RESUMO
Inappropriate application of pesticides not only causes sub-lethal effects on ecosystem service providers but also reduces crop yield and quality. As a xenobiotic signal molecule, pesticides may interact with signal transduction receptors in crops, resulting in oxidative damage and even metabolic perturbations. We discovered that three neonicotinoid insecticides (NIs), namely, imidacloprid, thiamethoxam, and clothianidin, at 0.06-0.12 kg ai/ha significantly inhibited the auxin signal pathway in rice leaves, thereby reducing the intracellular auxin (IAA) content. Molecular simulation further confirmed that NIs occupied the binding site where auxin transporter-like proteins 1 (LAX11) and 2 (LAX12), in which Thr253 and Asn66 of LAX11, as well as Thr244 and Asn57 of LAX12, were bound to the nitroguanidine of NIs via H-bonds. Meanwhile, Asn66 of LAX11 and Asn57 of LAX12 interacted with nitroguanidine via aromatic H-bonds. Moreover, phenylpropanoid biosynthesis was significantly disturbed because of the inhibited auxin signal pathway. Notably, peroxidase-coding genes were downregulated with a maximum value greater than 10-fold, resulting in decreased antioxidant metabolites flavone (37.82%) and lignin content (20.15%). Ultimately, rice biomass was reduced by up to 25.41% due to the decline in IAA content and antioxidant capacity. This study deeply explored the molecular mechanism of metabolic perturbations in crops stressed by pesticides, thus providing a scientific basis for pesticide environmental risk assessment and agricultural product safety.
Assuntos
Inseticidas , Neonicotinoides , Oryza , Antioxidantes/metabolismo , Ecossistema , Inseticidas/toxicidade , Neonicotinoides/toxicidade , Nitrocompostos/toxicidade , Transdução de SinaisRESUMO
Prostate cancer (PCa) is the most common malignant tumor in males, which frequently develops into castration-resistant prostate cancer (CRPC) with limited therapies. Gambogenic acid (GNA), a flavonoids compound isolated from Gamboge, exhibits anti-tumor capacity in various cancers. Our results showed that GNA revealed not only antiproliferative and pro-apoptotic activities but also the induction of autophagy in PCa cells. In addition, autophagy inhibitor chloroquine enhanced the pro-apoptosis effect of GNA. Moreover, the activation of JNK pathway and the induction of apoptosis and autophagy triggered by GNA were attenuated by JNK inhibitor SP600125. We also found that GNA significantly promoted reactive oxygen species (ROS) generation and endoplasmic reticulum (ER) stress. Meanwhile, suppressing ER stress with 4-phenylbutyric acid (4-PBA) markedly blocked the activation of JNK pathway induced by GNA. Further research indicated that ROS scavenger N-acetyl-L-cysteine (NAC) effectively abrogated ER stress and JNK pathway activation induced by GNA. Furthermore, NAC and 4-PBA significantly reversed GNA-triggered apoptosis and autophagy. Finally, GNA remarkably suppressed prostate tumor growth with low toxicity in vivo. In conclusion, the present study revealed that GNA induced apoptosis and autophagy through ROS-mediated ER stress via JNK signaling pathway in PCa cells. Thus, GNA might be a promising therapeutic drug against PCa.
Assuntos
Sistema de Sinalização das MAP Quinases , Neoplasias da Próstata , Masculino , Humanos , Espécies Reativas de Oxigênio/metabolismo , Apoptose , Estresse do Retículo Endoplasmático , Autofagia , Linhagem Celular Tumoral , Acetilcisteína/metabolismo , Acetilcisteína/farmacologia , Neoplasias da Próstata/tratamento farmacológicoRESUMO
Henosepilachna vigintioctopunctata is a serious defoliating beetle attacking Solanaceae and Cucurbitaceae plants in many Asian countries. In the present paper, we identified a putative myoglianin (myo) gene. Hvmyo was actively transcribed throughout development, from embryo to adult. RNA interference (RNAi)-aided knockdown of Hvmyo delayed larval development by more than 2 days, reduced larval body size, inhibited the growth of antennae, wings and legs and disturbed gut purge. Knockdown of Hvmyo impaired the larval-pupal transition. All the Hvmyo RNAi larvae arrested at the larval stage or formed misshapen pupae or adults. The deformed pupae and adults were partially wrapped with exuviae, bearing separated wings. Moreover, the expression levels of five ecdysteroidogenesis genes (Hvspo, Hvphm, Hvdib, Hvsad and Hvshd), a prothocicotropic hormone (PTTH)/Torso pathway gene (Hvtorso), two 20E receptor genes (HvEcR and HvUSP), and two 20E signalling genes (HvE93 and HvFTZ-F1) were as a result of HvMyo RNAi significantly lowered. Conversely, the expression of a JH biosynthesis gene (Hvjhamt), a JH receptor gene HvMet and a JH signalling gene HvKr-h1 was greatly enhanced. Although ingestion of 20E and Hal rescued the 20E signal, it could not alleviate larval performance and defective phenotypes. Our results suggest that Myo exerts four distinctive roles in ecdysteroidogenesis, JH production, organ growth and larva-pupa-adult transformation in H. vigintioctopunctata.
Assuntos
Besouros , Animais , Besouros/genética , Ecdisterona/metabolismo , Proteínas de Insetos/metabolismo , Metamorfose Biológica/genética , Pupa , Larva/genética , Interferência de RNA , Hormônios Juvenis/metabolismoRESUMO
Kynurenine pathway is critically important to catabolize tryptophan, to produce eye chromes, and to protect nervous system in insects. However, several issues related to tryptophan degradation remain to be clarified. In the present paper, we identified three genes (karmoisin, vermilion and cardinal) involved in kynurenine pathway in Henosepilachna vigintioctopunctata. The karmoisin and cardinal were highly expressed in the pupae and adults having compound eyes. Consistently, high-performance liquid chromatography result showed that three ommochrome peaks were present in adult heads rather than bodies (thoraces, legs, wings and abdomens). RNA interference (RNAi)-aided knockdown of vermilion caused accumulation of tryptophan in both adult heads and bodies, disappearance of ommochromes in the heads and a complete loss of eye color in both pupae and adults. Depletion of cardinal brought about excess of 3-hydroxykynurenine and insufficient ommochromes in the heads and decolored eyes. RNAi of karmoisin resulted in a decrease in ommochromes in the heads, and a partial loss of eye color. Moreover, a portion of karmoisin-, vermilion- or cardinal-silenced adults exhibited negative phototaxis, whereas control beetles showed positive phototaxis. Furthermore, dysfunctions of tryptophan catabolism impaired climbing ability. Our findings clearly illustrated several issues related to kynurenine pathway and provided a new insight into the physiological importance of tryptophan catabolism in H. vigintioctopunctata.
Assuntos
Vias Biossintéticas , Besouros/fisiologia , Proteínas de Insetos/metabolismo , Cinurenina/metabolismo , Larva/fisiologia , Fenotiazinas/metabolismo , Triptofano/metabolismo , Animais , Cor de Olho , Proteínas de Insetos/genética , LocomoçãoRESUMO
Insects rely heavily on their sophisticated chemosensory systems to locate host plants and find conspecific mates. Although the molecular mechanisms of odorant recognition in many Lepidoptera species have been well explored, limited information has been reported on the geometrid moth Ectropis obliqua Prout, an economically important pest of tea plants. In the current study, we first attempted to identify and characterize the putative olfactory carrier proteins, including odorant-binding proteins (OBPs) and chemosensory proteins (CSPs). By analyzing previously obtained transcriptomic data of third-instar larvae, five OBPs and 14 CSPs in E. obliqua were identified. Sequence alignment, conserved motif identification, and phylogenetic analysis suggested that candidate proteins have typical characteristics of the insect OBP or CSP family. The expression patterns regarding life stages and different tissues were determined by quantitative real-time PCR. The results revealed that four transcripts (OBP2, OBP4 and CSP8, CSP10) had larvae preferential expression profiles and nine candidate genes (PBP1, OBP1 and CSP2, CSP4, CSP5, CSP6, CSP7, CSP11, and CSP13) were adult-biased expressed. Further specific tissue expression profile evaluation showed that OBP1, OBP2, OBP4, and PBP1 were highly expressed at olfactory organs, implying their potential involvement in chemical cue detection, whereas CSPs were ubiquitously detected among all of the tested tissues and could be associated with multiple physiological functions. This study provided a foundation for understanding the physiological functions of OBPs and CSPs in E. obliqua and will help pave the way for the development of a new environmental friendly pest management strategy against the tea geometrid moth.
Assuntos
Proteínas de Insetos/genética , Mariposas/genética , Receptores Odorantes/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Proteínas de Insetos/química , Larva , Masculino , Filogenia , Receptores Odorantes/química , Alinhamento de Sequência , Olfato , TranscriptomaRESUMO
A new class of fluorescein/rhodamine hybrids with two spirolactone rings was reported to exhibit dual-output fluorescent behaviors independently. Isolation and characterization for two diastereomers, trans-RhOH and cis-RhOH, have been made and their X-ray crystal structures determined. In a basic environment, the spirolactone ring on the hydroxyl side will be opened to give a fluorescein-like optical output with the lowest absorptions at 485 and 530 nm emission. On the other hand, a rhodamine-like optical output, i.e., 528 nm absorption and 575 nm emission, will be switched on by a H(+) or a Hg(2+) ion, attributed to the spirolactone ring opening on the amino side. In a methanol-buffer system with different pH values, the corresponding pKa values for the hydroxyl and amino groups were determined as 5.7 and 2.3, respectively. Selective Hg(2+)-sensing properties have also been discussed, and log Ks values of about 3.60 and 3.73 were determined. Confocal microscopic images of Caenorhabditis elegans incubated with RhOH were found to show enhanced fluorescent intensity with a Hg(2+) ion, demonstrating the potential application of RhOH for in vivo biological imaging.
Assuntos
Fluoresceína/química , Rodaminas/química , Animais , Caenorhabditis elegans/efeitos dos fármacos , Cristalografia por Raios X , Fluoresceína/farmacologia , Rodaminas/farmacologiaRESUMO
Plant tolerance to abiotic stress depends on fast molecular cascades involving stress perception, signal transduction, gene expression alterations, and metabolic rearrangement. This study sheds light on the tolerance mechanism of rice (Oryza sativa L.) towards the toxicity of the polycyclic aromatic hydrocarbons (PAHs), including phenanthrene (Phe), pyrene (Pyr), and benzo[a]pyrene (BaP). Results showed that three PAHs significantly activated the phosphoinositide signaling system involving the phosphorus (P) metabolism and homeostasis in rice roots. This activation increased phytic acid (IP6) levels to over 54.12% of the control (p < 0.05). Molecular docking verified that three PAHs occupied the IP6 binding site in SPX3, a negative regulatory factor of P homeostasis, where ARG229 interacted with PAHs via the van der Waals force. Moreover, the expression of gene encoding SPX3 was significantly downregulated 2.81-, 2.83-, and 2.18-fold under Phe, Pyr, and BaP stress, respectively, relative to the control. Conversely, the expression levels of the gene coding SDEL2 was significantly increased, promoting the degradation of SPX3. Ultimately, P absorption and nucleic acid synthesis were enhanced, alleviating the inhibition effect of PAHs on rice growth. Notably, Pyr demonstrated the strongest binding affinity for SPX3, confirming its critical interference with P homeostasis. These findings provide insight into the molecular mechanisms regulating plant responses to PAHs, and offer guidance for improving crop resistance against organic pollutants and protecting food security.
Assuntos
Oryza , Hidrocarbonetos Policíclicos Aromáticos , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Oryza/genética , Oryza/metabolismo , Simulação de Acoplamento Molecular , Homeostase , Transdução de SinaisRESUMO
Polycyclic aromatic hydrocarbons (PAHs) are potent inhibitors of DNA that can induce genetic damage, abnormal gene expression, and metabolic disorders upon interfacing with biological macromolecules. However, the mechanism of their interactions with DNA remains elusive. Therefore, this study selected three representative PAHs, including phenanthrene (Phen), pyrene (Pyre), and benzo[a]pyrene (B[a]P), and explored their binding mechanisms with the double-strand DNA (dsDNA) from different species, including 1J1V (Escherichia coli), 6J5B (Arabidopsis thaliana), and 6Q1V (Homo sapiens). The results revealed that binding between PAHs and dsDNA occurred in the groove via van der Waals forces and π-π stacking, with the carboxyl oxygen atom of the thymine (T)-base within dsDNA being the key binding site. This result was further confirmed by the spectroscopic experiments, where significant changes in the peak of the T-base were observed after PAHs-dsDNA binding. More interestingly, the total binding energies of Pyre with the three dsDNA were -138.800 kJ/mol (Pyre-1J1V), -105.523 kJ/mol (Pyre-6J5B), and -127.567 kJ/mol (Pyre-6Q1V), respectively, all of which were higher than those of Phen and B[a]P. This suggests that that Pyre has the strongest dsDNA binding ability. Additionally, analysis of the thermodynamic parameters indicated that the interactions between the three PAHs and dsDNA were exothermic reactions. In contrast, the Pyre-dsDNA interaction predominantly involved van der Waals forces and hydrogen bonding due to the enthalpy change (∆H) < 0 and entropy change (∆S) < 0, while the Phen-dsDNA and B[a]P-dsDNA interactions predominantly involved hydrophobic forces due to ∆H > 0 and ∆S > 0. Furthermore, Pyre caused local distortion of dsDNA, which was more pronounced under atomic force microscopy (AFM). In summary, this study has unveiled a new phenomenon of binding between PAHs and dsDNA. This sheds light on the carcinogenic potential and environmental impacts of PAHs pollution.
Assuntos
DNA , Hidrocarbonetos Policíclicos Aromáticos , Timina , Timina/química , Humanos , Arabidopsis/metabolismo , Escherichia coli , Fenantrenos , Pirenos/químicaRESUMO
Tricyclic antidepressant is an old and well-established therapeutic agent with a good safety profile, making them an excellent candidate for repurposing. In light of the growing understanding of the importance of nerves in the development and progression of cancer, attention is now being turned to using nerve-targeting drugs for the treatment of cancer, particularly TCAs. However, the specific mechanism by which antidepressants affect the tumor microenvironment of glioblastoma (GBM) is still unclear. Here, we combined bulk RNA sequencing, network pharmacology, single-cell sequencing, molecular docking and molecular dynamics simulation to explore the potential molecular mechanism of imipramine in the treatment of GBM. We first revealed that the imipramine treatment is presumed to target EGFRvIII and neuronal-derived EGFR, which may play a pivotal role in treating GBM by reducing the GABAergic synapse and vesicle-mediated release and other processes thereby modulating immune function. The novel pharmacological mechanisms might provide further research directions.
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Tetrahydrobiopterin (BH4) is produced from guanosine triphosphate (GTP) under catalyzation of GTP cyclohydrolase I (GTPCH), 6-pyruvoyltetrahydropterin synthase (PTPS) and sepiapterin reductase (SR), among others. In Drosophila melanogaster, BH4 and other pteridines are required for cuticle tanning and eye pigmentation. In this study, two Hvgtpch (Hvgtpch-a and Hvgtpch-b), an Hvptps and an Hvsr transcripts were identified in a serious defoliator Henosepilachna vigintioctopunctata. Hvgtpch-a and Hvgtpch-b were highly expressed just before and/or right after the molt, in contrast to Hvptps and Hvsr. RNA interference (RNAi) by injection of a dsgtpch targeting the common fragment of Hvgtpch-a and Hvgtpch-b into the third instar larvae caused albino fourth-instar larvae and pupae. Around 80% of the Hvgtpch RNAi larvae failed to pupate. The remaining 20% of Hvgtpch RNAi pupated beetles did not completely remove the larval/pupal exuviae after emerged as adults and eventually died. Depletion of Hvgtpch at the fourth instar stage resulted in under-pigmented pupae and adults, with significantly low pupation and emergence rates. The Hvgtpch RNAi adults rarely moved and fed on plant leaves; they died within a week after emergence. Silence of Hvptps or Hvsr at the third- and fourth-instar stages led to similar but less serious phenotypes, with lowest influence in the Hvsr RNAi ladybirds. Moreover, RNAi of Hvgtpch, Hvptps or Hvsr did not affect coloration of the larval ocelli and pupal/adult compound eyes. Therefore, our results demonstrated that pteridines are involved in melanin formation but not in eye pigmentation in H. vigintioctopunctata. Moreover, our findings will enable the development of a dsgtpch-based pesticide to control H. vigintioctopunctata larvae.
Assuntos
Besouros , Drosophila melanogaster , Animais , Besouros/metabolismo , Larva , Biopterinas/metabolismo , Pigmentação , PupaRESUMO
Bladder cancer is one of the most common carcinomas in the human urinary system worldwide. Loperamide, known as an antidiarrheal drug, exerts anti-tumor activities against various cancers. However, the effect of loperamide on bladder cancer cells remains unclear. Our study aimed to investigate the effect of loperamide on bladder cancer and explore the underlying mechanisms. We found that loperamide suppressed the proliferation of 5637 and T24 cells in a dose-dependent manner. Loperamide treatment showed both pro-apoptotic and pro-autophagic effects on bladder cancer cells. Moreover, it was revealed that loperamide induced reactive oxygen species (ROS) accumulation, leading to the activation of c-Jun N-terminal kinase (JNK) signaling pathway. Notably, ROS scavenger N-acetyl-L-cysteine (NAC) and JNK inhibitor SP600125 effectively attenuated the induction of autophagy and apoptosis triggered by loperamide. Finally, blocking autophagy with CQ could significantly enhance the anti-cancer effect of loperamide both in vitro and in vivo. Overall, these findings demonstrated that loperamide induced autophagy and apoptosis through the ROS-mediated JNK pathway in bladder cancer cells. Our results suggest that the strategy of combining loperamide with autophagy inhibitor CQ may provide a therapeutic option for the treatment of bladder cancer.
Assuntos
Sistema de Sinalização das MAP Quinases , Neoplasias da Bexiga Urinária , Humanos , Loperamida/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Linhagem Celular Tumoral , Apoptose , Neoplasias da Bexiga Urinária/tratamento farmacológico , Autofagia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismoRESUMO
Insect development is regulated by a combination of juvenile hormone (JH) and 20-hydroxyecdysone (20E). Production of both JH and 20E is regulated by transforming growth factor-ß (TGFß) signaling. TGFß can be classified into two branches, the Activin and Bone Morphogenetic Protein (BMP) pathways. In Drosophila melanogaster, BMP signaling is critical for JH synthesis, whereas Activin signal is required to generate the large pulse of 20E necessary for entering metamorphosis. However, to which extent the roles of these signals are conserved remains unknown. Here we studied the role of an Activin component Smad onâ¯Xâ¯(Smox) in post-embryonic development in a defoliating ladybird Henosepilachna vigintioctopunctata. RNA interference (RNAi)-aided knockdown of Hvsmox inhibited larval growth, and impaired larval development. All Hvmyo RNAi larvae arrested at the fourth-instar larval stage. Moreover, knockdown of Hvsmox delayed gut and Malpighian tubules remodeling. Furthermore, the expression of a JH biosynthesis gene (Hvjhamt), a JH receptor gene HvMet and a JH response gene HvKr-h1 was greatly enhanced. Conversely, the expression levels of an ecdysteroidogenesis gene (Hvspo), a 20E receptor gene (HvEcR) and six 20E response genes (HvBrC, HvE74, HvE75, HvE93, HvHR3 and HvHR4) were significantly lowered. Knockdown of HvMet partially restored the negative phenotypes in the Hvsmox RNAi beetles. Our results suggest that Smox exerts regulative roles in JH production, ecdysteroidogenesis and organ remodeling, thus contributing to modulate the larva-pupa-adult transformation in H. vigintioctopunctata.
Assuntos
Besouros , Ativinas/genética , Ativinas/metabolismo , Animais , Besouros/metabolismo , Drosophila melanogaster/genética , Ecdisterona/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/metabolismo , Hormônios Juvenis/metabolismo , Larva , Metamorfose Biológica/genética , Pupa , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
Ecdysone-induced protein 93F (E93) plays triple roles during post-embryonic development in insects whose juvenile instars are more than four. However, it only acts as a specifier of adult structures in Drosophila flies whose larval instars are fixed at three. In this study, we determined the functions of E93 in the eggplant lady beetle (Henosepilachna vigintioctopunctata), which has four larval instars. We uncovered that E93 was abundantly expressed at the prepupal and pupal stages. A precocious inhibition of the juvenile hormone signal by RNA interference (RNAi) of HvKr-h1 or HvHairy, two vital downstream developmental effectors, at the penultimate instar larval stage increased the expression of E93, Conversely, ingestion of JH by the third-instar larvae stimulated the expression of HvKr-h1 but repressed the transcription of either HvE93X1 or HvE93X2. However, disturbance of the JH signal neither drove premature metamorphosis nor caused supernumerary instars. In contrast, depletion of E93 at the third- and fourth-instar larval and prepupal stages severely impaired pupation and caused a larval-pupal mixed phenotype: pupal spines and larval scoli were simultaneously presented on the cuticle. RNAi of E93 at the pupal stage affected adult eclosion. When the beetles had suffered from a dsE93 injection at the fourth-instar larval and pupal stages, a few resultant adults emerged, with separated elytra, abnormally folded hindwings, a small body size and short appendages. Taken together, our results suggest the larval instars are fixed in H. vigintioctopunctata; E93 serves as a repressor of larval characters and a specifier of adult structures during the larval-pupal-adult transition.
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Activation of inflammation is an important pathogenic factor contributing to the development of chronic kidney disease (CKD). Recent studies manifested the implication of impaired mitophagy mediated NLRP3 inflammasome activation in the progression of CKD. Mitochondria-targeted antioxidant mitoTEMPO showed antioxidant and anti-inflammatory properties in kidney disease. This study aims to investigate the protective mechanism of mitoTEMPO on podocyte injury related to mitophagy and NLRP3 inflammasome. Our results showed that mitoTEMPO obviously ameliorated renal function and podocyte injury in CKD model rats induced by cationic bovine serum albumin (C-BSA). More importantly, mitoTMEPO significantly inhibited NLRP3 inflammasome activation compared with CKD model rats (P < 0.01). In vitro, TNF-α damaged human podocyte cells (HPC) and activated NLRP3 inflammasome, which was rescued by NLRP3 inhibitor and mitoTEMPO. Meanwhile, mitoTEMPO lessened excessive mitochondrial ROS (mtROS) and degressive mitochondrial membrane potential (MMP) in HPC. We also found that mitoTEMPO induced mitophagy in vivo and in vitro. Moreover, silenced Parkin dramatically reserved the inhibitory effect of mitoTEMPO on NLRP3 inflammasome. Taking together, these findings reveal that mitoTEMPO ameliorated podocyte injury by inhibiting NLRP3 inflammasome via PINK1/Parkin pathway-mediated mitophagy. MitoTEMPO may be a new candidate to protect against podocyte injury in CKD.
Assuntos
Podócitos , Insuficiência Renal Crônica , Animais , Antioxidantes/farmacologia , Humanos , Inflamassomos , Mitofagia , Proteína 3 que Contém Domínio de Pirina da Família NLR , Compostos Organofosforados , Piperidinas , Proteínas Quinases , Ratos , Espécies Reativas de Oxigênio , Ubiquitina-Proteína LigasesRESUMO
Fushi Tarazu Factor 1 (FTZ-F1), a member of the nuclear receptor superfamily, is the downstream factor of 20-hydroxyecdysone signaling. In Drosophila melanogaster, alternative transcription start and splicing in the FTZ-F1 gene generate αFTZ-F1 and ßFTZ-F1 isoforms, which are vital for pair-rule segmentation in early embryogenesis and post-embryonic development, respectively. However, whether the same mRNA isoforms are present and exert the conservative roles remains to be clarified in other insects. In the present paper, we first mined the genomic data of representative insect species and unveiled that the same post-transcriptional processing in FTZ-F1 occurred in coleopterans, lepidopterans, dipterans and hymenopterans. Our expression data in Henosepilachna vigintioctopunctata, a serious polyphagous defoliator damaging a wide range of crops in Solanaceae and Cucurbitaceae, showed that both αFTZ-F1 and ßFTZ-F1 were actively transcribed throughout the development, from embryo to adult. The RNA interference-aided knockdown of both isoforms completely arrested larval ecdysis from the third to the fourth instar, in contrast to the depletion of either isoform. In contrast, silencing ßFTZ-F1, rather than αFTZ-F1, severely impaired the larval-pupal transformation. We accordingly propose that both FTZ-F1 isoforms are essential but mutually interchangeable for larval-larval molting, while ßFTZ-F1 is necessary for the larval-pupal transition and sufficient to exert the role of both FTZ-F1s during larval-pupal metamorphosis in H. vigintioctopunctata.
RESUMO
Henosepilachna vigintioctopunctata is a serious insect pest which attacks a large number of nightshades and cucurbits in Asian countries, Brazil and Australia. Prolonged application of traditional pesticides has caused environmental pollution and exerted deleterious effects on human health. Finding new approaches with high target specificity and low environmental contamination has become an urgent task. RNA interference (RNAi) induced by double-stranded RNA (dsRNA) is expected to be applicable to managing this pest. Here we evaluated the effects of Escherichia coli-expressed dsRNAs targeting ecdysone receptor (EcR) gene via dietary delivery in laboratory and foliar spraying in a greenhouse. The target transcript was successfully knocked down when the 4th-instar larvae had fed on potato foliage dipped with dsEcR in a laboratory bioassay. Around 85% of the HvEcR RNAi larvae remained as prepupae or became abnormal pupae, and failed to emerge into adults. Ingestion of dsEcR-immersed foliage by the 3rd-instar larvae effectuated a comparable RNAi response and brought about more severe defects: all the resultant larvae arrested development, remained as prepupae and finally died. For assay in the greenhouse, a dsEcR-contained E. coli suspension was directly sprayed to the foliage of greenhouse-growing potato plants and the 3rd- and 4th-instar larvae were transferred to the leaves. High RNAi efficacy was obtained and identical RNAi phenotypes were observed in treated larvae. In addition, spraying dsEcR reduced leaf damage. Our results indicate a possibility of practical application of dsEcR as an environmentally friendly RNA pesticide to control H. vigintioctopunctata larvae.
Assuntos
Besouros/crescimento & desenvolvimento , Proteínas de Insetos/genética , Interferência de RNA , Receptores de Esteroides/genética , Animais , Besouros/genética , Besouros/metabolismo , Escherichia coli , Proteínas de Insetos/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Microrganismos Geneticamente Modificados , Pupa/genética , Pupa/crescimento & desenvolvimento , Pupa/metabolismo , RNA de Cadeia Dupla/genética , RNA de Cadeia Dupla/metabolismo , Receptores de Esteroides/metabolismoRESUMO
According to the characteristics of the textile fibers Raman spectra, a qualitative identification method based on Raman feature extraction is proposed. This fast method consists of spectrum measurement and spectral data processing algorithm, including spectrum preprocessing, feature extraction and matching recognition. It can be used to identify the components of fibers or fabrics, especially chemical fibers, which is an inspective difficulty in daily analytic work for its remarkable Raman feature. The authors performed an experiment to analyze 4 typical and widely used kinds of fibers as algorithm verification. They are terylene fiber, acrylic fiber, nylon fiber and rayon fiber. To identify the components of one test sample, first the authors set up feature tables of these 4 standard samples, which describe the features of their preprocessed spectra containing both position information and intensity information, then extract features of the test sample. The authors match these features with the tables and calculate the matching confidence coefficients of the results, which can be used to filter the unexpected matching results caused by accident and attain the final qualitative identification result. The experimental results confirm that this method is effective, efficient and expansible, which means it can be used to identify more actual fiber types by adding more standard spectra to the feature table database. In addition, it is a pure optical method, which needs only a small quantity of sample without any pretreatment. The whole identification process is damage-free, pollution-free and suitable for various kinds of fabrics. Compared to all existing methods, this Raman spectrum identification method can solve the limitation of efficiency, pollution, universality, and fill a gap in fabric inspection field.
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Cell division requires constriction of an actomyosin ring to segregate the genetic material equally into two daughter cells. The spatial and temporal regulation of the contractile ring at the division plane primarily depends on intracellular signals mediated by the centralspindlin complex and astral microtubules. Although much investigative work has elucidated intracellular factors and mechanisms controlling this process, the extracellular regulation of cytokinesis remains unclear. Thus far, the extracellular matrix protein Hemicentin (HIM-4) has been proposed to be required for cleavage furrow stabilization. The underlying molecular mechanism, however, has remained largely unknown. Here, we show that HIM-4 and anillin (ANI-1) genetically act in the same pathway to maintain the rachis bridge stability in the germline. Our FRAP experiments further reveal that HIM-4 restricts the motility of ANI-1. In addition, we demonstrate that HIM-4 is recruited to the cleavage site in dividing germ cells and promotes the proper ingression of the cleavage membrane. Collectively, we propose that HIM-4 is an extracellular factor that regulates ANI-1 for germ cell membrane stabilization and contractile ring formation in Caenorhabditis elegans germline cells.
Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/citologia , Proteínas Contráteis/metabolismo , Citocinese/fisiologia , Proteínas da Matriz Extracelular/metabolismo , Células Germinativas/metabolismo , Proteínas de Membrana/metabolismo , Proteínas dos Microfilamentos/metabolismo , Animais , Proteínas de Caenorhabditis elegans/genética , Membrana Celular/metabolismo , Segregação de Cromossomos/fisiologia , Escherichia coli/genética , Técnicas de Introdução de Genes , Proteínas dos Microfilamentos/genética , Interferência de RNARESUMO
Larvae of the tapeworm Echinococcus multilocularis cause alveolar echinococcosis (AE), one of the most lethal helminthic infections in humans. The germinative cells, a population of stem cell-like cells, are considered to drive the continuous growth of the metacestodes within the host. The mechanisms and relative molecules controlling the behavior of germinative cells are poorly understood. Sox transcription factors play important roles in maintenance and regulation of stem/progenitor cells. We here describe the identification of a Sox family member in E. multilocularis, EmSOX2, as a potential regulator of germinative cells. Replacement of mouse Sox2 with EmSox2 could derive induced pluripotent stem cells (iPSCs) from somatic cells, suggesting that EmSOX2 is functionally related to mammalian SOX2. EmSOX2 is actively expressed in the proliferating germinative cells in E. multilocularis, and is significantly downregulated upon specific depletion of the germinative cell population by hydroxyurea treatment. These findings suggest that EmSOX2 may play a critical role in regulating E. multilocularis germinative cells.
Assuntos
Echinococcus multilocularis/metabolismo , Regulação da Expressão Gênica/fisiologia , Fatores de Transcrição SOXB1/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , Marcadores Genéticos , Células HEK293 , Humanos , Reprodução , Fatores de Transcrição SOXB1/genéticaRESUMO
BACKGROUND: Larvae of the tapeworm E. multilocularis cause alveolar echinococcosis (AE), one of the most lethal helminthic infections in humans. A population of stem cell-like cells, the germinative cells, is considered to drive the larval growth and development within the host. The molecular mechanisms controlling the behavior of germinative cells are largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: Using in vitro cultivation systems we show here that the EGFR/ERK signaling in the parasite can promote germinative cell proliferation in response to addition of human EGF, resulting in stimulated growth and development of the metacestode larvae. Inhibition of the signaling by either the EGFR inhibitors CI-1033 and BIBW2992 or the MEK/ERK inhibitor U0126 impairs germinative cell proliferation and larval growth. CONCLUSIONS/SIGNIFICANCE: These data demonstrate the contribution of EGF-mediated EGFR/ERK signaling to the regulation of germinative cells in E. multilocularis, and suggest the EGFR/ERK signaling as a potential therapeutic target for AE and perhaps other human cestodiasis.