Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 918
Filtrar
1.
Exp Appl Acarol ; 78(2): 149-172, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31190248

RESUMO

Monitoring acaricide resistance and understanding the underlying mechanisms are critically important in developing strategies for resistance management and tick control. Identification of single nucleotide polymorphisms in the acaricide-resistant associated gene of Rhipicephalus microplus has enabled the development of molecular markers for detection and monitoring of resistance against different types of acaricide. There are many molecular markers developed for resistance monitoring, including mutations on target genes such as sodium channel, acetylcholinesterase, carboxylesterase, ß-adrenergic octopamine receptor, octopamine-tyramine etc. Molecular genotyping through molecular markers can detect the presence of resistance-associated genes in a tick population before it reaches high frequency. This review aims to provide an update on the various molecular markers discovered to date from different regions of the world.


Assuntos
Acaricidas/farmacologia , Proteínas de Artrópodes/genética , Resistência a Medicamentos/genética , Polimorfismo de Nucleotídeo Único/genética , Rhipicephalus/genética , Animais , Proteínas de Artrópodes/metabolismo , Mutação , Rhipicephalus/efeitos dos fármacos , Controle de Ácaros e Carrapatos
2.
Artigo em Inglês | MEDLINE | ID: mdl-31075503

RESUMO

The functions of Wnt signalling in crab limb regeneration are poorly understood. Herein, we isolated and characterised the full-length cDNA of WNT4 from Portunus trituberculatus, designated PtWNT4. The 4831 bp cDNA encodes 323 amino acid polypeptide. Protein structure prediction showed that PtWNT4 has a conserved WNT domain. The PtWNT4 gene was expressed in all regenerative limb stages, and was upregulated from stage I, with highest expression in stage III, and expression then declined in stages IV and V. Immunohistochemistry revealed strong localisation at loose connective tissue during limb regeneration, but PtWNT4 protein levels decreased upon formation of muscle fibers. Injection of WNT4 dsRNA into regenerative limbs significantly decreased PtWNT4 mRNA levels from 36 h to 5 days after injection, indicating successful gene silencing. RNA interference knockdown of PtWNT4 expression greatly retarded limb regeneration compared with the control group. Blastema emergence phenotype analysis revealed limb regeneration rates of 0, 31.5% and 40.5% for the dsWNT4 group at 36 h, 3 days and 5 days, respectively, compared with 29.95%, 83.0% and 92.5% for the saline-injected control groups (p <0.05). Expression analysis on the WNT4 using RNAi provide important information for understanding its functional mechanism during limb regeneration in P. trituberculatus. The results also contribute to our understanding of the role of Wnt signalling during limb regeneration in crustaceans.


Assuntos
Proteínas de Artrópodes/metabolismo , Braquiúros/metabolismo , Membro Posterior/fisiologia , Interferência de RNA , Regeneração/fisiologia , Proteína Wnt4/metabolismo , Animais , Proteínas de Artrópodes/genética , Braquiúros/genética , Proteína Wnt4/genética
3.
Exp Appl Acarol ; 77(4): 527-543, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31062204

RESUMO

Biological control of spider mites in hot and dry weather is a serious technical issue. A high-temperature adapted strain (HTAS) of the predatory mite Neoseiulus barkeri Hughes was selected from its conventional strain (CS), via long-term heat acclimation and frequent heat hardenings in our previous studies. However, the environment of high temperature is usually associated with enhanced ultraviolet (UV) radiation. In the present study, the physiological effects of UV-B radiation on survival rate and egg damage of N. barkeri were investigated, as well as the activities and expression profiles of antioxidant enzymes to UV-B radiation stress. UV-B radiation had deleterious effects on egg hatchability and survival of N. barkeri. Adults of the HTAS strain were less UV-B resistant than those of the CS strain; they also had lower levels of enzymatic activity of superoxide dismutase (SOD) and catalase against oxidative damage and weaker upregulation of SOD genes. The mRNA expression of three SOD genes of CS adult females immediately increased whereas that of HTAS showed almost no difference under UV-B stress for 1 h. The results showed the HTAS of N. barkeri had lower fitness under UV-B stress compared with the CS of N. barkeri. These results suggested that long-term heat acclimation may exert a profound impact on the developmental physiology of N. barkeri.


Assuntos
Proteínas de Artrópodes/genética , Aptidão Genética/efeitos da radiação , Ácaros/efeitos da radiação , Comportamento Predatório/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Adaptação Biológica , Animais , Antioxidantes/metabolismo , Proteínas de Artrópodes/metabolismo , Feminino , Temperatura Alta , Longevidade/efeitos da radiação , Ácaros/enzimologia , Ácaros/genética , Ácaros/fisiologia , Óvulo/fisiologia , Óvulo/efeitos da radiação , Controle Biológico de Vetores , Transcrição Genética/efeitos da radiação
4.
Sci Total Environ ; 683: 193-201, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31129327

RESUMO

DMSO is a very common solvent for hydrophobic chemicals that may pose a threat to aquatic organisms. Ectoine (ECT) is a protective amino acid produced by various strains of halophilic bacteria with high potential to alleviate detrimental effects induced by environmental stressors. This amino acid is used in many cosmetics and pharmaceuticals may enter aquatic ecosystems interacting with ions and macromolecules. Little is known on the effects of DMSO and its interaction with ECT on behavioral, physiological and biochemical endpoints of aquatic invertebrates. Therefore, the purpose of the present study was to determine protective effects of DMSO alone and in the combination with ECT on hopping frequency, swimming speed, heart rate, thoracic limb activity, catalase activity and NOx level in an animal model, Daphnia magna subjected to 0.1% and 1% DMSO alone and during combinatorial exposure to ECT (0-25 mg/L) and DMSO for 24 h and 48 h. The results showed that swimming speed, heart rate and thoracic limb activity were inhibited by both 0.1% and 1% DMSO alone however alleviating effects were observed in the combination DMSO + ECT. Thoracic limb activity was higher in the animals exposed to both solutions of DMSO alone, however the parameter was more stimulated at DMSO + ECT. The results suggest that DMSO alone may alter Daphnia behavior and physiological parameters, therefore use of the control group of non-treated animals with DMSO alone would be recommended to avoid data misinterpretation.


Assuntos
Diamino Aminoácidos/farmacologia , Daphnia/efeitos dos fármacos , Dimetil Sulfóxido/toxicidade , Substâncias Protetoras/farmacologia , Poluentes Químicos da Água/toxicidade , Animais , Proteínas de Artrópodes/metabolismo , Catalase/metabolismo , Daphnia/enzimologia , Daphnia/fisiologia , Frequência Cardíaca/efeitos dos fármacos , Locomoção/efeitos dos fármacos , Movimento/efeitos dos fármacos , Óxido Nítrico/metabolismo , Natação
5.
Chem Biol Interact ; 307: 29-36, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30991043

RESUMO

Carbonyl reductases (CRs) represent a fundamental enzymatic defense mechanism against oxidative stress. While commonly two carbonyl reductases (CBR1 and CBR3) are found in mammalian genomes, invertebrate model organisms like Drosophila melanogaster express no CR but a functional homolog to human CBR1, termed sniffer. The importance of sniffer could be demonstrated in D. melanogaster where it protected against age-dependent neurodegeneration. Interestingly, the microcrustacean Daphnia harbors four copies of the CR gene (CR1, CR2, CR3, CR4) in addition to one sniffer gene. Due to this unique equipment Daphnia is an ideal model organism to investigate the function of sniffer. Recombinant sniffer from D. magna und D. pules were produced in E. coli, purified by Ni-affinity chromatography and tested with a variety of aliphatic and aromatic diketones, reactive aldehydes and precursors of advanced glycation end products (AGE). The highest catalytic activities were determined for sniffer from D. pulex with the aromatic dicarbonyls 9,10-phenanthrenequinone (kcat/Km = 2.6 s-1 x µM-1) and isatin (kcat/Km = 1.5 s-1 x µM-1). While sniffer from D. magna displayed preference for the same two substances, the respective catalytic activities were noticeably lower. Kinetic constants with aliphatic diketones were generally lower than those with aromatic dicarbonyls for both sniffer enzymes. The best aliphatic diketone as substrate for sniffer from D. magna and D. pulex was hexane-3,4-dione with kcat/Km = 0.23 s-1 µM-1 and kcat/Km = 0.35 s-1 µM-1, respectively. Poor or no detectable activity of the two sniffer enzymes was seen with the aliphatic diketones 2,5-hexanedione and 3,5-heptanedione, the aldehydes butanal, hexanal, decanal, crotonaldehyde, acrolein, trans-2-hexenal, and the AGE precursors glyoxal, methylglyoxal, furfural and glyceraldehyde, indicating no physiological function in the metabolism of short-chain aldehydes. Substrate inhibition for both sniffer enzymes was observed with the quinone substrates 1,4-naphthoquinone and 2-methyl-1,4-benzoquinone. From a variety of pesticides endosulfan turned out as an effective inhibitor of the sniffer enzymes (Ki = 9.2 µM for sniffer from D. magna, Ki = 12.0 µM for sniffer from D. pulex). In conclusion, the present results on sniffer from the protein superfamily of the short-chain dehydrogenases/reductases (SDR) in Daphnia ssp. complement earlier studies on carbonyl reductases in the same species and indicate that Daphnia is an interesting model to study the overall response to carbonyl stress.


Assuntos
Oxirredutases do Álcool/metabolismo , Proteínas de Artrópodes/metabolismo , Clonagem Molecular , Oxirredutases do Álcool/antagonistas & inibidores , Oxirredutases do Álcool/genética , Animais , Proteínas de Artrópodes/antagonistas & inibidores , Proteínas de Artrópodes/genética , Biocatálise , Daphnia/enzimologia , Proteínas de Drosophila/antagonistas & inibidores , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/enzimologia , Endossulfano/química , Endossulfano/metabolismo , Cinética , Fenantrenos/química , Fenantrenos/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Especificidade por Substrato
6.
Arthropod Struct Dev ; 50: 43-52, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30974153

RESUMO

Naupliar development in copepods includes the generation of usually five pairs of post-mandibular segments. Since copepod nauplii show no outer body articulation, the only indication of larval segmentation is the expression of limb buds. Yet, in copepods the timing and sequence of limb bud expression in larval development varies to a large degree. In harpacticoid nauplii for instance, the 1st maxillae are formed at an early naupliar stage. By contrast, the four remaining pairs of limb buds frequently appear simultaneously with the last naupliar stage. The complete process of larval segment formation takes place under the body surface and has never been described in detail. To broaden our knowledge of early segmentation in copepods, we here describe the segmentation of the harpacticoid nauplius Tigriopus californicus by analysing the expression of the segment marker Engrailed and uncover the sequential addition of seven post-mandibular segments. The stripe formation and arrangement of labelled cells corresponds largely to those of other crustaceans studied in this respect. Together with a morphological approach using histology, SEM, and 3D-reconstructions based on CLSM we solve the so far controversial identity of the external limb buds in the final naupliar stage. In contrast to previous studies, we can show that all limb pairs from the 1st maxillae to the 3rd thoracopods are formed. Yet, the anlage of the maxilliped (1st thoracopod) remains hidden underneath the cuticle being never externally expressed in the nauplius.


Assuntos
Padronização Corporal , Copépodes/crescimento & desenvolvimento , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Padronização Corporal/genética , Copépodes/genética , Perfilação da Expressão Gênica , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Microscopia Confocal , Microscopia Eletrônica de Varredura , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
7.
Fish Shellfish Immunol ; 89: 108-116, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30928665

RESUMO

To identify molecules involved in Macrobrachium rosenbergii nodavirus (MrNV) entry into hemocytes of the giant freshwater prawn M. rosenbergii, biotinylated prawn hemocyte membrane proteins were prepared, purified and separated by SDS-PAGE. The proteins were blotted on the nitrocellulose membrane before incubation with the MrNV capsid protein (MrNV-CP) by a VOPBA technique. Subsequent mass spectrometry and analysis of immune-reactive bands represent putative binding partners including transglutaminase (TG), actin, α2-macroglobulin, α1-tubulin, F1-ATP synthase ß-subunit and a currently uncharacterized protein. The sequence of TG has been characterized and found 5 amino acids differences to a previously reported MrTG (ADX99580), mainly at its N-terminal part and thus, we named it MrTGII (KM008611). Recombinant MrTGII was prepared to produce a polyclonal antibody against it, which was successfully revealed the presence of MrTGII (100 kDa) in prawn hemocyte lysates. Using the pentylamine-biotin incorporation assay, an acyl transfer reaction was observed when hemocyte lysates were added to solutions containing MrNV-CP, suggesting that hemocyte MrTG could use MrNV-CP as the substrate. The expression levels of MrTGII were changed during the course of MrNV infection. By using immunostaining technique, location of MrTGII on the hemocyte surface was confirmed. Specific interaction between MrTGII with MrNV-CP in a dose-dependent manner was confirmed by in vitro ELISA assay. The highest binding activity of MrNV-CP was found with the N-terminal portion of the protein. In vitro neutralization using anti-MrTGII antibody resulted in inhibition of MrNV attachment to the hemocyte surface, accompanied by a dramatic reduction in viral replication. This is the first time that crustacean TG has been shown to be involved in viral entry, in addition to its roles in blood clotting and haematopoiesis.


Assuntos
Hemócitos/enzimologia , Nodaviridae/fisiologia , Palaemonidae/imunologia , Transglutaminases/genética , Replicação Viral , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Hemócitos/virologia , Microscopia de Fluorescência , Transglutaminases/química , Transglutaminases/metabolismo
8.
Food Chem ; 290: 277-285, 2019 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-31000048

RESUMO

Fluorescein-isothiocyanate (FITC) labeled trypsin-like protease was prepared and injected into the hepatopancreas of white shrimp. Different segments of the injected shrimp were analyzed with a fluorescence microscope during storage. FITC-trypsin-like protease can be detected in the first segment of shrimp muscle at day 4, while it cannot be observed in the second segment until day 6. The results showed that trypsin-like protease can migrate from hepatopancreas to the tail portion. Texture profile analysis showed that soybean trypsin inhibitor retarded the softening of the shrimp muscle. The rheological results revealed that the content of myosin heavy chain (MHC) in shrimp muscle was decreased with the extended storage time. Proteomics analysis displayed that trypsin-like protease accelerated the metabolism of postmortem muscle. It can be concluded that trypsin-like protease migrated from the hepatopancreas to the muscle tissue, degraded myofibrillar protein, deteriorated the muscle texture, and eventually leaded to the softening of white shrimp.


Assuntos
Proteínas de Artrópodes/metabolismo , Músculos/fisiologia , Penaeidae/metabolismo , Peptídeo Hidrolases/metabolismo , Animais , Proteínas de Artrópodes/química , Eletroforese em Gel Bidimensional , Hepatopâncreas/metabolismo , Focalização Isoelétrica , Músculos/efeitos dos fármacos , Peptídeo Hidrolases/química , Mudanças Depois da Morte , Proteoma/análise , Inibidores da Tripsina/farmacologia
9.
Fish Shellfish Immunol ; 90: 363-375, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30974219

RESUMO

Superoxide dismutases (SODs) are important antioxidant enzymes that occur in virtually all oxygen-respiring organisms, and copper/zinc SOD (Cu/ZnSOD) is one of the most important SODs. In the present study, Macrobrachium rosenbergii Cu/Zn-SOD was expressed in a yeast eukaryotic system. The open reading frame (ORF) of MrCu/ZnSOD was cloned into the plasmid vector pHAC181, and the recombinant plasmid was integrated into the downstream region of the GAL1 promoter in Saccharomyces cerevisiae strain GAL1-ScRCH1 via homologous recombination. The resulting recombinant MrCu/ZnSOD consisted of a 3 × HA-tag at its C-terminal. Via western blot, the molecular weight of the recombinant MrCu/ZnSOD was estimated at about 30 kDa. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of this recombinant MrCu/ZnSOD ranged from 0.556 to 0.840 µM, and from 0.967 to 2.015 µM, respectively. The recombinant MrCu/ZnSOD protein was able to agglutinate four Gram-negative bacterial strains, as well as two of three Gram-positive strains (except Staphylococcus aureus). This demonstrated that the recombinant protein possessed some antimicrobial activity against certain Gram-positive and Gram-negative bacteria. M. rosenbergii were fed with the recombinant yeast strain MrCu/ZnSOD for 4 weeks and then challenged with the most common crustacean pathogen, Vibrio parahaemolyticus. This group of prawns presented lower mortality, higher enzymatic activity, and higher expression of the mRNA of immune-related genes than that in the control groups. Taken together, these results suggest that MrCu/ZnSOD is an antioxidant enzyme and antimicrobial peptide involved in the crustacean innate immune system and offers protection to the host against pathogenic bacteria.


Assuntos
Palaemonidae/genética , Palaemonidae/imunologia , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/imunologia , Vibrio parahaemolyticus/imunologia , Aglutinação , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/metabolismo , Bactérias Gram-Negativas/imunologia , Bactérias Gram-Positivas/imunologia , Microrganismos Geneticamente Modificados/metabolismo , Palaemonidae/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Superóxido Dismutase-1/metabolismo
10.
Int J Mol Sci ; 20(7)2019 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-30965605

RESUMO

The doublesex and mab-3 related transcription factor (DMRT) gene family involvement in sex development is widely conserved from invertebrates to humans. In this study, we identified a DM (Doublesex/Mab-3)-domain gene in Macrobrachium nipponense, which we named MniDMRT11E because it has many similarities to and phylogenetically close relationships with the arthropod DMRT11E. Amino acid alignments and structural prediction uncovered conservation and putative active sites of the DM domain. Real-time PCR analysis showed that the MniDMRT11E was highly expressed in the ovary and testis in both males and females. Cellular localization analysis showed that DMRT11E was mainly located in the oocytes of the ovary and the spermatocyte of the testis. During embryogenesis, the expression level of MniDMRT11E was higher at the cleavage stage than at other stages. During the different stages of ovarian development, MniDMRT11E expression gradually increased from OI to OIII and decreased to the lowest level at the end of OIV. The results indicated that MniDMRT11E probably played important roles in embryonic development and sex maturity in M. nipponense. MniDMRT11E dsRNA injection also significantly reduced vitellogenin (VG) expression and significantly increased insulin-like androgenic gland factor (IAG) expression, indicating a close relationship in gonad development.


Assuntos
Proteínas de Artrópodes/metabolismo , Palaemonidae/embriologia , Palaemonidae/metabolismo , Animais , Proteínas de Artrópodes/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Masculino , Ovário/embriologia , Ovário/metabolismo , Palaemonidae/genética , Testículo/embriologia , Testículo/metabolismo
11.
Fish Shellfish Immunol ; 89: 603-613, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30902724

RESUMO

In order to study the effects of Moringa oleifera leaf extract on Macrobrachium rosenbergii under high ammonia exposure, freshwater prawns were randomly divided into five groups: a control group was fed with basal diet, and four treatment groups fed with basal diet supplemented with 0.25%, 0.5% and 1.0% M. oleifera leaf extract and 0.025% Enrofloxacin for 60 days, respectively. Then, freshwater prawns were exposed to high ammonia stress for 72 h and Vibro anguillarum infection. The growth, antioxidant capabilities, related immune genes as well as resistance to infection by V. anguillarum were determined. The results showed that compared with the control group, the weight gain, specific growth rate and protein efficiency rate, haemolymph catalase (CAT), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) increased while feed conversion ratio, haemolymph aspartate aminotransferase, alanine aminotransferase, nitrogen oxide (NO), hepatopancreas heat shock proteins (HSP70), immune deficiency (IMD) expression levels decreased in the group of 0.5% M. oleifera leaf extract before the stress. After ammonia stress, the group of 0.5% M. oleifera leaf extract also could improve the haemolymph SOD, glutathione peroxidase, NO, iNOS, hepatopancreas HSP70 expression levels and reduce haemolymph CAT, hepatopancreas peroxiredoxin 5 and NF kappa B inhibitor alpha expression level compared with the control group. The rate of mortality of the prawns challenged with V. anguillarum was lower in the supplemented groups in comparison with the control group with the lowest being in the group of 0.5% M. oleifera leaf extract. Antioxidant activities as well as biochemical parameters in the enrofloxacin group (0.025%E) were not significantly enhanced both pre and post challenge in comparison with the M. oleifera leaf extract groups, showing the superiority of the natural herb over the synthetic antibiotic. In summary, this study suggested that at an inclusion rate of 0.5%, M. oleifera leaf extract could increase the growth performance, even has positive effects on physiological and immune function and prevents high ammonia stress in the Freshwater prawn, M.rosenbergii.


Assuntos
Amônia/efeitos adversos , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/imunologia , Moringa oleifera/química , Palaemonidae/efeitos dos fármacos , Extratos Vegetais/metabolismo , Vibrio/efeitos dos fármacos , Ração Animal/análise , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Dieta , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Palaemonidae/genética , Palaemonidae/crescimento & desenvolvimento , Palaemonidae/imunologia , Extratos Vegetais/administração & dosagem , Folhas de Planta/química , Distribuição Aleatória , Estresse Fisiológico/efeitos dos fármacos , Vibrio/fisiologia
12.
Int Immunopharmacol ; 70: 216-224, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30851701

RESUMO

OBJECTIVES: House dust mites, including Der p1, are common allergens. The current study was designed to explore the allergen-specific immune tolerance effects of Der p1-modified dendritic cells (DCs) through IL-4, IL-10 and IL-13 on an allergic rhinitis (AR) mouse model. METHODS: A lentivirus was modified to express Derp1. Then, immature DCs from mice were infected with this modified lentivirus to generate a lenti-Derp1-GFP DCs. 24 mice were random divided into four groups (n = 6 each), AR mouse were sensitized by Derp1 allergens and treated with lenti-GFP DCs (GFP-DC/AR group), or lenti-Derp1-GFP DCs (Der p1-DC/AR group) and dexamethasone (Dex/AR group), mice in the control group were treated with PBS instead of Der p1 then also intraperitoneally injected with 5 × 106 lenti-GFP DCs/mouse. AR symptoms expressed by each mouse were recorded. The proportions of CD4+CD25+Foxp3+ regulatory T cells among CD4+ T cells in the peripheral blood, and mRNA and protein expression levels of IL-4, IL-10, and IL-13 were measured. RESULTS: DCs infected with lenti-Derp1-GFP stimulated the maturation of DCs. Compared with the GFP-DC/AR group, mice in the Der p1-DC/AR group showed an ameliorated allergic response, a significant decrease in the levels of serum IgE, IgG1, and histamine, and a decrease in the expression of IL-4 and IL-13 mRNA and protein in the nasal mucosa. The expression of IL-10 increased in the Der p1-DC/AR group to a level similar to that observed in the Dex/AR group. CONCLUSIONS: These results indicate that Der p1-modified DCs have therapeutic potential for AR via downregulation of IL-4 and IL-13, and upregulation of IL-10.


Assuntos
Alérgenos/metabolismo , Antígenos de Dermatophagoides/metabolismo , Proteínas de Artrópodes/metabolismo , Cisteína Endopeptidases/metabolismo , Células Dendríticas/fisiologia , Mucosa Nasal/fisiologia , Rinite Alérgica/imunologia , Alérgenos/genética , Animais , Antígenos de Dermatophagoides/genética , Proteínas de Artrópodes/genética , Células Cultivadas , Cisteína Endopeptidases/genética , Modelos Animais de Doenças , Vetores Genéticos , Humanos , Tolerância Imunológica/genética , Interleucina-10/metabolismo , Interleucina-13/metabolismo , Interleucina-4/metabolismo , Lentivirus/genética , Camundongos , Camundongos Endogâmicos BALB C , Pyroglyphidae/imunologia
13.
Fish Shellfish Immunol ; 88: 1-8, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30826412

RESUMO

In this study, GST Pull-down and mass spectrometry was applied to the precipitation and identification of the small GTP-binding protein (Rab7) interacting protein in hemocyte of Litopenaeus vannamei. According to the search in GenBank with the peptide mass fingerprint, the 45 kDa protein which was pulled down with the GST-tagged Rab7 (GST-Rab7, GTP bound form) was identified to be ß-actin with 28% coverage of amino acid sequences. The interaction of Rab7 with ß-actin was verified by both GST Pull-down and ELISA in vitro. Meanwhile, confocal microscopic observation showed that Rab7 could be co-localized with ß-actin in hemocytes at 12 h post white spot syndrome virus (WSSV) infection (hpi). GST Pull-down and western blotting were used to analyze the cross-interaction between WSSV VP28, Rab7 and ß-actin. The results showed that the GST-VP28, His-tagged Rab7 (His-Rab7) and His-ß-actin formed a tripartite complex. At 12 hpi, confocal microscopic observation showed that WSSV could be co-localized with Rab7 and ß-actin in hemocytes respectively. Furthermore, based on the in vivo neutralization assay, recombinant His-ß-actin accelerated the infection of WSSV, conversely, recombinant His-Rab7 delayed WSSV infection in shrimp. These results suggested the interaction of Rab7 with ß-actin and this interaction was involved in WSSV infection.


Assuntos
Penaeidae/virologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Proteínas rab de Ligação ao GTP/metabolismo , Actinas , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/metabolismo , Hemócitos/virologia , Microscopia Confocal , Penaeidae/metabolismo
14.
Dev Comp Immunol ; 95: 38-49, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30738078

RESUMO

The p38 mitogen-activated protein kinases (MAPKs) are evolutionally conserved from yeasts to mammals, and are involved in the regulation of cells response to various extracellular stimuli. In this study, the p38 MAPK gene (designated as Spp38) of mud crab (Scylla paramamosain) was identified and studied. Spp38 contained the conserved Thr-Gly-Tyr (TGY) motif and a Ala-Thr-Arg-Trp (ATRW) substrate-binding site. Spp38 transcript was ubiquitously expressed in all tissues examined, with the highest expression found in muscle and hepatopancras. Quantitative real-time PCR revealed that Spp38 was upregulated in hemocytes and hepatopancras after infection with Vibrio parahemolyticus and Lipopolysaccharides (LPS). Reporter gene assays indicated that Spp38 activated the expression of anti-lipopolysaccharides (SpALF1 - SpALF6) in S. paramamosian. RNA interference (RNAi)-mediated knockdown of Spp38 or inhibition of Spp38 by SB203580 decreased the expression levels of SpALF1-6 and dual oxidase (SpDuox1 and SpDuox2) in S. paramamosian, which consequently reduced reactive oxygen species (ROS) production thereby significantly increasing the bacterial count in the hemolymph of mud crabs. Similarly, there was a significant reduction in bacterial clearance ability of hemolymph after Spp38 knockdown followed by V. parahemolyticus infection. Taken together, the current data indicated that Spp38 could play a vital role in maintaining the homeostasis of hemolymph microbiota in S. paramamosain.


Assuntos
Proteínas de Artrópodes/imunologia , Braquiúros/imunologia , Hemolinfa/microbiologia , Microbiota/imunologia , Proteínas Quinases p38 Ativadas por Mitógeno/imunologia , Animais , Proteínas de Artrópodes/metabolismo , Braquiúros/microbiologia , Hemócitos/imunologia , Hemócitos/metabolismo , Hemolinfa/citologia , Hemolinfa/imunologia , Hepatopâncreas/metabolismo , Homeostase , Lipopolissacarídeos/imunologia , Músculos/metabolismo , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/imunologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
15.
Cell Stress Chaperones ; 24(2): 385-392, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30701477

RESUMO

The crustacean, Artemia franciscana, displays a complex life history in which embryos either arrest development and undertake diapause as cysts or they develop into swimming nauplii. Diapause entry is preceded during embryogenesis by the synthesis of specific molecular chaperones, namely the small heat shock proteins p26, ArHsp21, and ArHsp22, and the ferritin homolog, artemin. Maximal synthesis of diapause-specific molecular chaperones is dependent on the transcription factor, heat shock factor 1 (Hsf1), found in similar amounts in cysts and nauplii newly released from females. This investigation was performed to determine why, if cysts and nauplii contain comparable amounts of Hsf1, only cyst-destined embryos synthesize diapause-specific molecular chaperones. Quantification by qPCR and immunoprobing of Western blots, respectively, demonstrated that hsf1 mRNA and Hsf1 peaked by day 2 post-fertilization in embryos that were developing into cysts and then declined. hsf1 mRNA and Hsf1 were present in nauplii-destined embryos on day 2 post-fertilization, but in much smaller amounts than in cyst-destined embryos, and they increased in quantity until release of nauplii from females. Immunofluorescent staining revealed that the amount of Hsf1 in nuclei was greatest on day 4 post-fertilization in cyst-destined embryos but could not be detected in nuclei of nauplius-destined embryos at this time. The differences in quantity and location of Hsf1 explain why embryos fated to become cysts and eventually enter diapause synthesize p26, ArHsp21, ArHsp22, and artemin, whereas nauplius-destined embryos do not produce these molecular chaperones.


Assuntos
Artemia/embriologia , Proteínas de Artrópodes/metabolismo , Diapausa , Embrião não Mamífero/metabolismo , Fatores de Transcrição de Choque Térmico/metabolismo , Animais , Artemia/metabolismo , Proteínas de Artrópodes/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Transcrição de Choque Térmico/genética , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Oocistos/crescimento & desenvolvimento , Oocistos/metabolismo
16.
Fish Shellfish Immunol ; 87: 144-154, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30630047

RESUMO

The oriental river prawn Macrobrachium nipponense is a highly adaptable, tolerant, and fecund freshwater prawn that inhabits a wide range of aquatic environments. The hepatopancreas of crustaceans is not only a site for secretion of digestive enzymes, and also plays important roles in several metabolic processes, such as lipid and carbohydrate metabolism. It is the main organ for the detoxification and immunity. In this study, high-throughput sequencing techniques were used to detect the effect of nitrite stress (10 mg/L nitrite-N for 48 h) on gene expression in the hepatopancreas of M. nipponense. A total of 13,769 million reads were harvested, and 94,534 transcripts were de novo assembled using Trinity software and produced 56,054 non-redundant transcripts. A total of 825 differentially expressed genes were obtained comparing 48 h nitrite stress with control group. In the analysis of GO and KEGG database, significant differences were found in 49 pathways. Immune-related pathways under nitrite stress included arginine and proline metabolism, glutamate metabolism, Jak-Stat signaling pathway, endocytosis, wnt signaling pathway, RIG-I-like receptor signaling pathway, TGF-beta signaling pathway, GnRH signaling pathway and phagosome. Apoptosis-related pathway was also significantly altered, such as lysosome and apoptosis. Remarkably, nitrite stress altered the expression patterns of key apoptosis genes (tetraspanins-like protein, LAMP, CD63, caspase 3C and Caspase 1) and immune genes (Serine proteinase-like protein, C-type lectin, daf-36, SOCS-2, alpha-2-macroglobulin), confirmed that nitrite-stress induce immune response and eventually even apoptosis. This study provided a new insight into the role of hepatopancreas in crustaceans, and further investigation will continue.


Assuntos
Hepatopâncreas/efeitos dos fármacos , Nitritos/toxicidade , Palaemonidae/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Apoptose/efeitos dos fármacos , Proteínas de Artrópodes/metabolismo , Perfilação da Expressão Gênica , Hepatopâncreas/metabolismo , Imunidade Inata/efeitos dos fármacos , Palaemonidae/genética , Palaemonidae/metabolismo , Transdução de Sinais , Estresse Fisiológico
17.
Dev Comp Immunol ; 94: 16-21, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30641118

RESUMO

C1q is an important immune gene that can mediate a variety of immune regulatory functions, and is involved in complement pathway activation. In the present study, a ghC1q gene from the razor clam Sinonovacula constricta was identified and named ScghC1q-1. The complete ScghC1q-1 gene is 692 bp in length, with an open reading frame (ORF) of 489 bp encoding a protein of 162 amino acids. ScghC1q-1 mRNA was widely expressed in various tissues, and transcript levels in the hemolymph were significantly up-regulated following Staphylococcus aureus or Vibrio anguillarum challenge. Recombinant ScghC1q-1 protein was found to agglutinate both Gram-positive and Gram-negative bacteria. These results indicate that ScghC1q-1 plays an essential role in the immune defense of S. constricta.


Assuntos
Proteínas de Artrópodes/genética , Bivalves/imunologia , Complemento C1q/genética , Hemolinfa/fisiologia , Infecções Estafilocócicas/imunologia , Staphylococcus aureus/fisiologia , Vibrioses/imunologia , Vibrio/fisiologia , Animais , Proteínas de Artrópodes/metabolismo , Clonagem Molecular , Complemento C1q/metabolismo , Perfilação da Expressão Gênica , Imunidade Inata , Filogenia , Alinhamento de Sequência , Regulação para Cima
18.
Dev Comp Immunol ; 95: 68-76, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30682447

RESUMO

Tyramine (TA), a neuroactive chemical, plays various important physiological roles in insects by activating distinct G-protein-coupled receptors (GPCRs). In this study, we investigated the effects of by pharmacological injection of TA on immune resistance regulation and its signal pathway in white shrimp Litopenaeus vannamei. Results showed significant increases in the total haemocyte count (THC), semigranular cells (SGCs), granular cells (GCs), phenoloxidase (PO) activity per 50 µL of haemolymph and respiratory bursts (RBs) at 0.5, 1, 2 and/or 4 h; hyaline cells (HCs) at 0.5 h, as well as phagocytic activity (PA) and clearance efficiency (CE) at 2, 4 and/or 8 h, but significantly decreased PO activity per granulocyte at 0.5-2 h for shrimp injected with TA at 100 and 1000 pmol shrimp-1. Plasma lysozyme activities of TA-injected shrimp were significantly higher than those of the saline control at 1 h. All of the immune parameters had returned to control levels by 8 h after receiving TA except the clearance efficiency, which had returned to its control value by 16 h. The TA injection also significantly decreased the mortality of shrimp challenged with Vibrio alginolyticus. Furthermore, immune parameters of shrimp that received TA at 1000 pmol shrimp-1 for 1 h were higher than those of shrimp that received the saline. The upregulating effect of TA was blocked by co-injection with phentolamine (Phe) in terms of the THC, HC, SGCs, PO activity, PA and CE; by co-injection with prazosin (Pra) in terms of the THC, HC, SGCs, PO activity, PA and CE; by co-injection with propranolol (Prop) in terms of the PA and CE; and by co-injection with metoprolol (Meto) in terms of the THC and SGCs. The most potent effect in immunocompetence of tested antagonists was Pra, and except for circulating haemocyte, it was Phe. These results suggest that ≤1000 pmol shrimp-1 of a TA injection mediates transient upregulation of immunity, which in turn promotes the resistance of L. vannamei to V. alginolyticus, and the active effects are mediated via octopamine/tyramine (OA/TA) receptors.


Assuntos
Proteínas de Artrópodes/metabolismo , Resistência à Doença/imunologia , Penaeidae/imunologia , Transdução de Sinais/efeitos dos fármacos , Tiramina/farmacologia , Animais , Aquicultura , Proteínas de Artrópodes/imunologia , Resistência à Doença/efeitos dos fármacos , Hemócitos/efeitos dos fármacos , Hemócitos/imunologia , Hemócitos/metabolismo , Imunidade Inata/efeitos dos fármacos , Penaeidae/efeitos dos fármacos , Penaeidae/metabolismo , Penaeidae/microbiologia , Receptores de Amina Biogênica/imunologia , Receptores de Amina Biogênica/metabolismo , Transdução de Sinais/imunologia , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrio alginolyticus/imunologia , Vibrio alginolyticus/patogenicidade
19.
Physiol Biochem Zool ; 92(2): 152-162, 2019 Mar/Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30694107

RESUMO

Thermal stress experiments are essential for understanding organisms' thermal limits and the physiological processes that contribute to establishing those limits. Experiments typically employ either an abrupt transfer to near-lethal temperatures or a gradually increasing thermal exposure. In the current study, we used three populations of the intertidal copepod Tigriopus californicus that are known to differ in upper thermal tolerance to investigate the effects of gradual versus abrupt thermal exposures on survivorship, developmental time, and heat shock protein gene expression. The developmental rate of nauplii was unaffected following the gradual exposure, whereas developmental time slowed by ∼2 d (∼20%) following an abrupt exposure. The gradual exposure also improved survivorship in comparison to the abrupt exposure. Furthermore, the heat shock protein genes hsp70 and hspb1 showed greater upregulation during the gradual thermal exposure compared to the abrupt exposure. Though the differences in response to each thermal regime varied in magnitude among the different populations, the types of responses were very similar (i.e., following the gradual exposure survivorship increased, developmental time showed no effect, and heat shock protein gene upregulation during the exposure increased). Therefore, the enhanced protective effect of the heat shock response during gradual exposures appears to be conserved within the species despite population-level differences in thermal tolerance. Thus, an ecologically relevant thermal exposure likely enables improved cellular protective mechanisms by allowing for an effective and timely heat shock response, which plays a role in mitigating the effects of thermal stress and thereby enhances tolerance to elevated temperatures.


Assuntos
Copépodes/fisiologia , Resposta ao Choque Térmico , Temperatura Alta , Termotolerância , Animais , Organismos Aquáticos/genética , Organismos Aquáticos/fisiologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , California , Copépodes/genética , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP27/genética , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo
20.
Proc Natl Acad Sci U S A ; 116(1): 205-210, 2019 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-30559180

RESUMO

The E3 ubiquitin ligase X-linked inhibitor of apoptosis (XIAP) acts as a molecular rheostat for the immune deficiency (IMD) pathway of the tick Ixodes scapularis How XIAP activates the IMD pathway in response to microbial infection remains ill defined. Here, we identified the XIAP enzymatic substrate p47 as a positive regulator of the I. scapularis IMD network. XIAP polyubiquitylates p47 in a lysine 63-dependent manner and interacts with the p47 ubiquitin-like (UBX) module. p47 also binds to Kenny (IKKγ/NEMO), the regulatory subunit of the inhibitor of nuclear factor (NF)- κB kinase complex. Replacement of the amino acid lysine to arginine within the p47 linker region completely abrogated molecular interactions with Kenny. Furthermore, mitigation of p47 transcription levels through RNA interference in I. scapularis limited Kenny accumulation, reduced phosphorylation of IKKß (IRD5), and impaired cleavage of the NF-κB molecule Relish. Accordingly, disruption of p47 expression increased microbial colonization by the Lyme disease spirochete Borrelia burgdorferi and the rickettsial agent Anaplasma phagocytophilum Collectively, we highlight the importance of ticks for the elucidation of paradigms in arthropod immunology. Manipulating immune signaling cascades within I. scapularis may lead to innovative approaches to reducing the burden of tick-borne diseases.


Assuntos
Ixodes/imunologia , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/fisiologia , Anaplasma , Animais , Proteínas de Artrópodes/metabolismo , Proteínas de Artrópodes/fisiologia , Borrelia burgdorferi , Drosophila , Técnicas de Inativação de Genes , Ixodes/microbiologia , Ixodes/fisiologia , NF-kappa B/metabolismo , Domínios Proteicos , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/metabolismo , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/fisiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA