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1.
Sci Rep ; 12(1): 219, 2022 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-34997157

RESUMO

A germ-free rearing system is a crucial method for host-microbiota interactions using Nasonia as a model system. The previous rearing media in 2012 introduced toxic factors like bleach and antibiotics, required significant effort and volume of media preparation, and the rearing protocols in 2012 and 2016 often resulted in embryos, larvae, and enclosing pupae drowning, underfed, or desiccating. In this work, we optimize the germ-free rearing media that excludes the toxic factors and provide a substrate for the developing animals to have constant access to media without the risk of drowning or desiccation. The new process resulted in an increase in full maturation of larvae to adults from 33 to 65%, with no effect on the rate of growth or final adult size. This significantly improves the applicability of germ-free rearing of Nasonia and potentially other parasitoids.


Assuntos
Dípteros/crescimento & desenvolvimento , Entomologia/métodos , Animais , Feminino , Vida Livre de Germes , Interações entre Hospedeiro e Microrganismos , Larva/crescimento & desenvolvimento , Masculino , Pupa/crescimento & desenvolvimento
2.
Sci Rep ; 12(1): 1105, 2022 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-35058520

RESUMO

Magnetic resonance imaging (MRI) is the key whole-body imaging technology for observing processes within a living object providing excellent resolution and contrast between soft tissues. In the present work, we exploited the non-destructive properties of MRI to track longitudinally the dynamic changes that take place in developing pupae of the Emperor Moth (Saturnia pavonia) during the last days before eclosion. While in diapause pupae, body fluid was almost homogeneously distributed over the internal compartments, as soon as wings, legs, flight muscles and the head region were fully developed, a significant redistribution of water levels occurred between thoracic and abdominal regions. During the last two days before eclosion, the developing moths transferred substantial amounts of liquid into the gut and the labial gland, and in case of females, into developing eggs. Concomitantly, the volume of the air sacs increased drastically and their expansion/compression became clearly visible in time-resolved MR images. Furthermore, besides ventilation of the tracheal system, air sacs are likely to serve as volume reservoir for liquid transfer during development of the moths inside their pupal case. In parallel, we were able to monitor noninvasively lipid consumption, cardiac activity and haemolymph circulation during final metamorphosis.


Assuntos
Lepidópteros/crescimento & desenvolvimento , Metamorfose Biológica/fisiologia , Pupa/crescimento & desenvolvimento , Animais , Lepidópteros/metabolismo , Imageamento por Ressonância Magnética/métodos , Mariposas/fisiologia
3.
J Med Entomol ; 59(1): 147-161, 2022 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-34698350

RESUMO

Studies under constant temperatures are the most common to estimate the Postmortem Interval (PMI). It is imperative that forensic sciences have data from studies carried out in the field. Therefore, this work aims to: (1) evaluate the parameters (weight, length, development time) associated with the life cycles of Lucilia ochricornis (Wiedemann) (Diptera: Calliphoridae) and Lucilia purpurascens (Walker) under experimental conditions in the field considering fluctuating temperatures, and (2) compare these results with those known and published by the same authors for cultures realized in the laboratory under constant temperatures; which will permit us to contrast the most widely used existing methodologies for forensic application in estimating the minimum postmortem interval (PMImin). For each season of the year, cultures of both species were made in the field, collecting information on temperature, humidity, and photoperiod to perform laboratory cultures, later comparing: development time, length, weight, and Accumulated Degree-Hours (ADH) in both types of cultures. Methods for estimating the PMI were obtained and validated with the information of the cultures grown in the field. The two types of cultures showed differences between each other for both species. The forensic use methods to estimate PMI were enhanced and their precision increased when maximum larval length data were used, and it was also concluded that feeding larval stages are the most accurate to be used in making estimates because the larva is growing. The estimation of the PMI through the use of necrophagous flies development remains reliable for obtaining the PMImin.


Assuntos
Calliphoridae/fisiologia , Traços de História de Vida , Animais , Argentina , Calliphoridae/crescimento & desenvolvimento , Temperatura Baixa , Feminino , Entomologia Forense , Temperatura Alta , Larva/crescimento & desenvolvimento , Larva/fisiologia , Masculino , Pupa/crescimento & desenvolvimento , Pupa/fisiologia , Estações do Ano , Especificidade da Espécie , Temperatura
4.
J Med Entomol ; 59(1): 173-183, 2022 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-34661674

RESUMO

The family Culicidae is represented by 244 species in Argentina, many of them with epidemiological importance. DNA barcodes are effective tools for identifying mosquito species, for knowing genetic variability, and for establishing phylogenetic relationships. This work aims to explore mosquito diversity employing different species delimitation approaches and to establish formally a DNA barcode library for the Argentinian mosquito fauna. Barcode fragments of 80 specimens of Argentinian mosquitoes of 28 species of the genera Aedeomyia Theobald (Diptera: Culicidae), Anopheles Meigen (Diptera: Culicidae), Coquillettidia Dyar (Diptera: Culicidae), Culex L. (Diptera: Culicidae), Haemagogus Williston (Diptera: Culicidae), Mansonia Blanchard (Diptera: Culicidae), Nyssorhynchus Blanchard (Diptera: Culicidae), Ochlerotatus Lynch-Arribálzaga (Diptera: Culicidae), Psorophora Robinneau-Desvoidy (Diptera: Culicidae) and Uranotaenia Lynch-Arribálzaga (Diptera: Culicidae) were sequenced. Another 82 sequences were obtained from public databases to establish the phylogenetic relationships using Maximum Likelihood and Bayesian Inference, and the species boundaries based on three approaches (ABGD, GMYC, and mPTP). Sixteen of the 28 species sequenced were recovered as monophyletic, of which 12 were also recognized as molecular operational taxonomic units according to the three methodologies. The disparity between morphology and barcode-based identifications could be explained by synonymy, species complexes occurrence, hybridization, incomplete lineage sorting, or the effect of the geographical scale of sampling. Twenty of the 28 sequenced species are new barcodes for Argentina and 11 are the first for science. This increases from 31 to 52 (12.7 to 21.31%) and from six to 10 (28.57 to 47.62%) the number of species and genera, respectively, with barcode sequences in Argentina. New species records are provided.


Assuntos
Distribuição Animal , Culicidae/classificação , Código de Barras de DNA Taxonômico , Animais , Argentina , Culicidae/crescimento & desenvolvimento , Feminino , Larva/classificação , Larva/crescimento & desenvolvimento , Masculino , Filogenia , Pupa/classificação , Pupa/crescimento & desenvolvimento
5.
J Med Entomol ; 59(1): 108-119, 2022 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-34668022

RESUMO

Sarcophaga peregrina (Robineau-Desvoidy, 1830), a synanthropic flesh fly species found in different parts of the world, is of medical and forensic importance. Traditional methods of inferring developmental age rely on the life stage of insects and morphological changes. However, once the larvae reach the pupal and adult stage, morphological changes would become barely visible, so that the classic method would be invalid. Here, we studied the cuticular hydrocarbon profile of S. peregrina of the whole life cycle from larval stage to adult stage by GC-MS. Sixty-three compounds with carbon chain length ranging from 8 to 36 were detected, which could be categorized into four classes: n-alkanes, branched alkanes, alkenes, and unknowns. As developmental increased, branched alkanes dominant, and the content of high-molecular-weight hydrocarbons is variable, especially for 2-methyl C19, DiMethyl C21, docosane (C22), and tricosane (C23). This study shows that the composition of CHC could be used to determine the developmental age of S. peregrina and aid in postmortem interval estimations in forensic science.


Assuntos
Hidrocarbonetos/metabolismo , Sarcofagídeos/química , Animais , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Larva/química , Larva/crescimento & desenvolvimento , Masculino , Óvulo/química , Pupa/química , Pupa/crescimento & desenvolvimento
6.
Infect Genet Evol ; 97: 105185, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34920099

RESUMO

Aedes aegypti is a vector of several global human viruses responsible for high human morbidity and mortality. The method to prevent the transmission of vector-borne viruses is mainly based on the control of the insect vector using insecticides. Among these chemicals, copper sulfate is a compound widely used in agriculture with the potential to be used as an alternative to control these insects. This study evaluated the effects of the exposure of A. aegypti larvae to copper sulfate on survival, midgut morphology, blood-feeding and fecundity. The exposure to CuSO4 decreased the survival of A. aegypti during the immature phase. Adults obtained from exposed larvae had their lifespan decreased at all tested concentrations. The exposure to CuSO4 impaired the development in the transition from larvae to pupae and from pupae to adult. The number of eggs laid by females developed from larvae treated with CuSO4 was significantly lower than in control. In addition, the egg hatching rates were also negatively affected. The midguts of treated larvae and pupae showed epithelial disorganization. The number of cleaved caspase-3 cells increased in the midgut of exposed pupae compared to control. Moreover, there was a reduction in proliferating cells in treated larvae and pupae compared to the control. In conclusion, the results reveal that CuSO4 exposure has insecticidal activity against A. aegypti, which may be related to the impairment of the midgut metamorphosis and reduced proliferation of stem cells, with the consequent impairment of female mosquito fertility and fecundity.


Assuntos
Aedes , Sulfato de Cobre , Inseticidas , Aedes/crescimento & desenvolvimento , Animais , Sistema Digestório/efeitos dos fármacos , Sistema Digestório/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Longevidade/efeitos dos fármacos , Pupa/crescimento & desenvolvimento , Reprodução/efeitos dos fármacos
7.
Development ; 148(23)2021 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-34861038

RESUMO

Within developing tissues, cell proliferation, cell motility and other cell behaviors vary spatially, and this variability gives a complexity to the morphogenesis. Recently, novel formalisms have been developed to quantify tissue deformation and underlying cellular processes. A major challenge for the study of morphogenesis now is to objectively define tissue sub-regions exhibiting different dynamics. Here, we propose a method to automatically divide a tissue into regions where the local deformation rate is homogeneous. This was achieved by several steps including image segmentation, clustering and region boundary smoothing. We illustrate the use of the pipeline using a large dataset obtained during the metamorphosis of the Drosophila pupal notum. We also adapt it to determine regions in which the time evolution of the local deformation rate is homogeneous. Finally, we generalize its use to find homogeneous regions for cellular processes such as cell division, cell rearrangement, or cell size and shape changes. We also illustrate it on wing blade morphogenesis. This pipeline will contribute substantially to the analysis of complex tissue shaping, and the biochemical and biomechanical regulations driving tissue morphogenesis.


Assuntos
Metamorfose Biológica , Modelos Biológicos , Animais , Drosophila melanogaster , Pupa/crescimento & desenvolvimento
8.
mBio ; 12(6): e0296621, 2021 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-34933445

RESUMO

Microbiomes provide a range of benefits to their hosts which can lead to the coevolution of a joint ecological niche. However, holometabolous insects, some of the most successful organisms on Earth, occupy different niches throughout development, with larvae and adults being physiologically and morphologically highly distinct. Furthermore, transition between the stages usually involves the loss of the gut microbiome since the gut is remodeled during pupation. Most eusocial organisms appear to have evolved a workaround to this problem by sharing their communal microbiome across generations. However, whether this vertical microbiome transmission can overcome perturbations of the larval microbiome remains untested. Honey bees have a relatively simple, conserved, coevolved adult microbiome which is socially transmitted and affects many aspects of their biology. In contrast, larval microbiomes are more variable, with less clear roles. Here, we manipulated the gut microbiome of in vitro-reared larvae, and after pupation of the larvae, we inoculated the emerged bees with adult microbiome to test whether adult and larval microbiome stages may be coupled (e.g., through immune priming). Larval treatments differed in bacterial composition and abundance, depending on diet, which also drove larval gene expression. Nonetheless, adults converged on the typical core taxa and showed limited gene expression variation. This work demonstrates that honey bee adult and larval stages are effectively microbiologically decoupled, and the core adult microbiome is remarkably stable to early developmental perturbations. Combined with the transmission of the microbiome in early adulthood, this allows the formation of long-term host-microbiome associations. IMPORTANCE This work investigated host-microbiome interactions during a crucial developmental stage-the transition from larvae to adults, which is a challenge to both, the insect host and its microbiome. Using the honey bee as a tractable model system, we showed that microbiome transfer after emergence overrides any variation in the larvae, indicating that larval and adult microbiome stages are effectively decoupled. Together with the reliable vertical transfer in the eusocial system, this decoupling ensures that the adults are colonized with a consistent and derived microbiome after eclosion. Taken all together, our data provide additional support that the evolution of sociality, at least in the honey bee system tested here, is linked with host-microbiome relationships.


Assuntos
Abelhas/microbiologia , Microbioma Gastrointestinal , Larva/microbiologia , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Abelhas/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Pupa/crescimento & desenvolvimento , Pupa/microbiologia , Reprodução
9.
Cell Rep ; 37(13): 110151, 2021 12 28.
Artigo em Inglês | MEDLINE | ID: mdl-34965423

RESUMO

Ran's GTPase-activating protein (RanGAP) is tethered to the nuclear envelope (NE) in multicellular organisms. We investigated the consequences of RanGAP localization in human tissue culture cells and Drosophila. In tissue culture cells, disruption of RanGAP1 NE localization surprisingly has neither obvious impacts on viability nor nucleocytoplasmic transport of a model substrate. In Drosophila, we identified a region within nucleoporin dmRanBP2 required for direct tethering of dmRanGAP to the NE. A dmRanBP2 mutant lacking this region shows no apparent growth defects during larval stages but arrests at the early pupal stage. A direct fusion of dmRanGAP to the dmRanBP2 mutant rescues this arrest, indicating that dmRanGAP recruitment to dmRanBP2 per se is necessary for the pupal ecdysis sequence. Our results indicate that while the NE localization of RanGAP is widely conserved in multicellular organisms, the targeting mechanisms are not. Further, we find a requirement for this localization during pupal development.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Proteínas Ativadoras de GTPase/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Chaperonas Moleculares/metabolismo , Membrana Nuclear/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Poro Nuclear/metabolismo , Pupa/crescimento & desenvolvimento , Transporte Ativo do Núcleo Celular , Animais , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Proteínas Ativadoras de GTPase/genética , Células HCT116 , Humanos , Chaperonas Moleculares/genética , Membrana Nuclear/genética , Poro Nuclear/genética , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Pupa/genética , Pupa/metabolismo
10.
PLoS Genet ; 17(11): e1009916, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34843450

RESUMO

Insect metamorphosis is triggered by the production, secretion and degradation of 20-hydroxyecdysone (ecdysone). In addition to its role in developmental regulation, increasing evidence suggests that ecdysone is involved in innate immunity processes, such as phagocytosis and the induction of antimicrobial peptide (AMP) production. AMP regulation includes systemic responses as well as local responses at surface epithelia that contact with the external environment. At pupariation, Drosophila melanogaster increases dramatically the expression of three AMP genes, drosomycin (drs), drosomycin-like 2 (drsl2) and drosomycin-like 5 (drsl5). We show that the systemic action of drs at pupariation is dependent on ecdysone signalling in the fat body and operates via the ecdysone downstream target, Broad. In parallel, ecdysone also regulates local responses, specifically through the activation of drsl2 expression in the gut. Finally, we confirm the relevance of this ecdysone dependent AMP expression for the control of bacterial load by showing that flies lacking drs expression in the fat body have higher bacterial persistence over metamorphosis. In contrast, local responses may be redundant with the systemic effect of drs since reduction of ecdysone signalling or of drsl2 expression has no measurable negative effect on bacterial load control in the pupa. Together, our data emphasize the importance of the association between ecdysone signalling and immunity using in vivo studies and establish a new role for ecdysone at pupariation, which impacts developmental success by regulating the immune system in a stage-dependent manner. We speculate that this co-option of immune effectors by the hormonal system may constitute an anticipatory mechanism to control bacterial numbers in the pupa, at the core of metamorphosis evolution.


Assuntos
Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Metamorfose Biológica/genética , Animais , Drosophila melanogaster/crescimento & desenvolvimento , Ecdisona/genética , Ecdisterona/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Larva/genética , Larva/crescimento & desenvolvimento , Pupa/genética , Pupa/crescimento & desenvolvimento , Transdução de Sinais/genética
11.
PLoS One ; 16(10): e0259115, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34699561

RESUMO

Blue-green algae, Spirulina platensis is a well-known algal formulation known for its beneficial effects on the growth and development in several types of organisms. Although it is used as a food supplement, it possesses significant toxic effects on growth and development of organisms. This study assessed the positive/negative impacts of S. platensis on almond moth, Cadra cautella (almond moth) that is a serious pest of date fruits and other grains under laboratory conditions. The S. platensis powder were mixed with diet and newly hatched C. cautella larvae were fed. The larvae were observed on alternate days to record the data. The diet was changed once a week. The S. platensis proved very good nutrition supplement at lower dose. Whereas, moderate and high mortality was noted for 5 and 10% formulations, respectively. Moreover, larval span was significantly altered by different formulations and lower formulation (1%) resulted in shorter larval period compared to the rest of the formulations. Although 33% mortality was recorded under 5% S. platensis formulation, however, the larvae which reached to adult stage, copulated, and females laid more eggs. Furthermore, the highest mortality (90%) was observed under 10% S. platensis formulation and a few larvae reached adult stage; thus, no data on pupal period and reproductive traits was recorded for this formulation. These findings proved that S. platensis can be used as nutritional supplement as well as a toxic substance to manage C. cautella in date storage. However, future studies on this are needed to reach concrete conclusions.


Assuntos
Suplementos Nutricionais , Mariposas/crescimento & desenvolvimento , Phoeniceae/parasitologia , Spirulina , Animais , Larva/crescimento & desenvolvimento , Pupa/crescimento & desenvolvimento
12.
J Insect Sci ; 21(5)2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34636890

RESUMO

Hexamerins are members of the hemocyanin superfamily and play essential roles in providing amino acids and energy for the nonfeeding stages of insects. In this study, we cloned and analyzed the expression patterns of four hexamerin genes (hex 70a, hex 70b, hex 70c, and hex 110) at different worker development stages and queen diapause statuses in the bumble bee, Bombus terrestris. The results of this study showed that hex 110 has the longest open reading frame (ORF; 3,297 bp) compared to the ORFs of hex 70a (2,034 bp), hex 70b (2,067 bp), and hex 70c (2,055 bp). The putative translation product of Hex 70a, Hex 70b, Hex70c, and Hex 110 has 677, 688, 684, and 1,098aa with predicted molecular mass of 81.13, 79.69, 81.58, and 119 kDa. In the development stages of workers, the expression levels of hex 70a, hex 70b, and hex 70c increased gradually from the larval stage and exhibited high expression levels at the pink eyed and brown eyed pupae stage, whereas hex 110 exhibited the highest expression level at the larval period. Four hexamerin genes were highly expressed at the prediapause status of queen (P < 0.05), and compared to the eclosion queen, the lowest upregulation was 3.7-fold, and the highest upregulation was 1,742-fold. The expression levels of hex 70b, hex 70c, and hex 110 at diapause were significantly higher than those at postdiapause (P < 0.05). In conclusion, hexamerins may play important roles in queen diapause and metamorphosis of larval and pupal stages.


Assuntos
Abelhas , Proteínas de Insetos/genética , Animais , Abelhas/genética , Abelhas/crescimento & desenvolvimento , Abelhas/fisiologia , Diapausa/genética , Diapausa/fisiologia , Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Larva/crescimento & desenvolvimento , Larva/fisiologia , Metamorfose Biológica/genética , Metamorfose Biológica/fisiologia , Pupa/genética , Pupa/crescimento & desenvolvimento , Pupa/fisiologia
13.
J Invertebr Pathol ; 186: 107674, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34606828

RESUMO

The insect cuticle is a composite structure that can further be divided into a few sub-structural layers. Its large moiety comprises a lattice of chitin fibrils and structural proteins, both of which are stabilized by covalent bonding among them. The cuticle covers the whole surface of insect body, and thus has long been suggested for the involvement in defense against entomopathogens, especially entomopathogenic fungi that infect percutaneously. We have been addressing this issue in the past few years and have so far demonstrated experimentally that chitin synthase 1, laccase2 as well as benzoquinone synthesis-related genes of Tribolium castaneum have indispensable roles in the antifungal host defense. In the present study we focused on another major component of the insect cuticular integument, structural cuticular proteins. We chose three genes coding for adult-specific cuticular proteins, namely CPR4, CPR18 and CPR27, and examined their roles in forming immunologically sound adult cuticular integuments. Analyses of developmental expression revealed that the three genes showed high level expression in the pupal stage. These results are consistent with their proposed roles in constructing cuticle of adult beetles. The RNA interference-mediated gene knockdown was employed to silence these genes, and the administration of double strand RNAs in pupae resulted in the adults with malformed elytra. The single knockdown of the three genes attenuated somewhat the defense of the resulting adult beetles against Beauveria bassiana and Metarhizium anisopliae, but statistical analyses indicated no significant differences from controls. In contrast, the double or triple knockdown mutant beetles displayed a drastic disruption of the host defense against the two entomopathogenic fungal species irrespective of the combination of targeted cuticular protein genes, demonstrating the important roles of the three cuticular protein genes in conferring robust antifungal properties on the adult cuticle. Scanning electron microscopic observation revealed that the germination of conidia attached on the adult body surface was still suppressed after the gene knockdown as in the case of wild-type beetles, suggesting that the weakened antifungal phenotypes resulted from the combined knockdown of the adult-specific cuticular protein genes could not be accounted for by the disfunction of secretion/retention of fungistatic benzoquinone derivatives.


Assuntos
Beauveria/fisiologia , Proteínas de Insetos/genética , Metarhizium/fisiologia , Tribolium/genética , Animais , Proteínas de Insetos/metabolismo , Pupa/genética , Pupa/crescimento & desenvolvimento , Pupa/metabolismo , Pupa/microbiologia , Tribolium/crescimento & desenvolvimento , Tribolium/metabolismo , Tribolium/microbiologia
14.
Int J Legal Med ; 135(6): 2625-2635, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34570269

RESUMO

Estimating the age of the developmental stages of the blow fly Calliphora vicina (Diptera: Calliphoridae) is of forensic relevance for the determination of the minimum post-mortem interval (PMImin). Fly eggs and larvae can be aged using anatomical and morphological characters and their modification during development. However, such methods can only hardly be applied for aging fly pupae. Previous study described age estimation of C. vicina pupae using gene expression, but just when reared at constant temperatures, but fluctuating temperatures represent a more realistic scenario at a crime scene. Therefore, age-dependent gene expression of C. vicina pupae were compared at 3 fluctuating and 3 constant temperatures, the latter representing the mean values of the fluctuating profiles. The chosen marker genes showed uniform expression patterns during metamorphosis of C. vicina pupae bred at different temperature conditions (constant or fluctuating) but the same mean temperature (e.g. constant 10 °C vs. fluctuating 5-15 °C). We present an R-based statistical tool, which enables estimation of the age of the examined pupa based on the analysed gene expression data.


Assuntos
Calliphoridae/crescimento & desenvolvimento , Calliphoridae/genética , Expressão Gênica , Metamorfose Biológica , Pupa/crescimento & desenvolvimento , Pupa/genética , Temperatura , Animais , Entomologia Forense , Perfilação da Expressão Gênica
15.
Insect Biochem Mol Biol ; 139: 103652, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34562590

RESUMO

Under exposure to harmful environmental stresses, organisms exhibit a general stress response involving upregulation of the expression of heat shock proteins (HSPs) which is thought to be adaptive. Small heat shock proteins (sHSPs) are key components of this response, although shsp genes may have other essential roles in development. However, the upregulation of expression of a suite of genes under stress may not necessarily be evidence of an adaptive response to stress that involves those genes. To explore this issue, we used the CRISPR/Cas9 system to investigate pleiotropic effects of the hsp23 gene in Drosophila melanogaster. Transgenic flies carrying a pCFD5 plasmid containing sgRNAs were created to generate a complete knockout of the hsp23 gene. The transgenic line lacking hsp23 showed an increased hatch rate and no major fitness costs under an intermediate temperature used for culturing the flies. In addition, hsp23 knockout affected tolerance to hot and cold temperature extremes but in opposing directions; knockout flies had reduced tolerance to cold, but increased tolerance to heat. Despite this, hsp23 expression (in wild type flies) was increased under both hot and cold conditions. The hsp23 gene was required for heat hardening at the pupal stage, but not at the 1st-instar larval stage, even though the gene was upregulated in wild type controls at that life stage. The phenotypic effects of hsp23 were not compensated for by expression changes in other shsps. Our study shows that the fitness consequences of an hsp gene knockout depends on environmental conditions, with potential fitness benefits of gene loss even under conditions when the gene is normally upregulated.


Assuntos
Temperatura Baixa , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Regulação da Expressão Gênica , Aptidão Genética , Proteínas de Choque Térmico/genética , Temperatura Alta , Animais , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Feminino , Técnicas de Inativação de Genes , Proteínas de Choque Térmico/metabolismo , Masculino , Pupa/crescimento & desenvolvimento
16.
Sci Rep ; 11(1): 18807, 2021 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-34552169

RESUMO

Animal development relies on a sequence of specific stages that allow the formation of adult structures with a determined size. In general, juvenile stages are dedicated mainly to growth, whereas last stages are devoted predominantly to the maturation of adult structures. In holometabolous insects, metamorphosis marks the end of the growth period as the animals stops feeding and initiate the final differentiation of the tissues. This transition is controlled by the steroid hormone ecdysone produced in the prothoracic gland. In Drosophila melanogaster different signals have been shown to regulate the production of ecdysone, such as PTTH/Torso, TGFß and Egfr signaling. However, to which extent the roles of these signals are conserved remains unknown. Here, we study the role of Egfr signaling in post-embryonic development of the basal holometabolous beetle Tribolium castaneum. We show that Tc-Egfr and Tc-pointed are required to induced a proper larval-pupal transition through the control of the expression of ecdysone biosynthetic genes. Furthermore, we identified an additional Tc-Egfr ligand in the Tribolium genome, the neuregulin-like protein Tc-Vein (Tc-Vn), which contributes to induce larval-pupal transition together with Tc-Spitz (Tc-Spi). Interestingly, we found that in addition to the redundant role in the control of pupa formation, each ligand possesses different functions in organ morphogenesis. Whereas Tc-Spi acts as the main ligand in urogomphi and gin traps, Tc-Vn is required in wings and elytra. Altogether, our findings show that in Tribolium, post-embryonic Tc-Egfr signaling activation depends on the presence of two ligands and that its role in metamorphic transition is conserved in holometabolous insects.


Assuntos
Receptores ErbB/fisiologia , Proteínas de Insetos/fisiologia , Metamorfose Biológica/fisiologia , Tribolium/crescimento & desenvolvimento , Animais , Ecdisona/fisiologia , Receptores ErbB/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/genética , Larva/crescimento & desenvolvimento , Metamorfose Biológica/genética , Filogenia , Pupa/crescimento & desenvolvimento , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais , Tribolium/genética
17.
PLoS Genet ; 17(9): e1009801, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34550980

RESUMO

A key question in the rising field of neuroepigenetics is how behavioral plasticity is established and maintained in the developing CNS of multicellular organisms. Behavior is controlled through systemic changes in hormonal signaling, cell-specific regulation of gene expression, and changes in neuronal connections in the nervous system, however the link between these pathways is unclear. In the ant Camponotus floridanus, the epigenetic corepressor CoREST is a central player in experimentally-induced reprogramming of caste-specific behavior, from soldier (Major worker) to forager (Minor worker). Here, we show this pathway is engaged naturally on a large genomic scale during late pupal development targeting multiple genes differentially expressed between castes, and central to this mechanism is the protein tramtrack (ttk), a DNA binding partner of CoREST. Caste-specific differences in DNA binding of ttk co-binding with CoREST correlate with caste-biased gene expression both in the late pupal stage and immediately after eclosion. However, we find a unique set of exclusive Minor-bound genes that show ttk pre-binding in the late pupal stage preceding CoREST binding, followed by caste-specific gene repression on the first day of eclosion. In addition, we show that ttk binding correlates with neurogenic Notch signaling, and that specific ttk binding between castes is enriched for regulatory sites associated with hormonal function. Overall our findings elucidate a pathway of transcription factor binding leading to a repressive epigenetic axis that lies at the crux of development and hormonal signaling to define worker caste identity in C. floridanus.


Assuntos
Formigas/fisiologia , Pupa/crescimento & desenvolvimento , Animais , Formigas/genética , Formigas/crescimento & desenvolvimento , Epigênese Genética , Receptores Notch/metabolismo , Transcriptoma
18.
Elife ; 102021 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-34590579

RESUMO

Production of proliferative follicle cells (FCs) and quiescent escort cells (ECs) by follicle stem cells (FSCs) in adult Drosophila ovaries is regulated by niche signals from anterior (cap cells, ECs) and posterior (polar FCs) sources. Here we show that ECs, FSCs, and FCs develop from common pupal precursors, with different fates acquired by progressive separation of cells along the AP axis and a graded decline in anterior cell proliferation. ECs, FSCs, and most FCs derive from intermingled cell (IC) precursors interspersed with germline cells. Precursors also accumulate posterior to ICs before engulfing a naked germline cyst projected out of the germarium to form the first egg chamber and posterior polar FC signaling center. Thus, stem and niche cells develop in appropriate numbers and spatial organization through regulated proliferative expansion together with progressive establishment of spatial signaling cues that guide adult cell behavior, rather than through rigid early specification events.


Assuntos
Drosophila melanogaster/crescimento & desenvolvimento , Ovário/crescimento & desenvolvimento , Células-Tronco/metabolismo , Animais , Feminino , Pupa/crescimento & desenvolvimento
19.
J Invertebr Pathol ; 184: 107655, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34411606

RESUMO

The pupal soil cell of the pecan weevil, Curculio caryae (Coleoptera: Curculionidae), was reported previously to exhibit antibiosis to an entomopathogenic fungus, Beauveria bassiana. The objectives of this study were to examine 1) if the antimicrobial effect occurs in other insects that form pupal cells, 2) whether the effect extends to plant pathogenic fungi, and 3) identify the source of antibiosis in pupal soil cells of C. caryae. Antibiosis of pupal cells against B. bassiana was confirmed in-vitro in three additional curculionids, Diaprepes abbreviatus, Conotrachelus nenuphar, and Pissodes nemorensis, all of which had fewer fungal colonies relative to controls. Pupal soil cells were found to suppress phytopathogenic fungi in-vitro, including suppression of Alternaria solani by D. abbreviatus pupal cell, and that of Monilinia fructicola by C. caryae. The detection of antibiosis of soil cells formed by surface-sterilized insects using sterile soil implies the antimicrobial effect stemmed from inside the insect. Further, a novel biotic mechanism was identified: a bacterium related to Serratia nematodiphila was isolated from C. caryae pupal soil cells and was found to be associated with antibiosis. The bacterial cultures with or without autoclave had similar effects but were not as potent as pupal soil cells for suppressing B. bassiana. Also, autoclaved soil cells and autoclaved bacterial culture suppressed M. fructicola but were not as inhibitory as non-autoclaved soil cells. This indicates that antibiosis may be due to bacterial metabolites, although other factors may also be involved. Our findings suggest potential to develop the antibiotic compounds as novel bio-fungicides to control plant diseases.


Assuntos
Antibiose , Beauveria/efeitos dos fármacos , Doenças das Plantas/prevenção & controle , Serratia/fisiologia , Microbiologia do Solo , Gorgulhos/microbiologia , Animais , Fungicidas Industriais/química , Pupa/crescimento & desenvolvimento , Pupa/microbiologia , Serratia/química , Especificidade da Espécie , Gorgulhos/crescimento & desenvolvimento
20.
J Med Entomol ; 58(6): 2138-2145, 2021 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-34343277

RESUMO

Adult Calliphoridae flies, as well as their immature stages collected from carcasses, have been used as evidence in forensic investigations to estimate the postmortem interval (PMI), particularly those of the genus Chrysomya as it is one of the first genera to colonize a corpse. Chrysomya rufifacies (Macquart 1842), due to its appearance in cadaveric remains, plays a fundamental role in the study of forensic entomology. For this reason, we determined the biological cycle of C. rufifacies under semicontrolled laboratory conditions: uncontrolled average fluctuating temperature of 29. 76 ± 3. 22°C, uncontrolled average fluctuating humidity of 48. 91 ± 11.13%, and a controlled photoperiod of 12/12 (L/O). We established that the total development time from oviposition to adult emergence of C. rufifacies was 6. 5 d. The eggs took 12 h to hatch after oviposition. The complete larval stage took 60 h (instar 1 = 12 h, instar 2 = 12 h, instar 3 = 24 h, instar 3 post-feeding = 12 h). The pupa had a duration of 84 h. The species needed a total of 4642.8(±4.59) accumulated degree-hours (ADH) to complete its biological cycle.


Assuntos
Calliphoridae/crescimento & desenvolvimento , Entomologia Forense , Animais , Larva/crescimento & desenvolvimento , Óvulo/crescimento & desenvolvimento , Pupa/crescimento & desenvolvimento
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