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1.
BMC Genomics ; 22(1): 639, 2021 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-34479486

RESUMO

BACKGROUND: Resistance of pest insect species to insecticides, including B. thuringiensis (Bt) pesticidal proteins expressed by transgenic plants, is a threat to global food security. Despite the western corn rootworm, Diabrotica virgifera virgifera, being a major pest of maize and having populations showing increasing levels of resistance to hybrids expressing Bt pesticidal proteins, the cell mechanisms leading to mortality are not fully understood. RESULTS: Twenty unique RNA-seq libraries from the Bt susceptible D. v. virgifera inbred line Ped12, representing all growth stages and a range of different adult and larval exposures, were assembled into a reference transcriptome. Ten-day exposures of Ped12 larvae to transgenic Bt Cry3Bb1 and Gpp34/Tpp35Ab1 maize roots showed significant differential expression of 1055 and 1374 transcripts, respectively, compared to cohorts on non-Bt maize. Among these, 696 were differentially expressed in both Cry3Bb1 and Gpp34/Tpp35Ab1 maize exposures. Differentially-expressed transcripts encoded protein domains putatively involved in detoxification, metabolism, binding, and transport, were, in part, shared among transcripts that changed significantly following exposures to the entomopathogens Heterorhabditis bacteriophora and Metarhizium anisopliae. Differentially expressed transcripts in common between Bt and entomopathogen treatments encode proteins in general stress response pathways, including putative Bt binding receptors from the ATP binding cassette transporter superfamily. Putative caspases, pro- and anti-apoptotic factors, as well as endoplasmic reticulum (ER) stress-response factors were identified among transcripts uniquely up-regulated following exposure to either Bt protein. CONCLUSIONS: Our study suggests that the up-regulation of genes involved in ER stress management and apoptotic progression may be important in determining cell fate following exposure of susceptible D. v. virgifera larvae to Bt maize roots. This study provides novel insights into insect response to Bt intoxication, and a possible framework for future investigations of resistance mechanisms.


Assuntos
Bacillus thuringiensis , Besouros , Praguicidas , Animais , Bacillus thuringiensis/genética , Sobrevivência Celular , Besouros/genética , Endotoxinas/toxicidade , Resistência a Inseticidas , Larva/genética , Controle Biológico de Vetores , Plantas Geneticamente Modificadas/genética , Regulação para Cima , Zea mays/genética
2.
J Hazard Mater ; 416: 126142, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34492931

RESUMO

2-Methylisoborneol (2-MIB), a natural odorous substance, is widely distributed in water environment, but there is a paucity of information concerning its systemic toxicity. Herein, we investigated the effects of 2-MIB exposure on developmental parameters, locomotive behavior, oxidative stress, apoptosis and transcriptome of zebrafish. Zebrafish embryos exposed to different concentrations (0, 0.5, 5 and 42.8 µg/L) of 2-MIB showed no changes in mortality, hatchability, and malformation rate, but the body length of zebrafish larvae was significantly increased in a dose-dependent manner, and accompanied by the changes of growth hormone/insulin-like growth factor (GH/IGF) axis and the hypothalamic-pituitary-thyroid (HPT) axis genes. Moreover, the swimming activity of zebrafish larvae increased, which may be due to the increase of acetylcholinesterase (AChE) activity. Meanwhile, 2-MIB caused oxidative stress and apoptosis in zebrafish larvae by altering the NF-E2-related factor 2 (Nrf2) and mitochondrial signaling pathways, respectively. Transcriptome sequencing assay showed that the phototransduction signaling pathway was significantly enriched, and most of the genes in this pathway exhibited enhanced expression after exposure to 2-MIB. These findings provide an important reference for risk assessment and early warning to 2-MIB exposure.


Assuntos
Poluentes Químicos da Água , Peixe-Zebra , Animais , Embrião não Mamífero , Larva/genética , Transcriptoma , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/genética
3.
Pestic Biochem Physiol ; 178: 104946, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34446184

RESUMO

Spodoptera litura (Fabricius) is a widely distributed, highly polyphagous pest that can cause severe damage to a variety of economically important crops. Various populations have developed resistance to different classes of insecticides. In this study, we report on two indoxacarb-resistant S. litura populations, namely Ind-R (resistance ratio = 18.37-fold) derived from an indoxacarb-susceptible (Ind-S) population and a population caught from a field (resistance ratio = 46.72-fold). A synergist experiment showed that piperonyl butoxide (PBO) combined with indoxacarb produced higher synergistic effects (synergist ratio = 5.29) in the Ind-R population as compared to Ind-S (synergist ratio = 3.08). Elevated enzyme activity of cytochrome P450 monooxygenases (P450s) was observed for Ind-R (2.15-fold) and the Field-caught population (4.03-fold) as compared to Ind-S, while only minor differences were noticed in the activities of esterases and glutathione S-transferases. Furthermore, expression levels of P450 genes of S. litura were determined by quantitative reverse transcription PCR to explore differences among the three populations. The results showed that the mRNA levels of CYP6AE68, a novel P450 gene belonging to the CYP6 family, were constitutively overexpressed in Ind-R (32.79-fold) and in the Field-caught population (68.11-fold). CYP6AE68 expression in S. litura was further analyzed for different developmental stages and in different tissues. Finally, we report that RNA interference-mediated silencing of CYP6AE68 increased the mortality of fourth-instar larvae exposed to indoxacarb at the LC50 dose level (increase by 33.89%, 29.44% and 22.78% for Ind-S, Ind-R and the Field-caught population, respectively). In conclusion, the findings of this study indicate that expression levels of CYP6AE68 in S. litura larvae are associated with indoxacarb resistance and that CYP6AE68 may play a significant role in detoxification of indoxacarb.


Assuntos
Inseticidas , Mariposas , Animais , Sistema Enzimático do Citocromo P-450/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Larva/genética , Oxazinas/farmacologia , Spodoptera/genética
4.
J Exp Biol ; 224(7)2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-34424982

RESUMO

The ABC transporter ABCB1 plays an important role in the disposition of xenobiotics. Embryos of most species express high levels of this transporter in early development as a protective mechanism, but its native substrates are not known. Here, we used larvae of the sea urchin Strongylocentrotus purpuratus to characterize the early life expression and role of Sp-ABCB1a, a homolog of ABCB1. The results indicate that while Sp-ABCB1a is initially expressed ubiquitously, it becomes enriched in the developing gut. Using optimized CRISPR/Cas9 gene editing methods to achieve high editing efficiency in the F0 generation, we generated ABCB1a crispant embryos with significantly reduced transporter efflux activity. When infected with the opportunistic pathogen Vibrio diazotrophicus, Sp-ABCB1a crispant larvae demonstrated significantly stronger gut inflammation, immunocyte migration and cytokine Sp-IL-17 induction, as compared with infected control larvae. The results suggest an ancestral function of ABCB1 in host-microbial interactions, with implications for the survival of invertebrate larvae in the marine microbial environment.


Assuntos
Sistemas CRISPR-Cas , Ouriços-do-Mar , Animais , Sistemas CRISPR-Cas/genética , Imunidade , Larva/genética , Mutagênese , Vibrio
5.
BMC Genomics ; 22(1): 584, 2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34340656

RESUMO

BACKGROUND: Animal behavior is largely driven by the information that animals are able to extract and process from their environment. However, the function and organization of sensory systems often change throughout ontogeny, particularly in animals that undergo indirect development. As an initial step toward investigating these ontogenetic changes at the molecular level, we characterized the sensory gene repertoire and examined the expression profiles of genes linked to vision and chemosensation in two life stages of an insect that goes through metamorphosis, the butterfly Bicyclus anynana. RESULTS: Using RNA-seq, we compared gene expression in the heads of late fifth instar larvae and newly eclosed adults that were reared under identical conditions. Over 50 % of all expressed genes were differentially expressed between the two developmental stages, with 4,036 genes upregulated in larval heads and 4,348 genes upregulated in adult heads. In larvae, upregulated vision-related genes were biased toward those involved with eye development, while phototransduction genes dominated the vision genes that were upregulated in adults. Moreover, the majority of the chemosensory genes we identified in the B. anynana genome were differentially expressed between larvae and adults, several of which share homology with genes linked to pheromone detection, host plant recognition, and foraging in other species of Lepidoptera. CONCLUSIONS: These results revealed promising candidates for furthering our understanding of sensory processing and behavior in the disparate developmental stages of butterflies and other animals that undergo metamorphosis.


Assuntos
Borboletas , Animais , Borboletas/genética , Feminino , Expressão Gênica , Perfilação da Expressão Gênica , Genoma , Larva/genética , Masculino , Transcriptoma
6.
Braz J Biol ; 83: e245202, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34378662

RESUMO

Although propolis has been reported for having anti-inflammatory activities, its effects on complement system has not been much studied. This research was conducted to find out the effects of Indonesian propolis on the expression levels of C3, C1r/s, Bf, MBL, and C6 in zebrafish larvae which were induced by lipopolysaccharide (LPS). Counting of macrophages migrating to yolk sac and liver histology were carried out. Larvae were divided into four groups: CON (cultured in E3 medium only), LPS (cultured in a medium containing 0.5 µg/L LPS), LPSIBU (cultured in a medium containing LPS, and then treated with 100 µg/L ibuprofen for 24 hours), and LPSPRO (cultured in a medium containing LPS, and then immersed in 14,000 µg/L propolis for 24 hours) groups. The results showed that complement gene expression in larvae from the LPSIBU and LPSPRO groups were generally lower than in larvae from the LPS group. The number of macrophage migrations to the yolk in the LPSPRO group was also lower than in the LPS group. Histological structure of liver in all groups were considered normal. This study shows that Indonesian propolis has the potential to be used as an alternative to the substitution of NSAIDs.


Assuntos
Própole , Peixe-Zebra , Animais , Regulação para Baixo , Indonésia , Larva/genética , Lipopolissacarídeos/farmacologia , Própole/farmacologia , Peixe-Zebra/genética
7.
Int J Mol Sci ; 22(15)2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34361081

RESUMO

Cancer cachexia is a common deleterious paraneoplastic syndrome that represents an area of unmet clinical need, partly due to its poorly understood aetiology and complex multifactorial nature. We have interrogated multiple genetically defined larval Drosophila models of tumourigenesis against key features of human cancer cachexia. Our results indicate that cachectic tissue wasting is dependent on the genetic characteristics of the tumour and demonstrate that host malnutrition or tumour burden are not sufficient to drive wasting. We show that JAK/STAT and TNF-α/Egr signalling are elevated in cachectic muscle and promote tissue wasting. Furthermore, we introduce a dual driver system that allows independent genetic manipulation of tumour and host skeletal muscle. Overall, we present a novel Drosophila larval paradigm to study tumour/host tissue crosstalk in vivo, which may contribute to future research in cancer cachexia and impact the design of therapeutic approaches for this pathology.


Assuntos
Caquexia/patologia , Carcinogênese/patologia , Modelos Animais de Doenças , Larva/crescimento & desenvolvimento , Neoplasias/complicações , Animais , Caquexia/etiologia , Caquexia/metabolismo , Carcinogênese/genética , Carcinogênese/metabolismo , Drosophila , Perfilação da Expressão Gênica , Humanos , Janus Quinases/genética , Janus Quinases/metabolismo , Larva/genética , Larva/metabolismo , Fator de Transcrição STAT3/genética , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais
8.
Int J Mol Sci ; 22(16)2021 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-34445782

RESUMO

Spodoptera exigua is a worldwide pest afflicting edible vegetables and has developed varying levels of resistance to insecticides. Methoxyfenozide (MET), an ecdysteroid agonist, is effective against lepidopteran pests such as S. exigua. However, the mechanism of MET to S. exigua remains unclear. In this study, we analyzed the expression patterns of genes related to the ecdysone signaling pathway in transcriptome data treated with sublethal doses of MET and analyzed how expression levels of key genes affect the toxicity of MET on S. exigua. Our results demonstrated that 2639 genes were up-regulated and 2512 genes were down-regulated in S. exigua treated with LC30 of MET. Of these, 15 genes were involved in the ecdysone signaling pathway. qPCR results demonstrated that ecdysone receptor A (EcRA) expression levels significantly increased in S. exigua when treated with different doses of MET, and that the RNAi-mediated silencing of EcRA significantly increased mortality to 55.43% at 72 h when L3 S. exigua larvae were exposed to MET at the LC30 dose. Additionally, knocking down EcRA suppressed the most genes expressed in the ecdysone signaling pathway. The combination of MET and dsEcRA affected the expression of E74 and enhanced the expression of TREA. These results demonstrate that the adverse effects of sublethal MET disturb the ecdysone signaling pathway in S. exigua, and EcRA is closely related to MET toxic effect. This study increases our collective understanding of the mechanisms of MET in insect pests.


Assuntos
Ecdisona/genética , Hidrazinas/farmacologia , Hormônios Juvenis/farmacologia , Interferência de RNA/fisiologia , Transdução de Sinais/efeitos dos fármacos , Spodoptera/efeitos dos fármacos , Transcriptoma/genética , Animais , Perfilação da Expressão Gênica/métodos , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Larva/genética , Receptores de Esteroides/genética , Spodoptera/genética
9.
Ecotoxicol Environ Saf ; 223: 112574, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34358928

RESUMO

The widespread use of bisphenol S (BPS) as an alternative to bisphenol A has captured attention due to its potential toxicity to aquatic organisms. In the present study, the zebrafish was used as a model to evaluate the toxicity of BPS and determine the underlying mechanisms. The environmental concentration-dependent (0, 0.1, 1, 10, 100, and 1000 µg/L BPS) transcriptome approach was employed in combination with toxicity assays to address the problem. Based on a weighted correlation network analysis, we speculated that excess reactive oxygen species (ROS) may initiate cellular events in BPS-exposed zebrafish, leading to multiple toxic effects. Furthermore, we used pathway enrichment analysis to identify key pathways (MAPK signalling pathway and metabolic pathways) that link the molecular mechanisms with different toxic effects. In addition, we performed protein-protein network and shortest path analyses to identify six hub genes (erbb2, rrm2, rps27a, his2h3c, cdk1, and mcm5) and their interactions. Moreover, we suggest that BPS may interact with erbb2 by molecular docking. Thus, the BPS-erbb2 interaction may activate the MAPK signalling and metabolic pathways, resulting in ROS production and then caused multiple toxic effects in zebrafish. This study provides information for characterising the mechanisms of BPS exposure in aquatic environments.


Assuntos
Transcriptoma , Peixe-Zebra , Animais , Compostos Benzidrílicos/toxicidade , Larva/genética , Simulação de Acoplamento Molecular , Fenóis , Sulfonas , Peixe-Zebra/genética
10.
Pestic Biochem Physiol ; 178: 104934, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34446203

RESUMO

Chitin synthase (CHS) plays a critical role in chitin synthesis and excretion. In most insects, CHSs have been segregated into 1 and 2 classes. CHS1 is responsible for chitin production in the ectodermally-derived epidermal cells. CHS2 is dedicated to chitin biosynthesis in the midgut peritrophic matrix (PM). Henosepilachna vigintioctopunctata is a serious pest of Solanaceae and Cucurbitaceae plants. In this study, we identified HvCHS1 and HvCHS2. We found that HvCHS1 was abundantly transcribed in the larval tracheae and epidermis, whereas HvCHS2 was mainly expressed in the guts. Escherichia coli HT115 expressed double stranded RNAs targeting HvCHS1 and HvCHS2 (dsCHS1 and dsCHS2) were used to immerse potato foliage and the treated leaves were provided to the newly-molted fourth- and third-instar larvae. Ingestion of dsCHS1 by the fourth-instar larvae significantly diminished the target mRNA level and had slight influence on the expression of HvCHS2. In contrast, consumption of dsCHS2 significantly lowered the target mRNA level but triggered the transcription of HvCHS1. Knockdown of HvCHS1, rather than HvCHS2, arrested larval development and impaired larva-pupa-adult transition. A large proportion of HvCHS1 hypomorphs became stunting prepupae, deformed pupae or misshapen adults. Moreover, knockdown of HvCHS1 damaged gut integrity, decreased cuticle thickness, and delayed the formation of newly-generated cuticle layer during ecdysis. Furthermore, depletion of HvCHS1 inhibited the development of trachea system and thinned tracheal taenidia. Ingestion of dsCHS1 at the third-instar stage caused similar but severe negative effects. Our results demonstrated that HvCHS1 is responsible for chitin biosynthesis during ecdysis. Moreover, HvCHS1 is a potential amenable target gene and young larvae are more susceptible to dsRNA.


Assuntos
Quitina Sintase , Besouros , Animais , Quitina/metabolismo , Quitina Sintase/genética , Quitina Sintase/metabolismo , Besouros/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva/genética , Larva/metabolismo , Muda/genética , Pupa/metabolismo , Interferência de RNA
11.
Pestic Biochem Physiol ; 178: 104937, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34446204

RESUMO

For a devastating agricultural pest, functional genomics promotes the finding of novel technology to control Spodoptera frugiperda, such as the genetics-based strategies. In the present study, 11 yellow genes were identified in Spodoptera frugiperda. The transcriptome analysis showed the tissue-specific expression of part yellow genes, which suggested the importance of yellow genes in some biological processes in S. frugiperda, such as pigmentation. Among these yellow genes, the expression profiles of yellow-y gene showed that it was expressed in all life stages. In order to realize the further study of yellow-y, we employed CRISPR/Cas9 system to knock out this gene. Following knock out, diverse phenotypes were observed, such as color changes in both larvae and adults. Different from the wild-type larvae and adults, G0 mutants were yellowed since hatching. However, no color difference was observed with the pupal cuticle between the wild-type and mutant pupae before the 8th day. On the basis of the single-pair strategy of G0 generation, the yellow-y gene was proved to be a recessive gene. The G1 yellowish larvae with biallelic mutations displayed a relatively longer development period than wild-type, and often generated abnormal pupae and moths. The deletion of yellow-y also resulted in a decline in the fecundity. The results revealed that yellow-y gene was important for S. frugiperda pigmentation, as well as in its development and reproduction. Besides, the present study set up a standard procedure to knock out genes in S. frugiperda, which could be helpful for our understanding some key molecular processes, such as functional roles of detoxification genes as insecticide resistance mechanisms or modes of action of insecticides to facilitate the management of this insect pest.


Assuntos
Sistemas CRISPR-Cas , Mariposas , Animais , Sistemas CRISPR-Cas/genética , Resistência a Inseticidas , Larva/genética , Spodoptera/genética
12.
Pestic Biochem Physiol ; 178: 104943, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34446209

RESUMO

Structural cuticular proteins (CPs) are major components of the insect cuticle, and they play critical roles in insect development and insecticide resistance. Here, a total of 196 CP genes were successfully annotated in the Plutella xylostella genome. On the basis of motif analysis, these CPs were classified into 10 different families, including 122 CPR, 12 CPAP1, 8 CPAP3, 9 CPLCP, 2 Tweedle, 1 CPF, 1 CPFL, 1 CPCFC, 17 CPG and 2 18 aa proteins, and the remaining 21 unclassified CPs were classed as cuticular proteins hypothetical (CPH). A phylogenetic analysis of CPs from different insects revealed species-specific clades of RR-1 and RR-2 genes, suggesting that CP gene duplication might occur independently among insect taxa, while we also found that some other CPs (such as CPAP1 and CPAP3) had a closer relationship based on their conserved domain architecture. Using available RNAseq libraries, the expression profiles of the CPs were analyzed over the four developmental stages of the insect (i.e., egg, larva, pupa, and adult), revealing stage-specific expression patterns for the CPs. In a chlorpyrifos resistant strain, 18 CP genes were found to be more than two-fold upregulated compared to the susceptible control strain, and qRT-PCR analysis showed that these CP genes were overexpressed after exposure to chlorpyrifos, suggesting a potential role in the molecular mechanism of insecticide resistance in P. xylostella. This study provides the tools and molecular basis to study the role of CPs in the post-embryonal development and the mechanisms of insecticide resistance of P. xylostella.


Assuntos
Mariposas , Animais , Proteínas de Insetos/genética , Resistência a Inseticidas/genética , Larva/genética , Mariposas/genética , Filogenia
13.
Science ; 373(6554): 535-541, 2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34326235

RESUMO

Interkingdom competition occurs between hymenopteran parasitoids and insect viruses sharing the same insect hosts. It has been assumed that parasitoid larvae die with the death of the infected host or as result of competition for host resources. Here we describe a gene family, parasitoid killing factor (pkf), that encodes proteins toxic to parasitoids of the Microgastrinae group and determines parasitism success. Pkfs are found in several entomopathogenic DNA virus families and in some lepidopteran genomes. We provide evidence of equivalent and specific toxicity against endoparasites for PKFs found in entomopoxvirus, ascovirus, baculovirus, and Lepidoptera through a mechanism that elicits apoptosis in the cells of susceptible parasitoids. This highlights the evolutionary arms race between parasitoids, viruses, and their insect hosts.


Assuntos
Entomopoxvirinae/fisiologia , Proteínas de Insetos/toxicidade , Lepidópteros/parasitologia , Lepidópteros/virologia , Proteínas Virais/toxicidade , Vespas/fisiologia , Animais , Apoptose , Evolução Biológica , Transferência Genética Horizontal , Genoma de Inseto , Interações Hospedeiro-Parasita , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Vírus de Insetos/fisiologia , Larva/genética , Larva/parasitologia , Larva/virologia , Lepidópteros/genética , Lepidópteros/metabolismo , Nucleopoliedrovírus/fisiologia , Spodoptera/genética , Spodoptera/metabolismo , Spodoptera/parasitologia , Spodoptera/virologia , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/metabolismo , Vespas/crescimento & desenvolvimento
14.
Commun Biol ; 4(1): 860, 2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34253841

RESUMO

Schistosomes require both molluscan and mammalian hosts for development. The larval cercaria exits the snail host and swims to identify and invade the mammalian host. The cercaria has two macrostructures, the head and the tail. The head invades the host, where it matures into an adult worm. The tail is lost after host invasion. Translation in the cercaria differs in each macrostructure, with higher levels of translation in the cercarial tail and little to no translational activity in the cercarial head. We compared the transcriptome and proteome of the cercarial head and tail and observed stark differences between the two macrostructures. We identified unique and differentially expressed transcripts and proteins, including ribosomal components expressed in higher levels in tails than in heads, which may explain the differences in translation levels between heads and tails. We also characterized the weak correlation between transcription and translation in infectious cercarial heads and tails.


Assuntos
Proteínas de Helminto/metabolismo , Proteoma/metabolismo , Schistosoma mansoni/genética , Schistosoma mansoni/metabolismo , Transcriptoma/genética , Animais , Cercárias/genética , Cercárias/crescimento & desenvolvimento , Cercárias/metabolismo , Cromatografia Líquida/métodos , Perfilação da Expressão Gênica/métodos , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Proteômica/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Schistosoma mansoni/crescimento & desenvolvimento , Espectrometria de Massas em Tandem/métodos
15.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201772

RESUMO

Nucleolar stress occurs when ribosome production or function declines. Nucleolar stress in stem cells or progenitor cells often leads to disease states called ribosomopathies. Drosophila offers a robust system to explore how nucleolar stress causes cell cycle arrest, apoptosis, or autophagy depending on the cell type. We provide an overview of nucleolar stress in Drosophila by depleting nucleolar phosphoprotein of 140 kDa (Nopp140), a ribosome biogenesis factor (RBF) in nucleoli and Cajal bodies (CBs). The depletion of Nopp140 in eye imaginal disc cells generates eye deformities reminiscent of craniofacial deformities associated with the Treacher Collins syndrome (TCS), a human ribosomopathy. We show the activation of c-Jun N-terminal Kinase (JNK) in Drosophila larvae homozygous for a Nopp140 gene deletion. JNK is known to induce the expression of the pro-apoptotic Hid protein and autophagy factors Atg1, Atg18.1, and Atg8a; thus, JNK is a central regulator in Drosophila nucleolar stress. Ribosome abundance declines upon Nopp140 loss, but unusual cytoplasmic granules accumulate that resemble Processing (P) bodies based on marker proteins, Decapping Protein 1 (DCP1) and Maternal expression at 31B (Me31B). Wild type brain neuroblasts (NBs) express copious amounts of endogenous coilin, but coilin levels decline upon nucleolar stress in most NB types relative to the Mushroom body (MB) NBs. MB NBs exhibit resilience against nucleolar stress as they maintain normal coilin, Deadpan, and EdU labeling levels.


Assuntos
Nucléolo Celular/genética , Corpos Enovelados/patologia , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Ligação a RNA/genética , Estresse Fisiológico , Animais , Sistemas CRISPR-Cas , Corpos Enovelados/genética , Proteínas de Drosophila/antagonistas & inibidores , Drosophila melanogaster/fisiologia , Larva/genética , Larva/crescimento & desenvolvimento , Fosfoproteínas , Proteínas de Ligação a RNA/antagonistas & inibidores , Ribossomos/genética , Ribossomos/metabolismo
16.
Nat Commun ; 12(1): 3362, 2021 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-34099692

RESUMO

Diabetes can be caused by an insufficiency in ß-cell mass. Here, we performed a genetic screen in a zebrafish model of ß-cell loss to identify pathways promoting ß-cell regeneration. We found that both folate receptor 1 (folr1) overexpression and treatment with folinic acid, stimulated ß-cell differentiation in zebrafish. Treatment with folinic acid also stimulated ß-cell differentiation in cultures of neonatal pig islets, showing that the effect could be translated to a mammalian system. In both zebrafish and neonatal pig islets, the increased ß-cell differentiation originated from ductal cells. Mechanistically, comparative metabolomic analysis of zebrafish with/without ß-cell ablation and with/without folinic acid treatment indicated ß-cell regeneration could be attributed to changes in the pyrimidine, carnitine, and serine pathways. Overall, our results suggest evolutionarily conserved and previously unknown roles for folic acid and one-carbon metabolism in the generation of ß-cells.


Assuntos
Carbono/metabolismo , Diferenciação Celular/efeitos dos fármacos , Receptor 1 de Folato/metabolismo , Células Secretoras de Insulina/metabolismo , Leucovorina/farmacologia , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados , Animais Recém-Nascidos , Carnitina/metabolismo , Diferenciação Celular/genética , Células Cultivadas , Receptor 1 de Folato/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células Secretoras de Insulina/citologia , Larva/genética , Larva/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Camundongos , Pirimidinas/metabolismo , Suínos , Peixe-Zebra/genética
17.
J Agric Food Chem ; 69(26): 7429-7445, 2021 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-34169724

RESUMO

Tolerance to chemical insecticides can be driven by the necessity of herbivorous insects to defend against host plant-produced phytochemicals. However, how the phytochemicals are sensed and further transduced into a defense response associated with insecticide tolerance is poorly understood. Herein, we show that pre-exposure to flavone, a flavonoid phytochemical, effectively enhanced larval tolerance to multiple synthetic insecticides and elevated detoxification enzyme activities in Spodoptera litura. RNA-Seq analysis revealed that flavone induced a spectrum of genes spanning phase I and II detoxification enzyme families, as well as two transcription factors Cap "n" collar isoform C (CncC) and its partner small muscle aponeurosis fibromatosis (MafK). Knocking down of CncC by RNA interference suppressed flavone-induced detoxification gene expression and rendered the larvae more sensitive to the insecticides. Flavone exposure elicited a reactive oxygen species (ROS) burst, while scavenging of ROS inhibited CncC-mediated detoxification gene expression and suppressed flavone-induced detoxification enzyme activation. Metabolome analysis showed that the ingested flavone was mainly converted into three flavonoid metabolites, and only 3-hydroxyflavone was found to affect the ROS/CncC pathway-mediated metabolic detoxification. These results indicate that the ROS/CncC pathway is an important route driving detoxification gene expression responsible for insecticide tolerance after exposure to the phytochemical flavone.


Assuntos
Flavonas , Inseticidas , Animais , Humanos , Larva/genética , Compostos Fitoquímicos , Espécies Reativas de Oxigênio , Spodoptera/genética , Xenobióticos
18.
Methods Mol Biol ; 2326: 203-214, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34097270

RESUMO

Cyantraniliprole can effectively control lepidopteran pests and has been used all over the world. In general, the risk of cyantraniliprole seems low for fish, but the toxicity selectivity among different fish species was not clear. Here, we present the methods for the acute toxicity and chronic effects of cyantraniliprole by using juvenile tilapia (Oreochromis mossambicus). Based on this test, 96 h LC50 of cyantraniliprole to tilapia was 38.0 mg/L. After exposed for 28 days, specific growth rates of the blank control, solution control, and the treatments of 0.037, 0.37 and 3.7 mg/L of cyantraniliprole were 1.14, 0.95, 0.93, 0.82, and 0.70% per day, respectively. The results of micronucleus experiment and single cell gel electrophoresis showed that cyantraniliprole damaged DNA in liver cells of tilapia larvae. Quantitative PCR results showed that cyantraniliprole could induce the upregulation of Rpa 3 that is responsible for the DNA repair. The significant downregulation of Chk 2 gene was related to p53 pathway. It is therefore proposed that cyantraniliprole causes DNA damage in liver cells of tilapia and activates DNA damage and repair pathways.


Assuntos
Dano ao DNA/efeitos dos fármacos , Inseticidas/toxicidade , Pirazóis/toxicidade , Tilápia , Poluentes Químicos da Água/toxicidade , ortoaminobenzoatos/toxicidade , Animais , Larva/citologia , Larva/efeitos dos fármacos , Larva/genética , Larva/crescimento & desenvolvimento , Fígado/citologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Testes para Micronúcleos/métodos , Análise de Célula Única/métodos , Tilápia/crescimento & desenvolvimento , Tilápia/metabolismo , Testes de Toxicidade/métodos
19.
Mar Environ Res ; 169: 105379, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34119918

RESUMO

Cyclothone braueri (Stomiiformes, Gonostomatidae) is a widely distributed fish inhabiting the mesopelagic zone of marine tropical and temperate waters. Constituting one of the largest biomasses of the ocean, C. braueri is a key element in most of the ecological processes occurring in the twilight layer. We focused on the ecological processes linked to early life stages in relation to marine pelagic environmental drivers (temperature, salinity, food availability and geostrophic currents) considering different regions of the Central Mediterranean Sea. A multivariate morphometric analysis was carried out using six parameters with the aim of discerning different larval morphotypes, while a fragment of 367 bp representing the 12S ribosomal RNA gene was used to perform molecular analyses aimed at determining the intraspecific genetic variability. Analysis highlighted two geographically distinct morphotypes not genetically discernible and related to the different nutritional conditions due to spatial heterogeneities in terms of temperature and food availability. The body depth (BD) emerged as an appropriate morphometric parameter to detect the larval condition in this species. Molecular analysis highlighted a moderate genetic divergence in the fish population, showing the recurrence of two phylogroups not geographically separated.


Assuntos
Cilióforos , Peixes , Animais , Estruturas Genéticas , Larva/genética , Mar Mediterrâneo
20.
Commun Biol ; 4(1): 649, 2021 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-34059788

RESUMO

The cryptic parasite Sparganum proliferum proliferates in humans and invades tissues and organs. Only scattered cases have been reported, but S. proliferum infection is always fatal. However, S. proliferum's phylogeny and life cycle remain enigmatic. To investigate the phylogenetic relationships between S. proliferum and other cestode species, and to examine the mechanisms underlying pathogenicity, we sequenced the entire genomes of S. proliferum and a closely related non-life-threatening tapeworm Spirometra erinaceieuropaei. Additionally, we performed larvae transcriptome analyses of S. proliferum plerocercoid to identify genes involved in asexual reproduction in the host. The genome sequences confirmed that the S. proliferum has experienced a clearly distinct evolutionary history from S. erinaceieuropaei. Moreover, we found that nonordinal extracellular matrix coordination allows asexual reproduction in the host, and loss of sexual maturity in S. proliferum are responsible for its fatal pathogenicity to humans. Our high-quality reference genome sequences should be valuable for future studies of pseudophyllidean tapeworm biology and parasitism.


Assuntos
Plerocercoide/genética , Animais , Sequência de Bases/genética , Proliferação de Células/genética , Cestoides/classificação , Cestoides/genética , Infecções por Cestoides/genética , Infecções por Cestoides/parasitologia , Genoma/genética , Humanos , Larva/classificação , Larva/genética , Estágios do Ciclo de Vida/genética , Filogenia , Plerocercoide/classificação , Spirometra/classificação , Spirometra/genética
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