RESUMO
Habituation is the adaptive behavioral outcome of processes engaged in timely devaluation of non-reinforced repetitive stimuli, but the neuronal circuits and molecular mechanisms that underlie them are not well understood. To gain insights into these processes we developed and characterized a habituation assay to repetitive footshocks in mixed sex Drosophila groups and demonstrated that acute neurotransmission from adult α/ß mushroom body (MB) neurons prevents premature stimulus devaluation. Herein we demonstrate that activity of the non-receptor tyrosine kinase dBtk protein is required within these neurons to prevent premature habituation. Significantly, we also demonstrate that the complementary process of timely habituation to the repetitive stimulation is facilitated by α'/ß' MB neurons and also requires dBtk activity. Hence our results provide initial insights into molecular mechanisms engaged in footshock habituation within distinct MB neurons. Importantly, dBtk attenuation specifically within α'/ß' neurons leads to defective habituation, which is readily reversible by administration of the antipsychotics clozapine and risperidone suggesting that the loss of the kinase may dysregulate monoamine receptors within these neurons, whose activity underlies the failure to habituate.SIGNIFICANCE STATEMENT Habituation refers to processes underlying decisions to attend or ignore stimuli, which are pivotal to brain function as they underlie selective attention and learning, but the circuits involved and the molecular mechanisms engaged by the process therein are poorly understood. We demonstrate that habituation to repetitive footshock involves two phases mediated by distinct neurons of the Drosophila mushroom bodies and require the function of the dBtk non-receptor tyrosine kinase. Moreover, habituation failure upon dBtk abrogation in neurons where it is required to facilitate the process is readily reversible by antipsychotics, providing conceptual links to particular symptoms of schizophrenia in humans, also characterized by habituation defects and ameliorated by these pharmaceuticals.
Assuntos
Tirosina Quinase da Agamaglobulinemia/fisiologia , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Habituação Psicofisiológica/fisiologia , Corpos Pedunculados/fisiologia , Proteínas Tirosina Quinases/metabolismo , Tirosina Quinase da Agamaglobulinemia/genética , Animais , Animais Geneticamente Modificados , Proteínas de Drosophila/genética , Eletrochoque , Feminino , Masculino , Mutação , Transmissão SinápticaRESUMO
BACKGROUND: Statistical analyses in human populations have associated limited food availability during development with increased longevity of next generations. In support, recent findings in Caenorhabditis elegans revealed nutritional effects on transgenerational longevity. OBJECTIVES: In this study we tested the effect of nutrition on longevity of future generations in Drosophila and whether this is sex-specific. METHODS: We reared male larvae and adults of Drosophila under different food conditions and performed lifespan analyses in F2 generation. RESULTS: Grandsons of males which experienced starvation through larval stages were long-lived and grandsons of well fed larvae were short lived, in two Drosophila strains. In one strain, the nutritional effect on transgenerational longevity was transmitted through male line. Interestingly, we find that dietary restriction in adult males is the main nutritional condition affecting lifespan of grandsons. CONCLUSIONS: Our findings suggest that nutritional regulation of transgenerational longevity is evolutionarily conserved and developmental stage - dependent in Drosophila.
RESUMO
Even though in vivo imaging approaches have witnessed several new and important developments, specimens that exhibit high light scattering properties such as Drosophila melanogaster pupae are still not easily accessible with current optical imaging techniques, obtaining images only from subsurface features. This means that in order to obtain 3D volumetric information these specimens need to be studied either after fixation and a chemical clearing process, through an imaging window--thus perturbing physiological development -, or during early stages of development when the scattering contribution is negligible. In this paper we showcase how Optical Projection Tomography may be used to obtain volumetric images of the head eversion process in vivo in Drosophila melanogaster pupae, both in control and headless mutant specimens. Additionally, we demonstrate the use of Helical Optical Projection Tomography (hOPT) as a tool for high throughput 4D-imaging of several specimens simultaneously.
Assuntos
Drosophila melanogaster/fisiologia , Imageamento Tridimensional/métodos , Tomografia Óptica/métodos , Animais , Diagnóstico por Imagem/métodos , Pupa/fisiologiaRESUMO
Certain xenobiotics have the capacity to induce the expression of genes involved in various biological phenomena, including insecticide resistance. The induction potential of different chemicals, among them different insecticides, has been documented for a number of insect species. In this study, we have analyzed the induction potential of Imidacloprid, a widely used member of the neonicotinoid insecticide family. Genes Cyp6g1 and Cyp6a2, known to be involved in the resistance of mutant Drosophila melanogaster line MiT[Wâ»]3R2 to Imidacloprid and DDT were included in the analyzed sample. We find that Imidacloprid does not induce expression of the analyzed genes.
Assuntos
Sistema Enzimático do Citocromo P-450/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/genética , Imidazóis/toxicidade , Nitrocompostos/toxicidade , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Família 6 do Citocromo P450 , DDT/toxicidade , Proteínas de Drosophila/metabolismo , Regulação da Expressão Gênica , Resistência a Inseticidas , Inseticidas/toxicidade , Neonicotinoides , Xenobióticos/toxicidadeRESUMO
Insecticide resistance is a worldwide problem with major impact on agriculture and human health. Understanding the underlying molecular mechanisms is crucial for the management of the phenomenon; however, this information often comes late with respect to the implementation of efficient counter-measures, particularly in the case of metabolism-based resistance mechanisms. We employed a genome-wide insertional mutagenesis screen to Drosophila melanogaster, using a Minos-based construct, and retrieved a line (MiT[w(-)]3R2) resistant to the neonicotinoid insecticide Imidacloprid. Biochemical and bioassay data indicated that resistance was due to increased P450 detoxification. Deep sequencing transcriptomic analysis revealed substantial over- and under-representation of 357 transcripts in the resistant line, including statistically significant changes in mixed function oxidases, peptidases and cuticular proteins. Three P450 genes (Cyp4p2, Cyp6a2 and Cyp6g1) located on the 2R chromosome, are highly up-regulated in mutant flies compared to susceptible Drosophila. One of them (Cyp6g1) has been already described as a major factor for Imidacloprid resistance, which validated the approach. Elevated expression of the Cyp4p2 was not previously documented in Drosophila lines resistant to neonicotinoids. In silico analysis using the Drosophila reference genome failed to detect transcription binding factors or microRNAs associated with the over-expressed Cyp genes. The resistant line did not contain a Minos insertion in its chromosomes, suggesting a hit-and-run event, i.e. an insertion of the transposable element, followed by an excision which caused the mutation. Genetic mapping placed the resistance locus to the right arm of the second chromosome, within a â¼1 Mb region, where the highly up-regulated Cyp6g1 gene is located. The nature of the unknown mutation that causes resistance is discussed on the basis of these results.
Assuntos
Mapeamento Cromossômico/métodos , Drosophila melanogaster/genética , Perfilação da Expressão Gênica/métodos , Resistência a Inseticidas/genética , Mutagênese/genética , Animais , Bioensaio , Cromossomos de Insetos/genética , Biologia Computacional , DDT/toxicidade , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Drosophila melanogaster/efeitos dos fármacos , Feminino , Genes de Insetos/genética , Loci Gênicos/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Imidazóis/toxicidade , Inativação Metabólica/genética , Resistência a Inseticidas/efeitos dos fármacos , Masculino , Anotação de Sequência Molecular , Neonicotinoides , Nitrocompostos/toxicidade , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
The maternally inherited obligatory intracellular bacterium Wolbachia is a reproductive parasite of many insect species. Wolbachia evades the host immune system, uses the mitotic apparatus to ensure infection of daughter cells, migrates through the host to the gonads and causes reproductive phenotypes, most commonly cytoplasmic incompatibility (CI), i.e. incompatibility of sperm from infected males and eggs from uninfected females. Due to the interconnected facts that Wolbachia is not ex vivo culturable and that no established transformation system exists, virtually nothing is known about Wolbachia-host interactions at the macromolecular level. Intriguingly, the Wolbachia genome codes for an unusually high number of ankyrin repeat (ANK) proteins. ANKs mediate protein-protein interactions in many different contexts. More common in eukaryotes, they also occur in prokaryotes. Some intracellular pathogenic bacteria export ANK effector proteins to the host cytoplasm. This makes the Wolbachia ANK genes candidates for mediating interactions with host cells. We quantified expression of ANK genes of Wolbachia strain wMel in adult gonads and detected host sex-specific regulation of two wMel ANK genes in the gonads in two different backgrounds. Regulation was tissue-specific and independent of host background. We further analyzed expression of their homologues in strains wAu and wRi and found regulation only in wAu. Regulation was tissue-specific and there was no correlation between regulation of these genes and the ability of a strain to induce CI.
Assuntos
Drosophila/genética , Drosophila/microbiologia , Regulação da Expressão Gênica , Wolbachia/genética , Animais , Animais Geneticamente Modificados , Repetição de Anquirina , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Drosophila/metabolismo , Feminino , Gônadas/metabolismo , Gônadas/microbiologia , Masculino , Dados de Sequência Molecular , Especificidade de Órgãos , Wolbachia/químicaRESUMO
Wolbachia is an obligatory intracellular bacterium which often manipulates the reproduction of its insect and isopod hosts. In contrast, Wolbachia is an essential symbiont in filarial nematodes. Lately, Wolbachia has been implicated in genomic imprinting of host DNA through cytosine methylation. The importance of DNA methylation in cell fate and biology calls for in depth studying of putative methylation-related genes. We present a molecular and phylogenetic analysis of a putative DNA adenine methyltransferase encoded by a prophage in the Wolbachia genome. Two slightly different copies of the gene, met1 and met2, exhibit a different distribution over various Wolbachia strains. The met2 gene is present in the majority of strains, in wAu, however, it contains a frameshift caused by a 2 bp deletion. Phylogenetic analysis of the met2 DNA sequences suggests a long association of the gene with the Wolbachia host strains. In addition, our analysis provides evidence for previously unnoticed multiple infections, the detection of which is critical for the molecular elucidation of modification and/or rescue mechanism of cytoplasmic incompatibility.
Assuntos
Drosophila/microbiologia , Genes Virais/genética , Interações Hospedeiro-Patógeno/genética , Prófagos/enzimologia , DNA Metiltransferases Sítio Específica (Adenina-Específica)/genética , Wolbachia/virologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Genoma Bacteriano/genética , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Filogenia , Reação em Cadeia da Polimerase , Prófagos/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNA , DNA Metiltransferases Sítio Específica (Adenina-Específica)/química , Wolbachia/genéticaRESUMO
The Drosophila Gene Disruption Project (GDP) has created a public collection of mutant strains containing single transposon insertions associated with different genes. These strains often disrupt gene function directly, allow production of new alleles, and have many other applications for analyzing gene function. Here we describe the addition of â¼7600 new strains, which were selected from >140,000 additional P or piggyBac element integrations and 12,500 newly generated insertions of the Minos transposon. These additions nearly double the size of the collection and increase the number of tagged genes to at least 9440, approximately two-thirds of all annotated protein-coding genes. We also compare the site specificity of the three major transposons used in the project. All three elements insert only rarely within many Polycomb-regulated regions, a property that may contribute to the origin of "transposon-free regions" (TFRs) in metazoan genomes. Within other genomic regions, Minos transposes essentially at random, whereas P or piggyBac elements display distinctive hotspots and coldspots. P elements, as previously shown, have a strong preference for promoters. In contrast, piggyBac site selectivity suggests that it has evolved to reduce deleterious and increase adaptive changes in host gene expression. The propensity of Minos to integrate broadly makes possible a hybrid finishing strategy for the project that will bring >95% of Drosophila genes under experimental control within their native genomic contexts.
Assuntos
Elementos de DNA Transponíveis , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Genes de Insetos , Mutagênese Insercional/métodos , Alelos , Animais , Expressão Gênica , Genoma de Inseto , Modelos Genéticos , Mutação , FenótipoRESUMO
Transposons are powerful tools for conducting genetic manipulation and functional studies in organisms that are of scientific, economic, or medical interest. Minos, a member of the Tc1/mariner family of DNA transposons, exhibits a low insertional bias and transposes with high frequency in vertebrates and invertebrates. Its use as a tool for transgenesis and genome analysis of rather different animal species is described.
Assuntos
Elementos de DNA Transponíveis/genética , Técnicas de Transferência de Genes , Genômica/métodos , Invertebrados/genética , Vertebrados/genética , AnimaisRESUMO
Developments in insect transgenesis using transposons combined with available mass rearing technology for insects such as the Medfly, Ceratitis capitata, provide opportunity for the production of protein for industrial, agricultural and healthcare purposes on a very large scale. In this study, we report the germ-line transformation and expression of a cDNA encoding human growth hormone (hGH) in transgenic Drosophila using the Minos transposon. Production and secretion of a bioactive hGH into the haemolymph of transgenic larvae was demonstrated by immunoblot analysis, ELISA and a proliferation bioassay. Stable expression of hGH was observed over 50 generations. The results indicate that mass reared transgenic diptera with a rapid period of larval growth could provide cost effective production systems for the manufacture of therapeutic and other high value proteins.
Assuntos
Drosophila melanogaster/genética , Hormônio do Crescimento Humano/genética , Animais , Animais Geneticamente Modificados , Sequência de Bases , Primers do DNA/genética , Elementos de DNA Transponíveis/genética , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Hemolinfa/metabolismo , Hormônio do Crescimento Humano/biossíntese , Humanos , Larva/efeitos dos fármacos , Larva/genética , Larva/metabolismo , Masculino , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Tetraciclina/farmacologiaRESUMO
Approximately 30 kb of genomic DNA enclosing the Adh locus from the medfly, Ceratitis capitata have been cloned and about 15 kb has been structurally and functionally characterized. The locus consists of two genes, Adh-1 and Adh-2, separated by an intergenic region, which is polymorphic in size ranging from approximately 6.4 kb to 8.1 kb. Both genes consist of three exons and two introns. The introns are below 200 bp in size, except the 1st intron of Adh-1, which is unexpectedly long, variable in size and contains a deleted mariner-like element (postdoc). The two genes are transcribed in different orientations. The Adh-2 gene shows the typical pattern of transcription seen in the homologous genes of Drosophilidae presenting high levels of expression in the fat body, gut and ovaries. The Adh-1 gene is only expressed in the body muscle tissues of embryos, larvae and adult flies, raising the question of what its biological function may be. A DNA fragment containing bases -102 to -1666 relative to the first base of the initiating ATG of Adh-1 is sufficient to drive the expression of a reporter gene in body muscles of Drosophila melanogaster embryos, larvae and adult flies. The study provides further insights into the evolution of the Adh genes of higher diptera.
Assuntos
Álcool Desidrogenase/genética , Ceratitis capitata/genética , Genes de Insetos , Animais , Evolução Biológica , Ceratitis capitata/crescimento & desenvolvimento , DNA Intergênico , Drosophila melanogaster/genética , Feminino , Duplicação Gênica , Expressão Gênica , Genes Reporter , Masculino , Dados de Sequência Molecular , Músculos/metabolismo , Análise de Sequência de DNARESUMO
Much of the information about the function of D. melanogaster genes has come from P-element mutagenesis. The major drawback of the P element, however, is its strong bias for insertion into some genes (hotspots) and against insertion into others (coldspots). Within genes, 5'-UTRs are preferential targets. For the successful completion of the Drosophila Genome Disruption Project, the use of transposon vectors other than P will be necessary. We examined here the suitability of the Minos element from Drosophila hydei as a tool for Drosophila genomics. Previous work has shown that Minos, a member of the Tc1/mariner family of transposable elements, is active in diverse organisms and cultured cells; it produces stable integrants in the germ line of several insect species, in the mouse, and in human cells. We generated and analyzed 96 Minos integrations into the Drosophila genome and devised an efficient "jump-starting" scheme for production of single insertions. The ratio of insertions into genes vs. intergenic DNA is consistent with a random distribution. Within genes, there is a statistically significant preference for insertion into introns rather than into exons. About 30% of all insertions were in introns and approximately 55% of insertions were into or next to genes that have so far not been hit by the P element. The insertion sites exhibit, in contrast to other transposons, little sequence requirement beyond the TA dinucleotide insertion target. We further demonstrate that induced remobilization of Minos insertions can delete nearby sequences. Our results suggest that Minos is a useful tool complementing the P element for insertional mutagenesis and genomic analysis in Drosophila.
Assuntos
Elementos de DNA Transponíveis/genética , Drosophila melanogaster/genética , Genômica/métodos , Mutagênese Insercional/métodos , Transposases/genética , Animais , Sequência de Bases , Primers do DNA , Ligação de Hidrogênio , Dados de Sequência Molecular , Análise de Sequência de DNARESUMO
The first intron of the gene encoding one of the alcohol dehydrogenase isoenzymes (ADH-1) in Ceratitis capitata is highly polymorphic in size. Five size variants of this intron were isolated from different strains and populations and characterized. Restriction map and sequence analysis showed that the intron size polymorphism is due to the presence or absence of (a) a copy of a defective mariner-like element, postdoc; (b) an approximately 550-bp 3' indel which exhibits no similarity to any known sequence; and (c) a central duplication of 704 bp consisting of part of the 3' end of the postdoc element, the region between postdoc and the 3' indel, and the first 20 bp of the 3' indel. The homologous Adh-1 intron was amplified from the congeneric species, Ceratitis rosa, in order to obtain an outgroup for comparative and phylogenetic analyses. The C. rosa introns were polymorphic in size, ranging from about 1100 to 2000 bp, the major difference between them being the presence or absence of a mariner-like element Crmar2, unrelated to the postdoc element. Phylogenetic analysis suggests that the shorter intron variants in C. capitata may represent the ancestral form of the intron, the longest variants apparently being the most recent.
Assuntos
Álcool Desidrogenase/genética , Ceratitis capitata/genética , Proteínas de Insetos/genética , Íntrons/genética , Polimorfismo Genético , Animais , Composição de Bases , Sequência de Bases , Eletroforese em Gel de Ágar , Duplicação Gênica , Funções Verossimilhança , Modelos Genéticos , Dados de Sequência Molecular , Filogenia , Mapeamento por Restrição , Análise de Sequência de DNA , Homologia de SequênciaRESUMO
Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations. Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents. Wolbachia manipulate host reproduction by using several strategies, one of which is cytoplasmic incompatibility (CI) [Stouthamer, R., Breeuwer, J. A. J. & Hurst, G. D. D. (1999) Annu. Rev. Microbiol. 53, 71-102]. We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor. Wolbachia induced complete CI in the novel host. Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced CI could be used as a novel environmentally friendly tool for the control of medfly populations. The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations.
Assuntos
Ceratitis capitata/microbiologia , Controle Biológico de Vetores/métodos , Wolbachia/fisiologia , Animais , Citoplasma/microbiologia , Feminino , Masculino , Tephritidae/microbiologia , Wolbachia/patogenicidadeRESUMO
Wolbachia strains are endosymbiotic bacteria typically found in the reproductive tracts of arthropods. These bacteria manipulate host reproduction to ensure maternal transmission. They are usually transmitted vertically, so it has been predicted that they have evolved a mechanism to target the host's germ cells during development. Through cytological analysis we found that Wolbachia strains display various affinities for the germ line of Drosophila. Different Wolbachia strains show posterior, anterior, or cortical localization in Drosophila embryos, and this localization is congruent with the classification of the organisms based on the wsp (Wolbachia surface protein) gene sequence. This embryonic distribution pattern is established during early oogenesis and does not change until late stages of embryogenesis. The posterior and anterior localization of Wolbachia resembles that of oskar and bicoid mRNAs, respectively, which define the anterior-posterior axis in the Drosophila oocyte. By comparing the properties of a single Wolbachia strain in different host backgrounds and the properties of different Wolbachia strains in the same host background, we concluded that bacterial factors determine distribution, while bacterial density seems to be limited by the host. Possible implications concerning cytoplasmic incompatibility and evolution of strains are discussed.
Assuntos
Drosophila/microbiologia , Interações Hospedeiro-Parasita , Wolbachia/patogenicidade , Animais , Blastoderma/microbiologia , Drosophila/embriologia , Embrião não Mamífero/microbiologia , Feminino , Morfogênese , Ovário/microbiologia , Especificidade da EspécieRESUMO
A system for population control of insects is proposed. It is based on transgenic insects expressing an enzyme which converts an inactive pro-drug into an active, toxic form. A model system is presented which relies on transposon-mediated integration of a bacterial cytosine deaminase (CD) gene into the genome of Drosophila melanogaster. We demonstrate female-specific sterility and transgene-dependent lethality when flies carrying the CD gene under a Drosophila female-specific promoter/enhancer are treated with 5-Fluorocytosine, a low-toxicity nucleoside analogue which is converted to toxic 5-Fluorouracil by the enzyme. The approach can be used with existing pro-insecticides and appropriate converting enzymes in combination with established mass rearing technology, for targeted, environmentally acceptable control of insects of economic and public health importance.
Assuntos
Drosophila melanogaster/genética , Infertilidade Feminina/enzimologia , Infertilidade Feminina/genética , Controle Biológico de Vetores/métodos , Pró-Fármacos/farmacocinética , Animais , Animais Geneticamente Modificados/genética , Citosina Desaminase/genética , Citosina Desaminase/metabolismo , Elementos de DNA Transponíveis/genética , Drosophila melanogaster/enzimologia , Escherichia coli/enzimologia , Escherichia coli/genética , Feminino , Flucitosina/farmacocinética , Fluoruracila/farmacocinética , Genes de Insetos , Homozigoto , Masculino , Pró-Fármacos/farmacologia , Regiões Promotoras Genéticas/genéticaRESUMO
This review summarizes structural and functional studies on medfly promoters and regulatory elements that can be used for driving sex-specific, conditional and constitutive gene expression in this species. Sex-specific and conditional promoters are important for generating transgenic sexing strains that could increase the performance of the Sterile Insect Technique while strong constitutive promoters are necessary for developing sensitive transgenic marker systems. The review focuses on the functional analysis of the promoters of two male-specific and heat shock medfly genes. A special emphasis is put on the potential utility of these promoters for developing transgenic sexing strains.
Assuntos
Ceratitis capitata/genética , Infertilidade/genética , Controle Biológico de Vetores/métodos , Ceratitis capitata/fisiologia , Cloranfenicol O-Acetiltransferase/genética , Cloranfenicol O-Acetiltransferase/metabolismo , Proteínas de Choque Térmico HSP70/genética , Regiões Promotoras Genéticas/genética , Pré-Seleção do Sexo/métodos , Transformação GenéticaRESUMO
We report the first systematic survey for the presence of Wolbachia endosymbionts in aphids and whiteflies, particularly different populations and biotypes of Bemisia tabaci. Additional agriculturally important species included were predator species, leafhoppers, and lepidopterans. We used a polymerase chain reaction (PCR)-based detection assay with ribosomal 16S rDNA and Wolbachia cell surface protein (wsp) gene primers. Wolbachia were detected in a number of whitefly populations and species, whitefly predators, and one leafhopper species; however, none of the aphid species tested were found infected. Single, double, and triple infections were detected in some of the B. tabaci populations. PCR and phylogenetic analysis of wsp gene sequences indicated that all Wolbachia strains found belong to group B. Topologies of the optimal tree derived by maximum likelihood (ML) and a ML tree in which Wolbachia sequences from B. tabaci are constrained to be monophyletic are significantly different. Our results indicate that there have been at least four independent Wolbachia infection events in B. tabaci. The importance of the presence of Wolbachia infections in B. tabaci is discussed.
Assuntos
Afídeos/microbiologia , Hemípteros/microbiologia , Wolbachia/isolamento & purificação , Animais , Proteínas da Membrana Bacteriana Externa/genética , DNA Bacteriano/análise , DNA Ribossômico/análise , Insetos/microbiologia , Lepidópteros/microbiologia , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Simbiose , Wolbachia/classificação , Wolbachia/genéticaRESUMO
Wolbachia are a group of maternally transmitted obligatory intracellular alpha-proteobacteria that infect a wide range of arthropod and nematode species. Wolbachia infection in Drosophila in most cases is associated with the induction of cytoplasmic incompatibility (CI), manifested as embryonic lethality of offspring in a cross between infected males and uninfected females. While the molecular basis of CI is still unknown, it has been suggested that two bacterial functions are involved: mod (for modification) modifies the sperm during spermatogenesis and resc (for rescue) acts in the female germline and/or in early embryos, neutralizing the modification. There is considerable variation in the level of incompatibility in different Wolbachia/host interactions. We examine the relationship between the levels of CI in a number of naturally infected and transinfected Drosophila hosts and the percentage of Wolbachia-infected sperm cysts. Our results indicate the presence of two main groups of Drosophila-Wolbachia associations: group I, which exhibits a positive correlation between CI levels and the percentage of infected sperm cysts (mod(+) phenotype), and group II, which does not express CI (mod(-) phenotype) irrespective of the infection status of the sperm cysts. Group II can be further divided into two subgroups: The first one contains associations with high numbers of heavily Wolbachia-infected sperm cysts while in the second one, Wolbachia is rarely detected in sperm cysts, being mostly present in somatic cells. We conclude that there are three requirements for the expression of CI in a host-Wolbachia association: (a) Wolbachia has to be able to modify sperm (mod(+) genotype), (b) Wolbachia has to infect sperm cysts, and (c) Wolbachia has to be harbored by a permissive host.
Assuntos
Drosophila melanogaster/microbiologia , Wolbachia/patogenicidade , Animais , Cruzamentos Genéticos , Cistos/microbiologia , Citoplasma/metabolismo , Feminino , Genótipo , Masculino , Microscopia Confocal , Fenótipo , Espermatogênese , Espermatozoides/metabolismo , Temperatura , Testículo/microbiologiaRESUMO
We tested the suitability of the fly transposon Minos, a member of the Tc1/mariner superfamily, for insertional mutagenesis in the mouse germ line. We generated a transgenic mouse line expressing Minos transposase in growing oocytes and another carrying a tandem array of nonautonomous transposons. The frequency of transposition in the progeny derived from oocytes carrying both transgenes is 8.2%. Analysis of the new integration sites shows a high frequency of transpositions to a different chromosome. Thus Minos transposition could be an effective system for insertional mutagenesis and functional genomic analysis in the mouse.