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1.
Molecules ; 26(10)2021 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-34065875

RESUMO

The Annonaceae fruits weevil (Optatus palmaris) causes high losses to the soursop production in Mexico. Damage occurs when larvae and adults feed on the fruits; however, there is limited research about control strategies against this pest. However, pheromones provide a high potential management scheme for this curculio. Thus, this research characterized the behavior and volatile production of O. palmaris in response to their feeding habits. Olfactometry assays established preference by weevils to volatiles produced by feeding males and soursop. The behavior observed suggests the presence of an aggregation pheromone and a kairomone. Subsequently, insect volatiles sampled by solid-phase microextraction and dynamic headspace detected a unique compound on feeding males increased especially when feeding. Feeding-starvation experiments showed an averaged fifteen-fold increase in the concentration of a monoterpenoid on males feeding on soursop, and a decrease of the release of this compound males stop feeding. GC-MS analysis of volatiles identified this compound as α-terpineol. Further olfactometry assays using α-terpineol and soursop, demonstrated that this combination is double attractive to Annonaceae weevils than only soursop volatiles. The results showed a complementation effect between α-terpineol and soursop volatiles. Thus, α-terpineol is the aggregation pheromone of O. palmaris, and its concentration is enhanced by host-plant volatiles.


Assuntos
Besouros/metabolismo , Monoterpenos Cicloexânicos/análise , Monoterpenos Cicloexânicos/metabolismo , Feromônios/análise , Feromônios/metabolismo , Compostos Orgânicos Voláteis/análise , Compostos Orgânicos Voláteis/metabolismo , Animais , Annona/metabolismo , Annonaceae/metabolismo , Monoterpenos Cicloexânicos/química , Comportamento Alimentar , Cromatografia Gasosa-Espectrometria de Massas , Comportamento de Busca por Hospedeiro , Larva/metabolismo , Masculino , México , Monoterpenos/metabolismo , Olfatometria , Feromônios/química , Transdução de Sinais , Microextração em Fase Sólida , Inanição/metabolismo , Compostos Orgânicos Voláteis/química
2.
Nat Commun ; 12(1): 3362, 2021 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-34099692

RESUMO

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


Assuntos
Carbono/metabolismo , Diferenciação Celular/efeitos dos fármacos , Receptor 1 de Folato/metabolismo , Células Secretoras de Insulina/metabolismo , Leucovorina/farmacologia , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados , Animais Recém-Nascidos , Carnitina/metabolismo , Diferenciação Celular/genética , Células Cultivadas , Receptor 1 de Folato/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células Secretoras de Insulina/citologia , Larva/genética , Larva/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Camundongos , Pirimidinas/metabolismo , Suínos , Peixe-Zebra/genética
3.
Nat Commun ; 12(1): 3328, 2021 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-34099654

RESUMO

Innate behaviors consist of a succession of genetically-hardwired motor and physiological subprograms that can be coupled to drastic morphogenetic changes. How these integrative responses are orchestrated is not completely understood. Here, we provide insight into these mechanisms by studying pupariation, a multi-step innate behavior of Drosophila larvae that is critical for survival during metamorphosis. We find that the steroid-hormone ecdysone triggers parallel pupariation neuromotor and morphogenetic subprograms, which include the induction of the relaxin-peptide hormone, Dilp8, in the epidermis. Dilp8 acts on six Lgr3-positive thoracic interneurons to couple both subprograms in time and to instruct neuromotor subprogram switching during behavior. Our work reveals that interorgan feedback gates progression between subunits of an innate behavior and points to an ancestral neuromodulatory function of relaxin signaling.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Ecdisona/farmacologia , Epiderme/metabolismo , Morfogênese/efeitos dos fármacos , Neurônios/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animais , Drosophila/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Ecdisona/genética , Células Epidérmicas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular , Larva/metabolismo , Metamorfose Biológica , Morfogênese/genética , Receptores Acoplados a Proteínas G/genética , Relaxina/metabolismo
4.
Nat Commun ; 12(1): 3231, 2021 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-34050142

RESUMO

The fundamental molecular determinants by which ATP-dependent chromatin remodelers organize nucleosomes across eukaryotic genomes remain largely elusive. Here, chromatin reconstitutions on physiological, whole-genome templates reveal how remodelers read and translate genomic information into nucleosome positions. Using the yeast genome and the multi-subunit INO80 remodeler as a paradigm, we identify DNA shape/mechanics encoded signature motifs as sufficient for nucleosome positioning and distinct from known DNA sequence preferences of histones. INO80 processes such information through an allosteric interplay between its core- and Arp8-modules that probes mechanical properties of nucleosomal and linker DNA. At promoters, INO80 integrates this readout of DNA shape/mechanics with a readout of co-evolved sequence motifs via interaction with general regulatory factors bound to these motifs. Our findings establish a molecular mechanism for robust and yet adjustable +1 nucleosome positioning and, more generally, remodelers as information processing hubs that enable active organization and allosteric regulation of the first level of chromatin.


Assuntos
Montagem e Desmontagem da Cromatina , Regulação da Expressão Gênica , Histonas/metabolismo , Nucleossomos/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Regulação Alostérica/genética , Animais , DNA Fúngico/química , DNA Fúngico/genética , DNA Fúngico/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Genoma Fúngico , Histonas/genética , Histonas/isolamento & purificação , Humanos , Larva/genética , Larva/metabolismo , Conformação de Ácido Nucleico , Regiões Promotoras Genéticas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/isolamento & purificação
5.
Gene ; 790: 145688, 2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-33961974

RESUMO

In contrast to mice or zebrafish, trout exhibits post-larval muscle growth through hypertrophy and formation of new myofibers (hyperplasia). The muscle fibers are formed by the fusion of mononucleated cells (myoblasts) regulated by several muscle-specific proteins such as Myomaker or Myomixer. In this work, we identified a unique gene encoding a Myomixer protein of 77 amino acids (aa) in the trout genome. Sequence analysis and phylogenetic tree showed moderate conservation of the overall protein sequence across teleost fish (61% of aa identity between trout and zebrafish Myomixer sequences). Nevertheless, the functionally essential motif, AxLyCxL is perfectly conserved in all studied sequences of vertebrates. Using in situ hybridization, we observed that myomixer was highly expressed in the embryonic myotome, particularly in the hyperplasic area. Moreover, myomixer remained readily expressed in white muscle of juvenile (1 and 20 g) although its expression decreased in mature fish. We also showed that myomixer is up-regulated during muscle regeneration and in vitro myoblasts differentiation. Together, these data indicate that myomixer expression is consistently associated with the formation of new myofibers during somitogenesis, post-larval growth and muscle regeneration in trout.


Assuntos
Hiperplasia/patologia , Larva/citologia , Desenvolvimento Muscular , Proteínas Musculares/metabolismo , Mioblastos/citologia , Oncorhynchus mykiss/embriologia , Regeneração , Sequência de Aminoácidos , Animais , Hiperplasia/metabolismo , Larva/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas Musculares/genética , Músculo Esquelético , Mioblastos/metabolismo , Oncorhynchus mykiss/metabolismo , Filogenia , Homologia de Sequência
6.
Nat Commun ; 12(1): 3232, 2021 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-34050140

RESUMO

Arrays of regularly spaced nucleosomes dominate chromatin and are often phased by alignment to reference sites like active promoters. How the distances between nucleosomes (spacing), and between phasing sites and nucleosomes are determined remains unclear, and specifically, how ATP-dependent chromatin remodelers impact these features. Here, we used genome-wide reconstitution to probe how Saccharomyces cerevisiae ATP-dependent remodelers generate phased arrays of regularly spaced nucleosomes. We find that remodelers bear a functional element named the 'ruler' that determines spacing and phasing in a remodeler-specific way. We use structure-based mutagenesis to identify and tune the ruler element residing in the Nhp10 and Arp8 modules of the INO80 remodeler complex. Generally, we propose that a remodeler ruler regulates nucleosome sliding direction bias in response to (epi)genetic information. This finally conceptualizes how remodeler-mediated nucleosome dynamics determine stable steady-state nucleosome positioning relative to other nucleosomes, DNA bound factors, DNA ends and DNA sequence elements.


Assuntos
Montagem e Desmontagem da Cromatina , Nucleossomos/metabolismo , Sequências Reguladoras de Ácido Nucleico/genética , Animais , Proteínas de Drosophila/genética , Proteínas de Drosophila/isolamento & purificação , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Epigênese Genética , Genoma Fúngico/genética , Proteínas de Grupo de Alta Mobilidade/genética , Proteínas de Grupo de Alta Mobilidade/isolamento & purificação , Proteínas de Grupo de Alta Mobilidade/metabolismo , Histonas/genética , Histonas/metabolismo , Larva/genética , Larva/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/isolamento & purificação , Proteínas dos Microfilamentos/metabolismo , Mutagênese , Nucleossomos/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/isolamento & purificação , Proteínas de Saccharomyces cerevisiae/metabolismo , Sequenciamento Completo do Genoma
7.
Food Chem ; 359: 129911, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-33951608

RESUMO

We showed that black soldier fly larvae reared on fruits and vegetables rich in provitamin A carotenoids can accumulate significant amounts of these vitamin A precursors. Using a simulated gastro-intestinal digestion model, we demonstrated that α- and ß-carotene from the larvae are as bioaccessible as from the fruits and vegetables they were reared on. We calculated that provitamin A carotenoid-rich larvae have the capacity to provide more vitamin A than fruits and vegetables rich in these molecules. Remarkably, the incorporation of usual quantities of these larvae in feed could cover the needs of several production animals for this vitamin. Thus, our findings suggest that rearing black soldier fly larvae on by-products or waste rich in provitamin A carotenoids could be a sustainable strategy to recycle a fraction of vitamin A back into the food chain and could represent a new approach to fight against vitamin A deficiency.


Assuntos
Carotenoides/metabolismo , Dípteros , Frutas , Provitaminas/metabolismo , Verduras , Vitamina A/metabolismo , Animais , Dieta , Frutas/metabolismo , Larva/metabolismo , Verduras/metabolismo , beta Caroteno/metabolismo
8.
Aquat Toxicol ; 236: 105860, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34015756

RESUMO

Resistance to chemical insecticides including pyrethroids, the main insecticide class used against mosquitoes, has re-kindled interest in the use of neonicotinoids. In this context, the present study aimed to characterize the molecular basis of neonicotinoid resistance in the mosquito Aedes aegypti. Resistance mechanisms were studied by combining transcriptomic and genomic data obtained from a laboratory strain selected at the larval stage after 30 generations of exposure to imidacloprid (Imida-R line). After thirty generations of selection, larvae of the Imida-R line showed an 8-fold increased resistance to imidacloprid and a significant cross-tolerance to the pyrethroids permethrin and deltamethrin. Cross-resistance to pyrethroids was only observed in adults when larvae were previously exposed to imidacloprid suggesting a low but inducible expression of resistance alleles at the adult stage. Resistance of the Imida-R line was associated with a slower larval development time in females. Multiple detoxification enzymes were over-transcribed in larvae in association with resistance including the P450s CYP6BB2, CYP9M9 and CYP6M11 previously associated with pyrethroid resistance. Some of them together with their redox partner NADPH P450 reductase were also affected by non-synonymous mutations associated with resistance. Combining genomic and transcriptomic data allowed identifying promoter variations associated with the up-regulation of CYP6BB2 in the resistant line. Overall, these data confirm the key role of P450s in neonicotinoid resistance in Ae. aegypti and their potential to confer cross-resistance to pyrethroids, raising concerns about the use of neonicotinoids for resistance management in this mosquito species.


Assuntos
Resistência a Inseticidas/fisiologia , Neonicotinoides/toxicidade , Nitrocompostos/toxicidade , Aedes , Animais , Feminino , Resistência a Inseticidas/efeitos dos fármacos , Resistência a Inseticidas/genética , Inseticidas/metabolismo , Larva/metabolismo , Permetrina , Piretrinas/metabolismo , Transcriptoma/efeitos dos fármacos
9.
Aquat Toxicol ; 236: 105872, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34052719

RESUMO

Aluminium is a non-essential metal and potentially toxic to organisms whose environmental concentration increases due to pollution. In our previous studies, the behavioral changes induced by aluminium were already shown on zebrafish, a model organism widely used for ecotoxicology screening. To examine in depth the knowledge about the toxicity mechanism induced by this metal, zebrafish embryos, at 6 hpf, have been exposed to 50, 100 and 200 µM of AlCl3 for 72 h. Phenotypic alterations, apoptosis and oxidative stress responses have been assessed by evaluations of antioxidant defence and changes in metabolism at the end of treatment. The mRNA expression level of c-fos, appa and appb as marker genes of neural development and function were analyzed by qPCR for the highest used concentration. The data showed that aluminium significantly affected the development of zebrafish inducing morphological alterations and cell death. The oxidative state of larvae was altered, although the formation of reactive oxygen species and the levels of metallothioneins, and the activity of some antioxidant enzymes, decreased at the maximum concentration tested. In addition, at this concentration, the expression of the evaluated genes increased. The comprehensive information obtained gives a realistic snapshot of the aluminium toxicity and provides new information on the mechanism of action of this metal.


Assuntos
Alumínio/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Antioxidantes/metabolismo , Apoptose , Dano ao DNA , Embrião não Mamífero/metabolismo , Larva/metabolismo , Oxirredução , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismo , Peixe-Zebra/metabolismo
10.
Nat Commun ; 12(1): 2503, 2021 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-33947859

RESUMO

Habitat is one of the most important factors shaping organismal morphology, but it may vary across life history stages. Ontogenetic shifts in ecology may introduce antagonistic selection that constrains adult phenotype, particularly with ecologically distinct developmental phases such as the free-living, feeding larval stage of many frogs (Lissamphibia: Anura). We test the relative influences of developmental and ecological factors on the diversification of adult skull morphology with a detailed analysis of 15 individual cranial regions across 173 anuran species, representing every extant family. Skull size, adult microhabitat, larval feeding, and ossification timing are all significant factors shaping aspects of cranial evolution in frogs, with late-ossifying elements showing the greatest disparity and fastest evolutionary rates. Size and microhabitat show the strongest effects on cranial shape, and we identify a "large size-wide skull" pattern of anuran, and possibly amphibian, evolutionary allometry. Fossorial and aquatic microhabitats occupy distinct regions of morphospace and display fast evolution and high disparity. Taxa with and without feeding larvae do not notably differ in cranial morphology. However, loss of an actively feeding larval stage is associated with higher evolutionary rates and disparity, suggesting that functional pressures experienced earlier in ontogeny significantly impact adult morphological evolution.


Assuntos
Ecossistema , Comportamento Alimentar , Larva/crescimento & desenvolvimento , Crânio/anatomia & histologia , Animais , Anuros , Evolução Biológica , Larva/anatomia & histologia , Larva/metabolismo , Osteogênese/fisiologia , Filogenia , Análise de Componente Principal , Crânio/crescimento & desenvolvimento
11.
J Insect Sci ; 21(3)2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33974082

RESUMO

Fatty acids are important compounds for insects, but the requirements for essential fatty acids may differ between insect species. Most of the fatty acids are acquired through the insect's diet; therefore, supplementing the diet with baker's yeast (Saccharomyces cerevisiae Meyen ex E.C. Hansen), which produces unsaturated fatty acids, was predicted to affect the fatty acid composition of the insect. The tested insect was the black soldier fly (BSF) (Hermetia illucens L.), that is used as a source of protein and fat in feed. Therefore, there is importance for BSF larvae (BSFL) nutritional composition, especially the unsaturated fatty acids content, which is one of the nutritional limitations for mammalian diets. The dominant fatty acids of the tested BSFL were the saturated fatty acids: lauric, myristic, and palmitic acids, as found in other BSF studies. Oleic acid (c18:1) and linoleic acid (C18:2) were the abundant unsaturated fatty acids in the BSFL. The proportion of linoleic acid was higher in the substrate with the supplemental yeast; however, this did not affect its proportion in the larvae. The higher proportion of linoleic acid may have been exploited as a source for production of saturated lauric acid. Therefore, providing unsaturated fatty acids to the substrate through supplemental baker's yeast is not the most efficient way to increase the proportion of unsaturated fatty acids in the larvae.


Assuntos
Dípteros/química , Ácidos Graxos Insaturados , Ácidos Graxos , Saccharomyces cerevisiae , Ração Animal/análise , Animais , Dieta/veterinária , Suplementos Nutricionais , Dípteros/metabolismo , Ácidos Graxos/química , Ácidos Graxos Insaturados/química , Larva/química , Larva/metabolismo
12.
J Insect Sci ; 21(2)2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33834209

RESUMO

The aim of this review is to compile up-to-date information on the superworm, Zophobas morio (F.), regarding its biology and ecology, but also its further potential for use as a nutrient source for food and feed. We illustrate certain basic characteristics of the morphology and bio-ecology of this species, which is marginally considered as a 'pest' in durable amylaceous commodities. More recent data show that Z. morio can be a valuable nutrient and antimicrobial source that could be utilized further in insect-based feed and food production. The inclusion of this species in aquafeed has provided promising results in a wide range of feeding trials, both in terms of fish development and health. Additional data illustrate its potential for use in poultry, indicating that this species provides comparable results with those of other insect species that are used in feed. Moreover, Z. morio can be a viable waste management agent. This review aims to summarize the available data and underline data gaps for future research, toward the potential of the utilization of Z. morio for human food and animal feed. Based on the data presented, Z. morio appears to be a well-promising insect-based protein source, which potential still remains to be unfold.


Assuntos
Ração Animal , Tenebrio , Agricultura , Animais , Besouros/crescimento & desenvolvimento , Besouros/metabolismo , Proteínas de Insetos/metabolismo , Larva/crescimento & desenvolvimento , Larva/metabolismo , Nutrientes/metabolismo , Tenebrio/crescimento & desenvolvimento , Tenebrio/metabolismo
13.
Arch Insect Biochem Physiol ; 107(2): e21786, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33818830

RESUMO

Cotesia kariyai (Ck) larvae implanted into the body cavity of the Mythimna separata (armyworm) larvae get melanized and encapsulated after adhesion by hemocytes called hyperspread cells (HSCs). The present study showed that HSCs could not adhere to the implanted Ck larvae in armyworm larvae after injection of Ck polydnavirus (CkPDV) + venom (V), thus melanization and encapsulation could not occur. A C-type lectin called Mys-IML of the host armyworm larvae was considered to be involved in the recognition of foreign substances which always expressed in hemocytes. The CkPDV DNA encodes a C-type lectin called Cky811 that has high amino acid homology to Mys-IML. HSCs did not adhere when CkPDV + V was mixed with the hemolymph of armyworm larvae on glass slides and incubated in vitro, but the addition of anti-Cky811 antibody enabled HSCs to adhere. The messenger RNA (mRNA) expression of Mys-IML in armyworm larvae injected with CkPDV + V became undetectable by 6 h. On the contrary, Cky811 mRNA was well expressed in the hemocytes of armyworm larvae injected with CkPDV + V from 0.5 to 6 h. Cky811 protein was also detected in the crude extracts from Ck venom gland + Ck venom reservoir, suggesting that these proteins regulate foreign substance recognition by the armyworm within 0.5 h. These results suggest that CkPDV + V suppresses mRNA expression of Mys-IML, and that Cky811 protein expressed in hemocytes regulates foreign substance recognition of Mys-IML, resulting in inhibition of the downstream reaction steps: HSCs adhesion, melanization, and encapsulation.


Assuntos
Lectinas Tipo C/imunologia , Mariposas/parasitologia , Polydnaviridae , Vespas , Animais , Anticorpos Antivirais/metabolismo , Hemócitos/imunologia , Hemócitos/metabolismo , Interações Hospedeiro-Parasita/imunologia , Imunidade , Larva/imunologia , Larva/metabolismo , Larva/parasitologia , Lectinas Tipo C/metabolismo , Mariposas/imunologia , Polydnaviridae/metabolismo , Proteínas Virais/imunologia , Proteínas Virais/metabolismo , Vespas/patogenicidade , Vespas/virologia
14.
Acta Trop ; 219: 105917, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33839085

RESUMO

In this study, the role of nitric oxide (NO) in the pathogenesis of hydatidosis and the interaction with effects of anthelmintic drugs, albendazole and praziquantel, were examined in larval infection caused by protoscolices obtained from hydatid cysts of sheep liver in Albino Balb/c mice. Animals were divided into ten groups including controls and infected groups. Larval infection was established with intraperitoneal injection of protoscolices. Eight months after infection with protoscolices, the infected animals were divided into 6 groups. The infected animals were given a selective inhibitor of inducible nitric oxide synthase (iNOS) L-N6-(1-Iminoethyl) lysine-hydrochloride (L-NIL), NO donor sodium nitroprusside (SNP), albendazole and praziquantel as anthelmintic drugs for 7 days. In addition, control groups were composed of intact group, control, anthelmintic drugs + L-NIL, and anthelmintic drugs + SNP. The liver and blood samples were taken for cytological, histological, immunohistochemical and biochemical analyses 7 days after treatments at the end of experiment. The animals injected with protoscolices showed histopathological changes including inflammation areas, infiltration and accumulation of leukocytes, dilation of sinusoids, and damage in endothelial cells and hepatocytes at light microscopy. Electron microscopy were revealed severe damage in sinusoidal endothelial cells, leukocytes especially eosinophils in sinusoid lumens and disorganization in endoplasmic reticulum and nuclear membrane. Endothelial nitric oxide synthase (eNOS) and iNOS reactions were increased in the tissue. Anthelmintic drugs decreased inflammation areas and damages; however, it did not change NOS reactions in the animals given protoscolices. L-NIL and SNP diminished both iNOS and eNOS reactions. Unlike the group administered the inhibitor, SNP treated group exhibited less inflammation areas. Combination of these substances and drugs resulted in decreased inflammation areas. eNOS and iNOS reactions decreased in the drugs and SNP administered group, while only iNOS reaction was decreased in L-NIL given infection group. In addition, the infected groups which received SNP displayed expanded sinusoids and hepatocytes with vacuoles, intriguingly. While levels of serum nitrite/nitrate elevated only in the infection group given drugs and SNP, it decreased in the L-NIL administered group. Tissue level of malondialdehyde increased in infection groups with drugs and SNP. In conclusion, the results indicated that NO plays an important role in the pathogenesis of hydatidosis.


Assuntos
Albendazol/farmacologia , Echinococcus granulosus/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Fígado/parasitologia , Lisina/farmacologia , Óxido Nítrico/metabolismo , Praziquantel/farmacologia , Animais , Interações Medicamentosas , Echinococcus granulosus/metabolismo , Echinococcus granulosus/fisiologia , Injeções , Larva/efeitos dos fármacos , Larva/metabolismo , Larva/fisiologia , Fígado/efeitos dos fármacos , Camundongos , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Ovinos
15.
Int J Mol Sci ; 22(8)2021 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-33924375

RESUMO

Zebrafish have become a popular animal model for studying various biological processes and human diseases. The metabolic pathways and players conserved among zebrafish and mammals facilitate the use of zebrafish to understand the pathological mechanisms underlying various metabolic disorders in humans. Adipocytes play an important role in metabolic homeostasis, and zebrafish adipocytes have been characterized. However, a versatile and reliable zebrafish model for long-term monitoring of adipose tissues has not been reported. In this study, we generated stable transgenic zebrafish expressing enhanced green fluorescent protein (EGFP) in adipocytes. The transgenic zebrafish harbored adipose tissues that could be detected using GFP fluorescence and the morphology of single adipocyte could be investigated in vivo. In addition, we demonstrated the applicability of this model to the long-term in vivo imaging of adipose tissue development and regulation based on nutrition. The transgenic zebrafish established in this study may serve as an excellent tool to advance the characterization of white adipose tissue in zebrafish, thereby aiding the development of therapeutic interventions to treat metabolic diseases in humans.


Assuntos
Adipócitos/citologia , Adipócitos/metabolismo , Peixe-Zebra/genética , Peixe-Zebra/metabolismo , Tecido Adiposo/metabolismo , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Geneticamente Modificados , Forma Celular , Proteínas de Fluorescência Verde/metabolismo , Larva/genética , Larva/metabolismo , Regiões Promotoras Genéticas/genética , Transgenes , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
16.
Int J Mol Sci ; 22(9)2021 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-33919382

RESUMO

The expression of trehalase in the midgut of insects plays an important role in glucose supply to the hemolymph. Energy metabolism is usually regulated by the estrogen-related receptor (ERR). A decrease in ATP levels is caused by the ERR hindering glycolysis. However, the relationship between trehalose accumulation and ERR expression is still unclear. Here, we found that silkworm ERR (BmERR) is concentrated and BmERR expression is strongly correlated with trehalase in the midgut during the last instar silkworm larval stage. We cloned the promoter of the trehalase from Bombyx mori (BmTreh) and found that the ERR bound directly to the core response elements of the promoter. Cell level interference and the overexpression of ERR can reduce or enhance BmTreh transcription and promoter activity. Overexpressed transgenic BmERR can significantly increase the expression of BmTreh in the midgut of the last instar silkworm larvae, thereby hydrolyzing trehalose into glucose and releasing it into the hemolymph. Additionally, increased hemolymph glucose content reduces silkworm pupa weight but does not affect silk protein production from the silk gland. Our results suggest a novel function for BmERR through its involvement in BmTreh regulation and expand the understanding of ERR functions in insect trehalose metabolism.


Assuntos
Bombyx/metabolismo , Regulação Enzimológica da Expressão Gênica , Glucose/metabolismo , Hemolinfa/metabolismo , Larva/metabolismo , Receptores de Estrogênio/metabolismo , Trealase/metabolismo , Animais , Bombyx/genética , Sistema Digestório/enzimologia , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva/genética , Receptores de Estrogênio/genética , Trealase/genética , Trealose/metabolismo
17.
Arch Insect Biochem Physiol ; 107(1): e21785, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33818826

RESUMO

Mutant lethal giant larvae (lgl) flies (Drosophila melanogaster) are known to develop epithelial tumors with invasive characteristics. The present study has been conducted to investigate the influence of melatonin (0.025 mM) on behavioral responses of lgl mutant flies as well as on biochemical indices (redox homeostasis, carbohydrate and lipid metabolism, transaminases, and minerals) in hemolymph, and head and intestinal tissues. Behavioral abnormalities were quantitatively observed in lgl flies but were found normalized among melatonin-treated lgl flies. Significantly decreased levels of lipid peroxidation products and antioxidants involved in redox homeostasis were observed in hemolymph and tissues of lgl flies, but had restored close to normalcy in melatonin-treated flies. Carbohydrates including glucose, trehalose, and glycogen were decreased and increased in the hemolymph and tissues of lgl and melatonin-treated lgl flies, respectively. Key enzymes of carbohydrate metabolism showed a significant increment in their levels in lgl mutants but had restored close to wild-type baseline levels in melatonin-treated flies. Variables of lipid metabolism showed significantly inverse levels in hemolymph and tissues of lgl flies, while normalization of most of these variables was observed in melatonin-treated mutants. Lipase, chitinase, transaminases, and alkaline phosphatase showed an increment in their activities and minerals exhibited decrement in lgl flies; reversal of changes was observed under melatonin treatment. The impairment of cognition, disturbance of redox homeostasis and metabolic reprogramming in lgl flies, and restoration of normalcy in all these cellular and behavioral processes indicate that melatonin could act as oncostatic and cytoprotective agents in Drosophila.


Assuntos
Drosophila melanogaster/efeitos dos fármacos , Melatonina/farmacologia , Animais , Antineoplásicos/farmacologia , Carboidratos/sangue , Cognição/efeitos dos fármacos , Crioprotetores/farmacologia , Proteínas de Drosophila/efeitos dos fármacos , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Enzimas/sangue , Homeostase/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/genética , Larva/metabolismo , Lipídeos/sangue , Mutação , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Proteínas Supressoras de Tumor/sangue , Proteínas Supressoras de Tumor/efeitos dos fármacos , Proteínas Supressoras de Tumor/genética
18.
Int J Mol Sci ; 22(7)2021 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-33807173

RESUMO

Mealworms (Tenebrio molitor larva) are an edible insect and a protein-rich food; however, research on mealworms as a substitute protein is insufficient. In this study, mealworm fermentation extract (TMP) was assessed as a replacement for soy protein (SP) in a control diet (CON) or a high-fat diet (HFD) of mice for 12 weeks. TMP substitution reduced body weight, body weight gain, body fat mass (perirenal and mesenteric), fat size, glucose intolerance, and insulin resistance compared to the HFD-SP group. TMP alleviated hepatic steatosis (lipid contents and lipid droplets) in high-fat-fed mice and down-regulated the PPARγ, CD36, and DGAT2 gene levels. Proteomic analysis showed that a HFD for 12 weeks up-regulated 20 proteins and down-regulated 17 proteins in mice fed SP. On the other hand, TMP reversed the protein profiles. TMP significantly down-regulated KHK, GLO1, ATP5H, SOD, and DDAH1 and up-regulated DLD, Mup1, CPS1, Ces3b, PDI, and HYOU1 compared to the HFD-SP group. These proteins are involved in the glucose, lipid, and amino acid metabolism, as well as in oxidative stress and endoplasmic reticulum stress. Thus, substituting SP for TMP helped improve HFD-induced obesity, steatosis, and insulin resistance in mice. These results suggest that TMP is a potential substitute for commonly used protein sources.


Assuntos
Insetos Comestíveis/metabolismo , Obesidade/dietoterapia , Tenebrio/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Fígado Gorduroso/metabolismo , Fermentação , Intolerância à Glucose/metabolismo , Resistência à Insulina/fisiologia , Larva/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Proteínas/metabolismo , Ganho de Peso/efeitos dos fármacos
19.
Nat Commun ; 12(1): 2412, 2021 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-33893303

RESUMO

The development of a widely adopted cryopreservation method remains a major challenge in Drosophila research. Here we report a robust and easily implemented cryopreservation protocol of Drosophila melanogaster embryos. We present innovations for embryo permeabilization, cryoprotectant agent loading, and rewarming. We show that the protocol is broadly applicable, successfully implemented in 25 distinct strains from different sources. We demonstrate that for most strains, >50% embryos hatch and >25% of the resulting larvae develop into adults after cryopreservation. We determine that survival can be significantly improved by outcrossing to mitigate the effect of genetic background for strains with low survival after cryopreservation. We show that flies retain normal sex ratio, fertility, and original mutation after successive cryopreservation of 5 generations and 6-month storage in liquid nitrogen. Lastly, we find that non-specialists are able to use this protocol to obtain consistent results, demonstrating potential for wide adoption.


Assuntos
Criopreservação/métodos , Drosophila melanogaster/embriologia , Embrião não Mamífero/embriologia , Reaquecimento/métodos , Vitrificação , Animais , Crioprotetores/farmacologia , Drosophila melanogaster/genética , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/ultraestrutura , Feminino , Fertilidade/genética , Larva/genética , Larva/metabolismo , Microscopia Eletrônica , Permeabilidade/efeitos dos fármacos , Temperatura , Fatores de Tempo
20.
Nat Commun ; 12(1): 2408, 2021 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-33893307

RESUMO

Lipid shuttling between neurons and glia contributes to the development, function, and stress responses of the nervous system. To understand how a neuron acquires its lipid supply from specific lipoproteins and their receptors, we perform combined genetic, transcriptome, and biochemical analyses in the developing Drosophila larval brain. Here we report, the astrocyte-derived secreted lipocalin Glial Lazarillo (GLaz), a homolog of human Apolipoprotein D (APOD), and its neuronal receptor, the brain-specific short isoforms of Drosophila lipophorin receptor 1 (LpR1-short), cooperatively mediate neuron-glia lipid shuttling and support dendrite morphogenesis. The isoform specificity of LpR1 defines its distribution, binding partners, and ability to support proper dendrite growth and synaptic connectivity. By demonstrating physical and functional interactions between GLaz/APOD and LpR1, we elucidate molecular pathways mediating lipid trafficking in the fly brain, and provide in vivo evidence indicating isoform-specific expression of lipoprotein receptors as a key mechanism for regulating cell-type specific lipid recruitment.


Assuntos
Apolipoproteínas/metabolismo , Astrócitos/metabolismo , Encéfalo/metabolismo , Proteínas de Drosophila/metabolismo , Neuroglia/metabolismo , Neurônios/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Animais , Animais Geneticamente Modificados , Apolipoproteínas/genética , Transporte Biológico , Encéfalo/citologia , Drosophila/genética , Drosophila/metabolismo , Proteínas de Drosophila/genética , Perfilação da Expressão Gênica , Humanos , Larva/genética , Larva/metabolismo , Lipocalinas/genética , Lipocalinas/metabolismo , Ligação Proteica , Receptores Citoplasmáticos e Nucleares/genética
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