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1.
Environ Pollut ; 294: 118646, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-34896224

RESUMO

With the rapidly increasing popularity of 5G mobile technology, the effect of radiofrequency radiation on human health has caused public concern. This study explores the effects of a simulated 3.5 GHz radiofrequency electromagnetic radiation (RF-EMF) environment on the development and microbiome of flies under intensities of 0.1 W/m2, 1 W/m2 and 10 W/m2. We found that the pupation percentages in the first 3 days and eclosion rate in the first 2 days were increased under exposure to RF-EMF, and the mean development time was shortened. In a study on third-instar larvae, the expression levels of the heat shock protein genes hsp22, hsp26 and hsp70 and humoral immune system genes AttC, TotC and TotA were all significantly increased. In the oxidative stress system, DuoX gene expression was decreased, sod2 and cat gene expression levels were increased, and SOD and CAT enzyme activity also showed a significant increase. According to the 16S rDNA results, the diversity and species abundance of the microbial community decreased significantly, and according to the functional prediction analysis, the genera Acetobacter and Lactobacillus were significantly increased. In conclusion, 3.5 GHz RF-EMF may enhance thermal stress, oxidative stress and humoral immunity, cause changes in the microbial community, and regulate the insulin/TOR and ecdysteroid signalling pathways to promote fly development.


Assuntos
Drosophila melanogaster , Campos Eletromagnéticos , Microbiota/efeitos da radiação , Ondas de Rádio , Animais , Telefone Celular , Drosophila melanogaster/embriologia , Drosophila melanogaster/microbiologia , Drosophila melanogaster/efeitos da radiação , Expressão Gênica , Proteínas de Choque Térmico , Larva/efeitos da radiação
2.
Elife ; 102021 10 22.
Artigo em Inglês | MEDLINE | ID: mdl-34677126

RESUMO

Wolbachia are the most widespread bacterial endosymbionts in animals. Within arthropods, these maternally transmitted bacteria can selfishly hijack host reproductive processes to increase the relative fitness of their transmitting females. One such form of reproductive parasitism called male killing, or the selective killing of infected males, is recapitulated to degrees by transgenic expression of the prophage WO-mediated killing (wmk) gene. Here, we characterize the genotype-phenotype landscape of wmk-induced male killing in D. melanogaster using transgenic expression. While phylogenetically distant wmk homologs induce no sex-ratio bias, closely-related homologs exhibit complex phenotypes spanning no death, male death, or death of all hosts. We demonstrate that alternative start codons, synonymous codons, and notably a single synonymous nucleotide in wmk can ablate killing. These findings reveal previously unrecognized features of transgenic wmk-induced killing and establish new hypotheses for the impacts of post-transcriptional processes in male killing variation. We conclude that synonymous sequence changes are not necessarily silent in nested endosymbiotic interactions with life-or-death consequences.


Assuntos
Proteínas de Bactérias/genética , Drosophila melanogaster/microbiologia , Prófagos/genética , Simbiose , Wolbachia/fisiologia , Animais , Proteínas de Bactérias/metabolismo , Masculino , Microrganismos Geneticamente Modificados/genética , Microrganismos Geneticamente Modificados/fisiologia , Wolbachia/genética
3.
Proc Natl Acad Sci U S A ; 118(39)2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34548405

RESUMO

Animals interact with microbes that affect their performance and fitness, including endosymbionts that reside inside their cells. Maternally transmitted Wolbachia bacteria are the most common known endosymbionts, in large part because of their manipulation of host reproduction. For example, many Wolbachia cause cytoplasmic incompatibility (CI) that reduces host embryonic viability when Wolbachia-modified sperm fertilize uninfected eggs. Operons termed cifs control CI, and a single factor (cifA) rescues it, providing Wolbachia-infected females a fitness advantage. Despite CI's prevalence in nature, theory indicates that natural selection does not act to maintain CI, which varies widely in strength. Here, we investigate the genetic and functional basis of CI-strength variation observed among sister Wolbachia that infect Drosophila melanogaster subgroup hosts. We cloned, Sanger sequenced, and expressed cif repertoires from weak CI-causing wYak in Drosophila yakuba, revealing mutations suspected to weaken CI relative to model wMel in D. melanogaster A single valine-to-leucine mutation within the deubiquitylating (DUB) domain of the wYak cifB homolog (cidB) ablates a CI-like phenotype in yeast. The same mutation reduces both DUB efficiency in vitro and transgenic CI strength in the fly, each by about twofold. Our results map hypomorphic transgenic CI to reduced DUB activity and indicate that deubiquitylation is central to CI induction in cid systems. We also characterize effects of other genetic variation distinguishing wMel-like cifs Importantly, CI strength determines Wolbachia prevalence in natural systems and directly influences the efficacy of Wolbachia biocontrol strategies in transinfected mosquito systems. These approaches rely on strong CI to reduce human disease.


Assuntos
Citoplasma/patologia , Drosophila melanogaster/microbiologia , Embrião não Mamífero/microbiologia , Mutação , Simbiose , Ubiquitinação , Wolbachia/fisiologia , Animais , Citoplasma/microbiologia , Enzimas Desubiquitinantes/metabolismo , Drosophila melanogaster/genética , Embrião não Mamífero/metabolismo , Feminino , Masculino
4.
Int J Mol Sci ; 22(18)2021 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-34576280

RESUMO

Avoiding excessive or insufficient immune responses and maintaining homeostasis are critical for animal survival. Although many positive or negative modulators involved in immune responses have been identified, little has been reported to date concerning whether the long non-coding RNA (lncRNA) can regulate Drosophila immunity response. In this study, we firstly discover that the overexpression of lncRNA-CR11538 can inhibit the expressions of antimicrobial peptides Drosomycin (Drs) and Metchnikowin (Mtk) in vivo, thereby suppressing the Toll signaling pathway. Secondly, our results demonstrate that lncRNA-CR11538 can interact with transcription factors Dif/Dorsal in the nucleus based on both subcellular localization and RIP analyses. Thirdly, our findings reveal that lncRNA-CR11538 can decoy Dif/Dorsal away from the promoters of Drs and Mtk to repress their transcriptions by ChIP-qPCR and dual luciferase report experiments. Fourthly, the dynamic expression changes of Drs, Dif, Dorsal and lncRNA-CR11538 in wild-type flies (w1118) at different time points after M. luteus stimulation disclose that lncRNA-CR11538 can help Drosophila restore immune homeostasis in the later period of immune response. Overall, our study reveals a novel mechanism by which lncRNA-CR11538 serves as a Dif/Dorsal decoy to downregulate antimicrobial peptide expressions for restoring Drosophila Toll immunity homeostasis, and provides a new insight into further studying the complex regulatory mechanism of animal innate immunity.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/imunologia , Drosophila melanogaster/metabolismo , Micrococcus luteus , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Proteínas Citotóxicas Formadoras de Poros/farmacologia , RNA Longo não Codificante , Fatores de Transcrição/metabolismo , Animais , Sítios de Ligação , Proteínas de Ligação a DNA/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/microbiologia , Homeostase , Imunidade Inata , Masculino , NF-kappa B/metabolismo , Proteínas Nucleares/genética , Fosfoproteínas/genética , Regiões Promotoras Genéticas , RNA-Seq , Sepse , Transdução de Sinais , Frações Subcelulares , Receptores Toll-Like/metabolismo , Fatores de Transcrição/genética
5.
PLoS Pathog ; 17(8): e1009846, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34432851

RESUMO

The fruit fly Drosophila melanogaster combats microbial infection by producing a battery of effector peptides that are secreted into the haemolymph. Technical difficulties prevented the investigation of these short effector genes until the recent advent of the CRISPR/CAS era. As a consequence, many putative immune effectors remain to be formally described, and exactly how each of these effectors contribute to survival is not well characterized. Here we describe a novel Drosophila antifungal peptide gene that we name Baramicin A. We show that BaraA encodes a precursor protein cleaved into multiple peptides via furin cleavage sites. BaraA is strongly immune-induced in the fat body downstream of the Toll pathway, but also exhibits expression in other tissues. Importantly, we show that flies lacking BaraA are viable but susceptible to the entomopathogenic fungus Beauveria bassiana. Consistent with BaraA being directly antimicrobial, overexpression of BaraA promotes resistance to fungi and the IM10-like peptides produced by BaraA synergistically inhibit growth of fungi in vitro when combined with a membrane-disrupting antifungal. Surprisingly, BaraA mutant males but not females display an erect wing phenotype upon infection. Here, we characterize a new antifungal immune effector downstream of Toll signalling, and show it is a key contributor to the Drosophila antimicrobial response.


Assuntos
Antifúngicos/farmacologia , Beauveria/efeitos dos fármacos , Proteínas de Drosophila/farmacologia , Drosophila melanogaster/efeitos dos fármacos , Micoses/tratamento farmacológico , Peptídeos/farmacologia , Animais , Beauveria/crescimento & desenvolvimento , Beauveria/imunologia , Drosophila melanogaster/genética , Drosophila melanogaster/imunologia , Drosophila melanogaster/microbiologia , Feminino , Masculino , Micoses/imunologia , Micoses/microbiologia
6.
PLoS Pathog ; 17(8): e1009326, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34339477

RESUMO

Metabolic pathways are now considered as intrinsic virulence attributes of pathogenic bacteria and thus represent potential targets for antibacterial strategies. Here we focused on the role of the pentose phosphate pathway (PPP) and its connections with other metabolic pathways in the pathophysiology of Francisella novicida. The involvement of the PPP in the intracellular life cycle of Francisella was first demonstrated by studying PPP inactivating mutants. Indeed, we observed that inactivation of the tktA, rpiA or rpe genes severely impaired intramacrophage multiplication during the first 24 hours. However, time-lapse video microscopy demonstrated that rpiA and rpe mutants were able to resume late intracellular multiplication. To better understand the links between PPP and other metabolic networks in the bacterium, we also performed an extensive proteo-metabolomic analysis of these mutants. We show that the PPP constitutes a major bacterial metabolic hub with multiple connections to glycolysis, the tricarboxylic acid cycle and other pathways, such as fatty acid degradation and sulfur metabolism. Altogether our study highlights how PPP plays a key role in the pathogenesis and growth of Francisella in its intracellular niche.


Assuntos
Proteínas de Bactérias/metabolismo , Drosophila melanogaster/metabolismo , Francisella/patogenicidade , Infecções por Bactérias Gram-Negativas/microbiologia , Metaboloma , Via de Pentose Fosfato , Proteoma , Animais , Proteínas de Bactérias/genética , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/microbiologia , Francisella/metabolismo , Regulação Bacteriana da Expressão Gênica , Glicólise , Macrófagos/metabolismo , Macrófagos/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mutação
7.
PLoS Pathog ; 17(8): e1009859, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34383852

RESUMO

Wolbachia is a group of intracellular symbiotic bacteria that widely infect arthropods and nematodes. Wolbachia infection can regulate host reproduction with the most common phenotype in insects being cytoplasmic incompatibility (CI), which results in embryonic lethality when uninfected eggs fertilized with sperms from infected males. This suggests that CI-induced defects are mainly in paternal side. However, whether Wolbachia-induced metabolic changes play a role in the mechanism of paternal-linked defects in embryonic development is not known. In the current study, we first use untargeted metabolomics method with LC-MS to explore how Wolbachia infection influences the metabolite profiling of the insect hosts. The untargeted metabolomics revealed 414 potential differential metabolites between Wolbachia-infected and uninfected 1-day-old (1d) male flies. Most of the differential metabolites were significantly up-regulated due to Wolbachia infection. Thirty-four metabolic pathways such as carbohydrate, lipid and amino acid, and vitamin and cofactor metabolism were affected by Wolbachia infection. Then, we applied targeted metabolomics analysis with GC-MS and showed that Wolbachia infection resulted in an increased energy expenditure of the host by regulating glycometabolism and fatty acid catabolism, which was compensated by increased food uptake. Furthermore, overexpressing two acyl-CoA catabolism related genes, Dbi (coding for diazepam-binding inhibitor) or Mcad (coding for medium-chain acyl-CoA dehydrogenase), ubiquitously or specially in testes caused significantly decreased paternal-effect egg hatch rate. Oxidative stress and abnormal mitochondria induced by Wolbachia infection disrupted the formation of sperm nebenkern. These findings provide new insights into mechanisms of Wolbachia-induced paternal defects from metabolic phenotypes.


Assuntos
Infecções Bacterianas/complicações , Drosophila melanogaster/metabolismo , Infertilidade Masculina/patologia , Metaboloma , Fenótipo , Reprodução , Wolbachia/fisiologia , Animais , Infecções Bacterianas/metabolismo , Infecções Bacterianas/microbiologia , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/microbiologia , Feminino , Infertilidade Masculina/etiologia , Infertilidade Masculina/metabolismo , Masculino
8.
Nature ; 596(7870): 97-102, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34290404

RESUMO

Infection-induced aversion against enteropathogens is a conserved sickness behaviour that can promote host survival1,2. The aetiology of this behaviour remains poorly understood, but studies in Drosophila have linked olfactory and gustatory perception to avoidance behaviours against toxic microorganisms3-5. Whether and how enteric infections directly influence sensory perception to induce or modulate such behaviours remains unknown. Here we show that enteropathogen infection in Drosophila can modulate olfaction through metabolic reprogramming of ensheathing glia of the antennal lobe. Infection-induced unpaired cytokine expression in the intestine activates JAK-STAT signalling in ensheathing glia, inducing the expression of glial monocarboxylate transporters and the apolipoprotein glial lazarillo (GLaz), and affecting metabolic coupling of glia and neurons at the antennal lobe. This modulates olfactory discrimination, promotes the avoidance of bacteria-laced food and increases fly survival. Although transient in young flies, gut-induced metabolic reprogramming of ensheathing glia becomes constitutive in old flies owing to age-related intestinal inflammation, which contributes to an age-related decline in olfactory discrimination. Our findings identify adaptive glial metabolic reprogramming by gut-derived cytokines as a mechanism that causes lasting changes in a sensory system in ageing flies.


Assuntos
Envelhecimento/metabolismo , Citocinas/metabolismo , Drosophila melanogaster/metabolismo , Intestinos , Neuroglia/metabolismo , Olfato/fisiologia , Animais , Aprendizagem da Esquiva , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/microbiologia , Feminino , Inflamação/metabolismo , Inflamação/microbiologia , Intestinos/microbiologia , Janus Quinases/metabolismo , Ácido Láctico/metabolismo , Metabolismo dos Lipídeos , Neurônios/metabolismo , Pectobacterium carotovorum , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais , Taxa de Sobrevida , Fatores de Transcrição/metabolismo
9.
PLoS Genet ; 17(6): e1009612, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34143770

RESUMO

Wolbachia is one of the most prevalent bacterial endosymbionts, infecting approximately 40% of terrestrial arthropod species. Wolbachia is often a reproductive parasite but can also provide fitness benefits to its host, as, for example, protection against viral pathogens. This protective effect is currently being applied to fight arboviruses transmission by releasing Wolbachia-transinfected mosquitoes. Titre regulation is a crucial aspect of Wolbachia biology. Higher titres can lead to stronger phenotypes and fidelity of transmission but can have a higher cost to the host. Since Wolbachia is maternally transmitted, its fitness depends on host fitness, and, therefore, its cost to the host may be under selection. Understanding how Wolbachia titres are regulated and other aspects of Wolbachia biology has been hampered by the lack of genetic tools. Here we developed a forward genetic screen to identify new Wolbachia over-proliferative mutant variants. We characterized in detail two new mutants, wMelPop2 and wMelOctoless, and show that the amplification or loss of the Octomom genomic region lead to over-proliferation. These results confirm previous data and expand on the complex role of this genomic region in the control of Wolbachia proliferation. Both new mutants shorten the host lifespan and increase antiviral protection. Moreover, we show that Wolbachia proliferation rate in Drosophila melanogaster depends on the interaction between Octomom copy number, the host developmental stage, and temperature. Our analysis also suggests that the life shortening and antiviral protection phenotypes of Wolbachia are dependent on different, but related, properties of the endosymbiont; the rate of proliferation and the titres near the time of infection, respectively. We also demonstrate the feasibility of a novel and unbiased experimental approach to study Wolbachia biology, which could be further adapted to characterize other genetically intractable bacterial endosymbionts.


Assuntos
Drosophila melanogaster/microbiologia , Genoma Bacteriano , Longevidade/imunologia , Simbiose/genética , Wolbachia/genética , Animais , Carga Bacteriana , Dicistroviridae/crescimento & desenvolvimento , Dicistroviridae/patogenicidade , Drosophila melanogaster/imunologia , Drosophila melanogaster/virologia , Feminino , Edição de Genes/métodos , Ilhas Genômicas , Masculino , Fenótipo , Wolbachia/crescimento & desenvolvimento , Wolbachia/metabolismo
10.
mBio ; 12(3): e0027621, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34126772

RESUMO

Antibiotic-resistant Staphylococcus aureus strains constitute a major public health concern worldwide and are responsible for both health care- and community-associated infections. Here, we establish a robust and easy-to-implement model of oral S. aureus infection using Drosophila melanogaster larvae that allowed us to follow the fate of S. aureus at the whole-organism level as well as the host immune responses. Our study demonstrates that S. aureus infection triggers H2O2 production by the host via the Duox enzyme, thereby promoting antimicrobial peptide production through activation of the Toll pathway. Staphylococcal catalase mediates H2O2 neutralization, which not only promotes S. aureus survival but also minimizes the host antimicrobial response, hence reducing bacterial clearance in vivo. We show that while catalase expression is regulated in vitro by the accessory gene regulatory system (Agr) and the general stress response regulator sigma B (SigB), it no longer depends on these two master regulators in vivo. Finally, we confirm the versatility of this model by demonstrating the colonization and host stimulation capabilities of S. aureus strains belonging to different sequence types (CC8 and CC5) as well as of two other bacterial pathogens, Salmonella enterica serovar Typhimurium and Shigella flexneri. Thus, the Drosophila larva can be a general model to follow in vivo the innate host immune responses triggered during infection by human pathogens. IMPORTANCE The pathogenicity of methicillin-resistant S. aureus (MRSA) strains relies on their ability to produce a wide variety of tightly regulated virulence factors. Current in vivo models to analyze host-pathogen interactions are limited and difficult to manipulate. Here, we have established a robust and reliable model of oral S. aureus infection using Drosophila melanogaster larvae. We show that S. aureus stimulates host immunity through the production of reactive oxygen species (ROS) and antimicrobial peptide (AMP) and that ROS potentialize AMP gene expression. S. aureus catalase plays a key role in this complex environment and acts in vivo independently from SigB and Agr control. We propose that fly larvae can provide a general model for studying the colonization capabilities of human pathogens.


Assuntos
Interações Hospedeiro-Patógeno/imunologia , Imunidade Inata , Staphylococcus aureus Resistente à Meticilina/imunologia , Staphylococcus aureus Resistente à Meticilina/patogenicidade , Espécies Reativas de Oxigênio/imunologia , Animais , Modelos Animais de Doenças , Drosophila melanogaster/imunologia , Drosophila melanogaster/microbiologia , Regulação Bacteriana da Expressão Gênica , Larva/imunologia , Larva/microbiologia , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Citotóxicas Formadoras de Poros/imunologia , Espécies Reativas de Oxigênio/metabolismo , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/microbiologia , Virulência
11.
Immunity ; 54(8): 1683-1697.e3, 2021 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-34107298

RESUMO

Microbe-derived acetate activates the Drosophila immunodeficiency (IMD) pathway in a subset of enteroendocrine cells (EECs) of the anterior midgut. In these cells, the IMD pathway co-regulates expression of antimicrobial and enteroendocrine peptides including tachykinin, a repressor of intestinal lipid synthesis. To determine whether acetate acts on a cell surface pattern recognition receptor or an intracellular target, we asked whether acetate import was essential for IMD signaling. Mutagenesis and RNA interference revealed that the putative monocarboxylic acid transporter Tarag was essential for enhancement of IMD signaling by dietary acetate. Interference with histone deacetylation in EECs augmented transcription of genes regulated by the steroid hormone ecdysone including IMD targets. Reduced expression of the histone acetyltransferase Tip60 decreased IMD signaling and blocked rescue by dietary acetate and other sources of intracellular acetyl-CoA. Thus, microbe-derived acetate induces chromatin remodeling within enteroendocrine cells, co-regulating host metabolism and intestinal innate immunity via a Tip60-steroid hormone axis that is conserved in mammals.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/imunologia , Células Enteroendócrinas/metabolismo , Microbioma Gastrointestinal/imunologia , Histona Acetiltransferases/metabolismo , Intestinos/imunologia , Acetatos/imunologia , Acetilcoenzima A/metabolismo , Animais , Montagem e Desmontagem da Cromatina/fisiologia , Drosophila melanogaster/microbiologia , Ecdisona/metabolismo , Imunidade Inata/imunologia , Intestinos/microbiologia , Transportadores de Ácidos Monocarboxílicos/genética , Transportadores de Ácidos Monocarboxílicos/metabolismo , Interferência de RNA , Transdução de Sinais/imunologia , Taquicininas/metabolismo
12.
Sci Rep ; 11(1): 11336, 2021 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-34059765

RESUMO

Wolbachia are maternally inherited endosymbionts that infect nearly half of all arthropod species. Wolbachia manipulate their hosts to maximize their transmission, but they can also provide benefits such as nutrients and resistance against viruses to their hosts. The Wolbachia strain wMel was recently found to increase locomotor activities and possibly trigger cytoplasmic incompatibility in the transinfected fly Drosophila nigrosparsa. Here, we investigated, in females of both D. melanogaster and D. nigrosparsa, the gene expression between animals uninfected and infected with wMel, using RNA sequencing to see if the two Drosophila species respond to the infection in the same or different ways. A total of 2164 orthologous genes were used. The two fly species responded to the infection in different ways. Significant changes shared by the fly species belong to the expression of genes involved in processes such as oxidation-reduction process, iron-ion binding, and voltage-gated potassium-channel activity. We discuss our findings also in the light of how Wolbachia survive within both the native and the novel host.


Assuntos
Drosophila melanogaster/metabolismo , Drosophila melanogaster/microbiologia , Expressão Gênica , Wolbachia/fisiologia , Animais , Feminino , Perfilação da Expressão Gênica , Especificidade da Espécie , Simbiose
13.
Nat Commun ; 12(1): 2698, 2021 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-33976215

RESUMO

Gut microbiome profoundly affects many aspects of host physiology and behaviors. Here we report that gut microbiome modulates aggressive behaviors in Drosophila. We found that germ-free males showed substantial decrease in inter-male aggression, which could be rescued by microbial re-colonization. These germ-free males are not as competitive as wild-type males for mating with females, although they displayed regular levels of locomotor and courtship behaviors. We further found that Drosophila microbiome interacted with diet during a critical developmental period for the proper expression of octopamine and manifestation of aggression in adult males. These findings provide insights into how gut microbiome modulates specific host behaviors through interaction with diet during development.


Assuntos
Agressão/fisiologia , Drosophila melanogaster/fisiologia , Microbioma Gastrointestinal/fisiologia , Octopamina/metabolismo , Comportamento Sexual Animal/fisiologia , Animais , Bactérias/classificação , Bactérias/genética , Encéfalo/citologia , Encéfalo/metabolismo , Encéfalo/fisiologia , Drosophila melanogaster/metabolismo , Drosophila melanogaster/microbiologia , Feminino , Microbioma Gastrointestinal/genética , Masculino , Atividade Motora/fisiologia , Neurônios/metabolismo , Neurônios/fisiologia , RNA Ribossômico 16S/genética , Transdução de Sinais/fisiologia , Organismos Livres de Patógenos Específicos
14.
BMC Microbiol ; 21(1): 108, 2021 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-33836662

RESUMO

BACKGROUND: Experimental evolution has a long history of uncovering fundamental insights into evolutionary processes, but has largely neglected one underappreciated component--the microbiome. As eukaryotic hosts evolve, the microbiome may also respond to selection. However, the microbial contribution to host evolution remains poorly understood. Here, we re-analyzed genomic data to characterize the metagenomes from ten Evolve and Resequence (E&R) experiments in Drosophila melanogaster to determine how the microbiome changed in response to host selection. RESULTS: Bacterial diversity was significantly different in 5/10 studies, primarily in traits associated with metabolism or immunity. Duration of selection did not significantly influence bacterial diversity, highlighting the importance of associations with specific host traits. CONCLUSIONS: Our genomic re-analysis suggests the microbiome often responds to host selection; thus, the microbiome may contribute to the response of Drosophila in E&R experiments. We outline important considerations for incorporating the microbiome into E&R experiments. The E&R approach may provide critical insights into host-microbiome interactions and fundamental insight into the genomic basis of adaptation.


Assuntos
Drosophila melanogaster/microbiologia , Evolução Molecular , Interações entre Hospedeiro e Microrganismos/fisiologia , Adaptação Fisiológica , Animais , Bactérias/genética , Biodiversidade , Microbiota/genética , Seleção Genética
15.
Int J Mol Sci ; 22(5)2021 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-33800390

RESUMO

The fruit fly, Drosophila melanogaster, has been used to understand fundamental principles of genetics and biology for over a century. Drosophila is now also considered an essential tool to study mechanisms underlying numerous human genetic diseases. In this review, we will discuss how flies can be used to deepen our knowledge of infectious disease mechanisms in vivo. Flies make effective and applicable models for studying host-pathogen interactions thanks to their highly conserved innate immune systems and cellular processes commonly hijacked by pathogens. Drosophila researchers also possess the most powerful, rapid, and versatile tools for genetic manipulation in multicellular organisms. This allows for robust experiments in which specific pathogenic proteins can be expressed either one at a time or in conjunction with each other to dissect the molecular functions of each virulent factor in a cell-type-specific manner. Well documented phenotypes allow large genetic and pharmacological screens to be performed with relative ease using huge collections of mutant and transgenic strains that are publicly available. These factors combine to make Drosophila a powerful tool for dissecting out host-pathogen interactions as well as a tool to better understand how we can treat infectious diseases that pose risks to public health, including COVID-19, caused by SARS-CoV-2.


Assuntos
Doenças Transmissíveis/imunologia , Doenças Transmissíveis/metabolismo , Drosophila melanogaster/imunologia , Drosophila melanogaster/metabolismo , Animais , Doenças Transmissíveis/microbiologia , Doenças Transmissíveis/virologia , Drosophila melanogaster/microbiologia , Drosophila melanogaster/virologia , Interações Hospedeiro-Patógeno , Imunidade Inata , Transdução de Sinais , Fatores de Virulência/metabolismo
16.
Elife ; 102021 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-33879316

RESUMO

Reduced activity of the insulin/IGF signalling network increases health during ageing in multiple species. Diverse and tissue-specific mechanisms drive the health improvement. Here, we performed tissue-specific transcriptional and proteomic profiling of long-lived Drosophila dilp2-3,5 mutants, and identified tissue-specific regulation of >3600 transcripts and >3700 proteins. Most expression changes were regulated post-transcriptionally in the fat body, and only in mutants infected with the endosymbiotic bacteria, Wolbachia pipientis, which increases their lifespan. Bioinformatic analysis identified reduced co-translational ER targeting of secreted and membrane-associated proteins and increased DNA damage/repair response proteins. Accordingly, age-related DNA damage and genome instability were lower in fat body of the mutant, and overexpression of a minichromosome maintenance protein subunit extended lifespan. Proteins involved in carbohydrate metabolism showed altered expression in the mutant intestine, and gut-specific overexpression of a lysosomal mannosidase increased autophagy, gut homeostasis, and lifespan. These processes are candidates for combatting ageing-related decline in other organisms.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Insulina/fisiologia , Proteoma/metabolismo , Transdução de Sinais , Transcriptoma , Wolbachia/fisiologia , Animais , Drosophila melanogaster/metabolismo , Drosophila melanogaster/microbiologia , Corpo Adiposo/metabolismo , Perfilação da Expressão Gênica
17.
PLoS One ; 16(4): e0250524, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33914801

RESUMO

Insects are frequently infected with heritable bacterial endosymbionts. Endosymbionts have a dramatic impact on their host physiology and evolution. Their tissue distribution is variable with some species being housed intracellularly, some extracellularly and some having a mixed lifestyle. The impact of extracellular endosymbionts on the biofluids they colonize (e.g. insect hemolymph) is however difficult to appreciate because biofluid composition can depend on the contribution of numerous tissues. Here we investigate Drosophila hemolymph proteome changes in response to the infection with the endosymbiont Spiroplasma poulsonii. S. poulsonii inhabits the fly hemolymph and gets vertically transmitted over generations by hijacking the oogenesis in females. Using dual proteomics on infected hemolymph, we uncovered a weak, chronic activation of the Toll immune pathway by S. poulsonii that was previously undetected by transcriptomics-based approaches. Using Drosophila genetics, we also identified candidate proteins putatively involved in controlling S. poulsonii growth. Last, we also provide a deep proteome of S. poulsonii, which, in combination with previously published transcriptomics data, improves our understanding of the post-transcriptional regulations operating in this bacterium.


Assuntos
Drosophila melanogaster/genética , Proteoma/genética , Proteômica , Spiroplasma/genética , Animais , Proteínas de Bactérias/genética , Drosophila melanogaster/imunologia , Drosophila melanogaster/microbiologia , Feminino , Hemolinfa/microbiologia , Oogênese/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Spiroplasma/patogenicidade , Simbiose/genética , Simbiose/imunologia
18.
Lett Appl Microbiol ; 73(1): 31-38, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33735468

RESUMO

Candida species are opportunistic human fungal pathogens that cause acute and chronic infections against which only few antifungal agents are available. Here we have elucidated the antifungal effect of Syzygium samarangense leaf extracts (SSLE). Antifungal activity of SSLE was studied against Candida albicans, C. krusei, C. parapsilosis, C. glabrata, C. auris and C. tropicalis. Following experiments were performed: minimum fungicidal concentration (MFC) determination, agar well disc diffusion assays, fungal morphology analysis using scanning electron microscope (SEM), ex vivo fungal survival assays on porcine tongue and skin and in vivo fungal survival assays using Drosophila melanogaster fly model. Results demonstrated MFC of SSLE ranges between 100 and 125 mg ml-1 . SEM images showed cell wall degradation of C. albicans when treated with SSLE. Around 75% decrease in C. albicans viability was observed when infected porcine tongue and skin were treated using SSLE. The C. albicans infected D. melanogaster when fed with SSLE showed significant decrease (around 80%) of fungal count than the infected control. Furthermore, agar plate disc diffusion assays demonstrated that the antifungal activity of SSLE could be due to chalcone, which is one of the active constituents in SSLE. Our study demonstrated that SSLE could be used for the topical treatment of Candida infections.


Assuntos
Candida/efeitos dos fármacos , Extratos Vegetais/farmacologia , Syzygium/química , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candidíase/tratamento farmacológico , Drosophila melanogaster/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Extratos Vegetais/uso terapêutico , Suínos
19.
Arch Insect Biochem Physiol ; 106(4): e21776, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33644932

RESUMO

Maternally inherited endosymbiotic bacterium Wolbachia infects Drosophila melanogaster populations worldwide. Its genetic diversity includes several closely related genotypes, which can be attributed to two main genotype groups: wMel and wMelCS. Here, we studied eight D. melanogaster lines carrying the nuclear background of wild type interbred Bi90 line and cytoplasmic backgrounds with or without Wolbachia of different origin, each of which belongs to wMelCS genotype group. We analyzed the effect these seven Wolbachia strains had on the heat stress resistance and dopamine metabolism in D. melanogaster females. Survival under heat stress (38°C, 3 h 30 min) was increased only in the line infected with bacteria of the wMelPlus strain. At the same time, the activity of alkaline phosphatase (an enzyme regulating the pool of dopamine precursor tyrosine) was increased under normal conditions in females infected with all strains under study and retained the response to heat stress typical for the uninfected line. Thus, we found the unique Wolbachia strain that provides an increase of the host stress resistance, and demonstrated that the mechanism of this resistance is not dopamine-mediated.


Assuntos
Drosophila melanogaster/microbiologia , Termotolerância , Wolbachia/genética , Fosfatase Alcalina/metabolismo , Animais , Dopamina/metabolismo , Drosophila melanogaster/metabolismo , Genótipo , Resposta ao Choque Térmico , Simbiose/fisiologia , Termotolerância/genética , Termotolerância/fisiologia
20.
PLoS One ; 16(2): e0247376, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33606846

RESUMO

All metazoans are colonized by a complex and diverse set of microorganisms. The microbes colonize all parts of the body and are especially abundant in the gastrointestinal tract, where they constitute the gut microbiome. The fruit fly Drosophila melanogaster turned out to be an exquisite model organism to functionally test the importance of an intact gut microbiome. Still, however, fundamental questions remain unanswered. For example, it is unknown whether a fine-tuned regionalization of the gut microbiome exists and how such a spatial organization could be established. In order to pave the way for answering this question, we generated an optimized and adapted fluorescence in situ hybridization (FISH) protocol. We focused on the detection of the two major Drosophila gut microbiome constituting bacteria genera: Acetobacter and Lactobacillus. FISH allows to detect the bacteria in situ and thus to investigate their spatial localization in respect to the host as well as to other microbiome members. We demonstrate the applicability of the protocol using a diverse set of sample types.


Assuntos
Bactérias/genética , DNA Bacteriano/genética , Drosophila melanogaster/microbiologia , Hibridização in Situ Fluorescente/métodos , Acetobacter/genética , Acetobacter/isolamento & purificação , Animais , Bactérias/isolamento & purificação , Fezes/microbiologia , Trato Gastrointestinal/microbiologia , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Masculino , Análise Espacial
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