVg and CsVgR consist of 5391 and 5424-bp open reading frames, which encode proteins of 1796 and 1807 amino acid residues, respectively. The CsVg protein contains the typical vitellogenin, DUF1943 and VWFD domains as other reported lepidopteran Vgs. The CsVgR was characterized as a typical low density lipoprotein receptor with two highly conserved LBD and EGF precursor domains, one hydrophobic transmembrane domain, one cytoplasmic domain, and 13 putative N-glycosylation sites. We next assessed the sublethal effect of four major insecticides on egg-laying in C. sinensis. The toxicity against C. sinensis varied among the insecticides tested, with LC50 values ranging from 0.23 ppm for chlorpyrifos to 20.00 ppm for β-cypermethrin, among which emamectin benzoate (EB) showed a significant negative impact on egg-laying, survival rate, ovarian development, and mating rate of C. sinensis at LC30 doses. Further investigation showed that the transcriptional level of CsVg and CsVgR were impaired in different way at 24, 48, and 72 h after EB exposure, and this result was in agreement with the diminished egg-laying of C. sinensis in the sublethal concentration EB-treated group. A repressed transcription level of CsVgR was observed at 48 h after treatment, suggesting that EB elicits a delayed response in the abundance of CsVgR. These results established different roles of CsVg and CsVgR in response to the sublethal effect of insecticides. CsVg might be a better parameter than CsVgR for assessing the effect of sublethal insecticides on reproduction in C. sinensis.

Conopomorpha sinensis Bradley is the dominant borer pest of Litchi chinesis and Euphoria longan. Current management of C. sinensis relies upon insecticide application to adult moths. In addition to the direct mortality induced by insecticides, a sublethal dose of insecticides also affects growth, survival, and reproduction in the exposed insects. Vitellogenin (Vg) and vitellogenin receptor (VgR) are normally identified as essential reproduction-related proteins in insects. In this study, we characterized these two genes from C. sinensis , and investigated their differential responses to sublethal concentrations of insecticide. Cloned CsVg and CsVgR consist of 5391 and 5424-bp open reading frames, which encode proteins of 1796 and 1807 amino acid residues, respectively. The CsVg protein contains the typical vitellogenin, DUF1943 and VWFD domains as other reported lepidopteran Vgs. The CsVgR was characterized as a typical low density lipoprotein receptor with two highly conserved LBD and EGF precursor domains, one hydrophobic transmembrane domain, one cytoplasmic domain, and 13 putative N-glycosylation sites. We next assessed the sublethal effect of four major insecticides on egg-laying in C. sinensis. The toxicity against C. sinensis varied among the insecticides tested, with LC50 values ranging from 0.23 ppm for chlorpyrifos to 20.00 ppm for β-cypermethrin, among which emamectin benzoate (EB) showed a significant negative impact on egg-laying, survival rate, ovarian development, and mating rate of C. sinensis at LC30 doses. Further investigation showed that the transcriptional level of CsVg and CsVgR were impaired in different way at 24, 48, and 72 h after EB exposure, and this result was in agreement with the diminished egg-laying of C. sinensis in the sublethal concentration EB-treated group. A repressed transcription level of CsVgR was observed at 48 h after treatment, suggesting that EB elicits a delayed response in the abundance of CsVgR. These results established different roles of CsVg and CsVgR in response to the sublethal effect of insecticides. CsVg might be a better parameter than CsVgR for assessing the effect of sublethal insecticides on reproduction in C. sinensis. [ABSTRACT FROM AUTHOR]

Abstract: Vitellogenin (Vg) and its receptor (VgR) play a key role in the reproductive process and development of insects. Aphids are a group of high‐fecundity insect species with pseudoplacental viviparity, but the roles of their Vg and VgR genes have not been investigated yet. The brown citrus aphid, Aphis (Toxoptera) citricidus, is a major insect pest of citrus and the main vector of Citrus tristeza closterovirus. In this study, we identified and characterized these two genes, designated as AcVg and AcVgR, from the brown citrus aphid. We found that AcVg has lost the DUF1943 domain that is present in other insect Vgs. Silencing of AcVg and AcVgR led to a delay in the nymph–adult transition, a prolonged prereproductive period, and a shortened reproductive period, which in turn resulted in slower embryonic development and fewer new‐born nymphs. Interestingly, silencing of AcVg decreased the transcript level of AcVgR, but silencing of AcVgR resulted in increased transcript levels of AcVg. In addition, silencing of Vg/VgR had similar phenotypes between alate and apterous morphs, suggesting that the functions of these two genes are the same in the two wing morphs of the aphid. Our results demonstrate that Vg and VgR are involved in various aspects of aphid development and reproduction. Further studies on the synthesis of Vg could help to elucidate the reproductive mechanism and provide information that will be useful for developing new pest control strategies. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: The beet armyworm Spodoptera exigua, a worldwide phytophagous pest, causes considerable economic agricultural losses. Understanding the relationship between its fecundity and the host plant is a basic and important component of early forecasting of beet armyworm outbreaks. However, little is known about the molecular mechanism by which distinct hosts affect S. exigua fecundity. RESULTS: In this study, key life‐history parameters of S. exigua reared on distinct hosts were investigated; the host plants could be ranked as lettuce > shallot > tomato > celery in their order of suitability. Full‐length S. exigua vitellogenin receptor (SeVgR) cDNA was cloned, and sex‐, stage‐ and tissue‐specific expression characteristics were assessed. Spodoptera exigua vitellogenin (SeVg) and SeVgR expression levels were markedly modulated by host nutrients (P < 0.05). SeVg and SeVgR expression levels were significantly higher in S. exigua reared on lettuce, the most preferred and most nutritive host, than in those reared on tomato and celery. Interestingly, significant linear regression correlations were found between SeVg and SeVgR expression levels and key S. exigua life‐history parameters, especially life span, pupa weight, and female fecundity (P < 0.01). CONCLUSION: Host plant type and suitability could affect the expression pattern of SeVg and SeVgR, which influenced S. exigua fecundity. Vg and VgR have the potential to be used as molecular markers of S. exigua fecundity and for forecasting outbreaks of S. exigua on different hosts. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Vitellogenin (Vg) and vitellogenin receptor (VgR) play important roles in the vitellogenesis of insects. In this study, we cloned and characterized the two corresponding genes (TpVg and TpVgR) in an economically important insect, Thitarodes pui (Lepidoptera: Hepialidae), from the Tibetan plateau. The full length of TpVg is 5566 bp with a 5373 bp open reading frame (ORF) encoding 1,790 amino acids. Sequence alignment revealed that TpVg has three conserved domains: a Vitellogenin_N domain, a DUF1943 domain, and a von Willebrand factor type D domain (VWD). The full length of TpVgR is 5732 bp, with a 5397 bp ORF encoding 1798 amino acids. BLASTP showed that TpVgR belongs to the low-density lipoprotein receptor (LDLR) gene superfamily. Structural analysis revealed that TpVgR has a group of four structural domains: a ligand-binding domain (LBD), an epidermal growth factor (EGF)-precursor homology domain, a transmembrane (TM) domain, and a cytoplasmic domain. In addition, TpVgR has four cysteine-rich LDL repeats in the first ligand-binding site and seven in the second. Quantitative real-time polymerase chain reaction analysis revealed that the expression levels of TpVg and TpVgR are much higher in later pupa than in either the larval or adult stage, implying that the synthesis and uptake of Vg in T. pui occurs in the later pupal stage. These results will help us to understand the molecular mechanism of the reproductive capacity and will provide new insight into the mass rearing and utilization of T. pui. [ABSTRACT FROM AUTHOR]

To evaluate potential involvement of clathrin in endocytosis of vitellogenin (Vtg) by teleost oocytes, cDNAs encoding clathrin heavy chain ( cltc ) were cloned from ovaries of cutthroat trout. Quantitative PCR revealed three types of cltc ( cltc-a1 , cltc-a2 , cltc-b ) to be expressed in 10 different tissues including the ovary. The cltc-a1 alone exhibited a significant decrease in ovarian expression during vitellogenesis; this was correlated with a corresponding decrease in transcripts encoding the major Vtg receptor (Vtgr). No development-related changes in ovarian cltc-a2 or cltc-b transcript levels were observed. In situ hybridization revealed a strong ctlc signal in pre-vitellogenic oocytes, but not in vitellogenic oocytes. Western blotting using a rabbit antiserum (a-Cltc) raised against a recombinant Cltc preparation detected a polypeptide band with an apparent mass of ~ 170 kDa in vitellogenic ovary extracts. Immunohistochemistry using a-Cltc revealed Cltc to be uniformly distributed throughout the ooplasm of perinucleolus stage oocytes, translocated to the periphery of lipid droplet stage oocytes, and localized to the oolemma during vitellogenesis. These patterns of cltc /Cltc distribution and abundance during oogenesis, which are identical to those previously reported for vtgr /Vtgr in this species, constitute the first empirical evidence that cltc-a1 /Cltc-a1 is involved in Vtg endocytosis via the Vtgr in teleost fish. [ABSTRACT FROM AUTHOR]

VgR, a member of the LDLR family, functions to transport vitellogenin into the ovaries to protome ovarian growth and embryonic development. In insects, the only widely accepted ligand of VgR is Vg. Recently, BmVgR has been shown to interact with BmSP1 in vitro. Therefore, in this study, we evaluated whether BmVgR could transport BmSP1 into certain cells. Although BmVgR could combine with BmVg and BmSP1, BmVgR did not affect the amount of BmSP1 taken up by Sf9 cells. Parallel immunofluorescence showed that most BmVg and BmVgR were localized in the inner oocyte membrane, showing tissue localization similar to that of BmVg labeled with pHrodo Red absorbed by the ovaries on day 2 of pupation. Although BmSP1 showed localization similar to BmVgR during the same phase, little BmSP1 was present in the ovary. Additionally, BmSP1 did not exist in ovaries when the ovaries contained BmVgR on day 5 of pupation, suggesting that BmSP1 in the ovaries was not endocytosed by BmVgR. In summary, BmVgR could facilitate uptake of BmVg by developing oocytes, but did not modulate in the transport of BmSP1. [ABSTRACT FROM AUTHOR]

Carmine spider mite, Tetranychus cinnabarinus (Boisduval), an agricultural pest of economically important crops, has developed resistance to a group of pesticides. We have selected a fenpropathrin-resistant strain (FeR) of T. cinnabarinus from the isogenous and susceptible strain (SS), and found that the FeR not only showed resistance but its fecundity also increased. According to the numbers of eggs laid per day of both strains, the FeR was more fertile than SS throughout the life span. To investigate the underlying reason, the protein contents of vitellogenin (vg) and vitellogenin receptor (vgr) were detected, and the results showed both of them were significantly higher in FeR than in SS. Then, the mRNA-expressions of vg and vgr genes were compared between FeR and SS. From the transcriptome data of T. cinnabarinus , we classified two vg genes (designated as Tcvg1 and Tcvg2 , respectively) and a vgr gene (designated as Tcvgr ). The expressions of Tcvg1 , Tcvg2 and Tcvgr were highly associated with the fecundity of the mites that their mRNAs were extremely abundant at the adult stage, but hardly detectable during the developmental period (from egg to deutonymph). In accordance with the protein content, the expression levels of the three genes were all significantly higher in FeR than they were in SS. These results suggested that after resistance selection with fenpropathrin in T. cinnabarinus , the fecundity and the expression of reproduction-related genes (vg and vgr) were significantly higher in fenpropathrin resistant strain than that in susceptible strain. [ABSTRACT FROM AUTHOR]

Chlordecone is a persistent organochlorine pesticide widely used in Guadeloupe (French West Indies) to control the banana weevil Cosmopolites sordidus. Although it was previously highlighted that chlordecone may affect the reproduction and growth of vertebrate species, little information is available on the chlordecone effects in invertebrates. The present study investigated the effects of chlordecone on a hormone and a protein having key roles in reproduction and growth of the decapod crustacean Macrobrachium rosenbergii, by measuring the 20-hydroxyecdysone concentration, vitellogenin, and vitellogenin receptor gene expression, as well as the bioconcentration of chlordecone in exposed prawns. First, the results revealed that chlordecone was accumulated in M. rosenbergii. Then, it was found that Vg and VgR gene expression were increased in male and female M. rosenbergii exposed to chlordecone for 90 and 240 days, while the 20-hydroxyecdysone concentrations were decreased. This work suggests that chlordecone accumulates in prawn tissues and could affect key molecules involved in the reproduction and the growth of the invertebrate M. rosenbergii. However, many questions remain unresolved regarding the impacts of chlordecone on growth and reproduction and the signaling pathways responsible for these effects, as well as the potential role of confounding factors present in in situ studies. [ABSTRACT FROM AUTHOR]

A complementary DNA (cDNA) that encodes the vitellogenin receptor (VgR) in the oriental river prawn, Macrobrachium nipponense , was cloned using expressed sequence tag analysis and a rapid amplification of cDNA ends approach. The coding region consists of 5920 base pairs (bp) that encode a 1902 amino acid protein, with a predicted molecular mass of 209 kDa. The coding region is flanked by a 45 bp 5ʹ-untranslated region (UTR) and a 166 bp 3ʹ-UTR. The deduced amino acid sequence of the M. nipponense VgR cDNA had typically conserved domains, such as an extracellular, lipoprotein-binding domain, epidermal growth factor-like and O -glycosylation domains, a transmembrane domain and a short C-terminal, cytosolic tail. Quantitative real-time PCR (qPCR) indicated that Mn-VgR is highly expressed in the female ovary. Expression analysis by qPCR demonstrated the larval and ovarian developmental stage-specific expression pattern. As the ovaries developed, the expression level of Mn-VgR gradually increased during the reproductive cycle (stage I), to reach a peak in stage III. Levels then dropped as a new development cycle was entered after reproduction molting. Eyestalk ablation led to a significant increase in the expression of Mn-VgR during the ovarian development stages ( P < 0.05), when compared with the eyestalk-intact group. The investigation revealed that eyestalk ablation initially affected Mn-VgR expression and then influenced vitellogenesis. In adult females, VgR RNA interference (RNAi) dramatically delayed the maturation of the ovary, in accordance with the gonad somatic index. In addition, Mn-VgR RNAi led to vitellin depletion in the oocytes and the accumulation of vitellin in the hepatopancreas. [ABSTRACT FROM AUTHOR]