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1.
Aquat Toxicol ; 276: 107124, 2024 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-39423743

RESUMO

Significant concerns have been raised by the widespread pollutants phoxim (PHO) and nanopolystyrene (NP) in the natural environment. This study evaluated the toxicity effects on the hepatopancreas of Eriocheir sinensis caused by NP and/or PHO at concentrations found in the environment. Subchronic exposure to NP and/or PHO triggered hepatopancreas histological damage within a 21-day exposure period. The NP, PHO, and co-exposure (NPO) groups exhibited fewer blister-like (B) cells, along with the appearance of vacuolation. Furthermore, these exposures induced impairment in the hepatic tubule mucus barrier and mechanical barrier, as evidenced by altered expression of oxidative stress-related genes, mucin-related genes, and TJ-related genes. Additionally, alterations in immunity-related genes and inflammatory cytokine genes expression were observed. The findings showed that hepatopancreas inflammation was caused by both individual and combined exposure to NP and PHO and that the inflammatory response was exacerbated by the co-exposure. The possible pathways of hepatopancreas toxicity were further investigated by transcriptomic analysis. Hepatopancreas inflammation was brought on by subchronic exposure to PHO and co-exposure; this inflammation was exacerbated by co-exposure and was backed by the activation of NF-κB signaling pathway via targeting-related genes. In summary, this research represents the initial documentation, to the best of our understanding of the detrimental effects of exposured to NP and/or PHO at levels found in the environment disrupt the hepatopancreas mucus and mechanical barrier in crustaceans, triggering inflammatory responses. These findings highlight the significance of NP and/or PHO pollution for hepatopancreas health.

2.
Materials (Basel) ; 17(17)2024 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-39274580

RESUMO

Mg-Gd-Y-Zn-Mn (MVWZ842) is a kind of high rare earth magnesium alloy with high strength, high toughness and multi-scale strengthening mechanisms. After heat treatment, the maximum tensile strength of MVWZ842 alloy is more than 550 MPa, and the elongation is more than 5%. Because of its great mechanical properties, MVWZ842 has broad application potential in aerospace and rail transit. However, the addition of high rare earth elements makes the deformation resistance of MVWZ842 alloy increase to some extent. This leads to the difficulty of direct plastic processing forming and large structural part shaping. Friction stir welding (FSW) is a convenient fast solid-state joining technology. When FSW is used to weld MVWZ842 alloy, small workpieces can be joined into a large one to avoid the problem that large workpieces are difficult to form. In this work, a high-quality joint of MVWZ842 alloy was achieved by FSW. The microstructure and properties of this high-strength magnesium alloy after friction stir welding were studied. There was a prominent onion ring characteristic in the nugget zone. After the base was welded, the stacking fault structure precipitated in the grain. There were a lot of broken long period stacking order (LPSO) phases on the retreating side of the nugget zone, which brought the effect of precipitation strengthening. Nano-α-Mn and the broken second phase dispersed in the matrix in the nugget zone, which made the grains refine. A relatively complete dynamic recrystallization occurred in the nugget zone, and the grains were refined. The welding coefficient of the welded joint exceeded 95%, and the hardness of the weld nugget zone was higher than that of the base. There were a series of strengthening mechanisms in the joint, mainly fine grain strengthening, second phase strengthening and solid solution strengthening.

3.
Materials (Basel) ; 17(14)2024 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-39063861

RESUMO

Building formwork is a kind of temporary supporting structure consumable material used in the construction field. In recent years, building formwork has gradually developed to become lighter, more environmentally friendly, and have higher performance. This sets higher requirements for the materials used to make building formwork. There is an urgent need to find a lighter and more durable material for building formwork. Magnesium alloys possess the advantages of low density, high alkali resistance, and high strength. As a building formwork material, it can reduce the weight of formwork and improve its durability. Therefore, a magnesium alloy is considered a material with high potential for building formwork. Currently, magnesium alloy building formwork has attracted the attention of many companies and research and development institutions, with preliminary research applications and good feedback on usage effects. It is highly possible to obtain the opportunity to put it into market application. However, to be applied on a large scale, there are still some important problems that need to be solved. These problems fall into three main areas, including the relatively low processing efficiency of magnesium alloy materials, the unstable price of magnesium alloys, and the fact that the formwork is easily corroded during storage. Firstly, at present, the main processing methods for magnesium alloy building formwork are casting and extrusion, and the production efficiency of both methods needs to be improved. Secondly, high-performance magnesium alloy materials are usually more expensive, which is not conducive to the large-scale application of the formwork. The price of magnesium alloys has fluctuated greatly in recent years, which increases the difficulty of promoting magnesium alloy building formwork. Thirdly, in the atmosphere, the oxide film on the surface of the magnesium alloy cannot play an effective role in corrosion resistance. So, surface treatment is necessary for magnesium alloy building formwork. Among the various surface treatment methods for magnesium alloys, the chemical conversion method has the advantages of being easy to operate, cost-effective, and having good corrosion resistance. It may be a very suitable protective method for large-scale applications of magnesium alloy building formwork and possesses excellent potential for application. The future of magnesium alloy building formwork will focus on new low-cost materials, high-efficiency processing technology, and low-cost green anti-corrosion technology. With in-depth research and the maturation of technology, magnesium alloy formwork is expected to play a more important role in the construction industry.

4.
Fish Shellfish Immunol ; 151: 109626, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38797334

RESUMO

In arthropods, the involvement of Dscam (Down syndrome cell adhesion molecule) in innate immunity has been extensively demonstrated. Its cytoplasmic tail contains multiple conserved functional sites, which indicates its involvement in different intracellular signaling pathways. In this study, we focused on the role of the cytoplasmic tail of Dscam in the Chinese mitten crab (Eriocheir sinensis) immune defense. In the group with cytoplasmic tail knockdown (the site was located on constant exons 37 and 38), 3885 differentially expressed genes (DEGs) were identified. The DEGs were enriched in small molecule binding, protein-containing complex binding, and immunity-related pathways. The expression of selected genes were validated using quantitative real-time reverse transcription PCR. We identified key Cell cycle, Janus kinase (JAK)-signal transducer, activator of transcription (STAT) and mitogen-activated protein kinase (MAPK) signaling pathway genes, the results indicated that the cytoplasmic tail of Dscam controls antibacterial responses by regulating cell proliferation-related genes in hemocytes.


Assuntos
Proteínas de Artrópodes , Braquiúros , Hemócitos , Imunidade Inata , Animais , Braquiúros/genética , Braquiúros/imunologia , Hemócitos/imunologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/química , Imunidade Inata/genética , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Moléculas de Adesão Celular/imunologia , Regulação da Expressão Gênica/imunologia , Proliferação de Células
5.
Materials (Basel) ; 17(6)2024 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-38541439

RESUMO

As the lightest structural metal material, magnesium alloys possess good casting properties, high electrical and thermal conductivity, high electromagnetic shielding, and excellent damping properties. With the increasing demand for lightweight, high-strength, and high-damping structural materials in aviation, automobiles, rail transit, and other industries with serious vibration and noise, damping magnesium alloy materials are becoming one of the important development directions of magnesium alloys. A comprehensive review of the progress in this field is conducive to the development of damping magnesium alloys. This review not only looks back on the traditional damping magnesium alloys represented by Mg-Zr alloys, Mg-Cu-Mn alloys, etc. but also introduces the new damping magnesium materials, such as magnesium matrix composites and porous magnesium. But up to now, there have still been some problems in the research of damping magnesium materials. The effect of spiral dislocation on damping is still unknown and needs to be studied; the contradiction between damping performance and mechanical properties still lacks a good balance method. In the future, the introduction of more diversified damping regulating methods, such as adding other elements and reinforcements, optimizing the manufacturing method of damping magnesium alloy, etc., to solve these issues, will be the development trend of damping magnesium materials.

6.
Ecotoxicol Environ Saf ; 273: 116126, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38387141

RESUMO

Nanopolystyrene (NP) and phoxim (PHO) are common environmental pollutants in aquatic systems. We evaluated the toxic effects of exposure to ambient concentrations of NP and/or PHO in the intestines of the Chinese mitten crab (Eriocheir sinensis). Our study showed that histopathological changes were observed in the intestines. Specifically, NP and/or PHO exposure increased intraepithelial lymphocytes. Furthermore, NP and/or PHO exposure induced oxidative stress, as evidenced by a significant decrease in superoxide dismutase activity (SOD), peroxidase activity (POD), and total antioxidant capacity (T-AOC). Pro-inflammatory gene expression and transcriptome analysis demonstrated that NP and/or PHO exposure induced the intestinal inflammatory response. Transcriptome results showed that NP and/or PHO exposure upregulated the NF-κB signaling pathway, which is considered a key pathway in the inflammatory response. Additionally, the expression of pro-inflammatory genes significantly increased after a single exposure to NP or PHO, but it exhibited a significant decrease after the co-exposure. The downregulation of these genes in the co-exposure group likely suggested that the co-exposure mitigated intestinal inflammation response in E. sinensis. Collectively, our findings mainly showed that NP and/or PHO exposure at ambient concentrations induces oxidative stress and inflammatory response in the intestines of E. sinensis.


Assuntos
Braquiúros , Compostos Organotiofosforados , Estresse Oxidativo , Animais , Antioxidantes/metabolismo , Intestinos , Inflamação/induzido quimicamente , Braquiúros/metabolismo
7.
Clin Exp Metastasis ; 41(2): 81-90, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38396262

RESUMO

OBJECTIVE: Aberrant expression of CD276 has been reported in malignant tumors. However, the exact role and mechanisms of CD276 influence the progression of esophageal squamous cell carcinoma (ESCC) still need to be understood. METHODS: Bioinformatics analysis of data from The Cancer Genome Atlas and Gene Expression Omnibus databases, along with immunohistochemistry staining, was used to explore the expression patterns of CD276 in ESCC. Cell counting kit-8 and Transwell assays were employed to evaluate the effects of CD276 expression on tumor cell proliferation and motility. Western blotting and Transwell assays were used to explore the potential pathways through which CD276 mediates the progression of ESCC. Moreover, the in vivo role of CD276 in tumor progression was investigated by establishing a lung metastasis mouse model. RESULTS: A significant upregulation of CD276 was observed in ESCC tissues compared to adjacent tissues. The inhibition of CD276 had no evident impact on ESCC cell proliferation but notably hindered their migratory and invasive properties and the expression of epithelial-mesenchymal transition (EMT) markers. Inversely, overexpressing CD276 led to an upregulation of EMT markers, underscoring the capacity of CD276 to amplify the motility of ESCC cells. Furthermore, CD276 was found to enhance the migratory and invasive abilities of ESCC cells by activating the TGF-ß/SMAD signaling but not the PI3K/AKT pathway. In vivo studies demonstrated that CD276 facilitates pulmonary metastasis. CONCLUSION: CD276 is significant upregulation in ESCC tissues and facilitates the EMT process in ESCC cells via the TGF-ß/SMAD signaling, thus promoting the progression of ESCC.


Assuntos
Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , Animais , Camundongos , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Transição Epitelial-Mesenquimal , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/patologia , Regulação Neoplásica da Expressão Gênica , Fosfatidilinositol 3-Quinases , Fatores de Transcrição/genética , Fator de Crescimento Transformador beta
8.
Ecotoxicol Environ Saf ; 270: 115936, 2024 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-38183751

RESUMO

Nanopolystyrene (NP) and cadmium (Cd) are ubiquitous contaminants in aquatic systems. The present study aimed to investigate the toxic effects of exposure to ambient concentrations of NP and/or Cd on the intestinal tract of the Chinese mitten crab (Eriocheir sinensis). Exposure to NP and/or Cd induced oxidative stress, as evidenced by a significant increase in lipid peroxide content (LPO), total antioxidant capacity (T-AOC), and peroxidase activity (POD), and significant decreases in superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities in E. sinensis. In addition, exposure to NP and/or Cd imbalanced the homeostasis of the intestinal microbiota, as demonstrated by the significantly increased abundance of Spiroplasma. Transcriptomic and metabolomic analyses were performed to investigate the mechanisms underlying intestinal toxicity. Our results showed that ferroptosis, ABC transporters, phosphotransferase system, apoptosis, and leukocyte transendothelial migration were disturbed after exposure to NP and/or Cd. In particular, Cd exposure affected mucin type O-glycan biosynthesis, purine metabolism, and neuroactive ligand-receptor interaction. Intriguingly, co-exposure to NP and Cd might mitigate intestinal toxicity by decreasing oxidative stress and affecting these pathways. Taken together, our study clearly demonstrates that exposure to NP and/or Cd at environmentally relevant concentrations causes intestinal toxicity in E. sinensis.


Assuntos
Braquiúros , Cádmio , Animais , Cádmio/toxicidade , Antioxidantes/metabolismo , Estresse Oxidativo , Intestinos , Braquiúros/metabolismo
9.
Materials (Basel) ; 16(23)2023 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-38068060

RESUMO

Magnesium alloys with high damping, high specific strength and low density have attracted great attention in recent years. However, the application of magnesium alloys is limited by the balance between their mechanical and damping properties. The strength and plasticity of magnesium alloys with high damping performance often cannot meet the industrial requirements. Understanding the damping mechanism of magnesium alloys is significant for developing new materials with high damping and mechanical properties. In this paper, the damping mechanisms and internal factors of the damping properties of magnesium alloys are comprehensively reviewed. Some damping mechanisms have been studied by many scholars, and it has been found that they can be used to explain damping performance. Among existing damping mechanisms, the G-L dislocation theory, twin damping mechanism and interface damping mechanism are considered common. In addition, some specific long-period stacking ordered (LPSO) phases' crystal structures are conducive to dislocation movement, which is good for improving damping performance. Usually, the damping properties of magnesium alloys are affected by some internal factors directly, such as dislocation density, solute atoms, grain texture and boundaries, etc. These internal factors affect damping performance by influencing the dissipation of energy within the crystal. Scholars are working to find novel damping mechanisms and suitable solute atoms that can improve damping performance. It is important to understand the main damping mechanisms and the internal factors for guiding the development of novel high-damping magnesium alloys.

10.
Dev Comp Immunol ; 140: 104619, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36535491

RESUMO

It has been widely established that Down syndrome cell adhesion molecule (Dscam) regulates arthropod cellular endocytosis. However, the signal transduction pathways and molecular mechanisms of the regulatory process remain unclear. Our previous study identified a Dscam-mediated immune signal transduction pathway that regulates cellular antimicrobial peptide expression, and a conserved endocytosis motif encoded by exon 33 in the cytoplasmic tail of transmembrane Dscam. Therefore, the present study aimed to determine the transcriptional response of the Chinese mitten crab (Eriocheir sinensis) Dscam with a cytoplasmic tail encoded by different exons. In the group of exon 32 knockdown, 306 differentially expressed genes (DEGs) were identified, and 3579 differentially expressed genes (DEGs) were identified in the group of exon 33 knockdown (green fluorescent protein, (GFP) as control). The DEGs were enriched in small molecule binding, protein-containing complex binding, and immunity-related pathways. Quantitative real-time reverse transcription PCR validated the data for selected genes. Our study contributes to the understanding of the immune defense mechanism in E. sinensis and the development of the innate immune system, thus providing insights into disease control and prevention in aquaculture.


Assuntos
Braquiúros , Hemócitos , Animais , Éxons/genética , Fagocitose , Citoesqueleto de Actina/metabolismo , Expressão Gênica , Antibacterianos/metabolismo , Braquiúros/genética , Braquiúros/metabolismo , Regulação da Expressão Gênica , Proteínas de Artrópodes/genética , Imunidade Inata/genética , Filogenia
11.
Materials (Basel) ; 15(22)2022 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-36431657

RESUMO

Designing new materials for vibration and noise reduction that are lightweight is of great significance for industrial development. Magnesium (Mg) alloy is considered one of the best damping metal structural materials because of its low density, high specific strength, good energy storage characteristics and rich resources. Solution atoms have an important effect on the damping capacities of Mg alloys, but the relevant laws have not been completely clarified. In this work, two kinds of alloying elements (Ga and Er) with various atomic sizes were selected to study the metallographic structure and damping capacities of binary Mg-X (X = Ga and Er) alloys in the as-cast and solid solution states, respectively. Solution treatment can improve the damping capacities of binary Mg-X (X = Ga and Er) alloys, and the damping mechanisms of the two solid solution alloys are consistent with the G-L damping mechanism. The influence of alloy elements with different atomic sizes on damping capacities is also different. This influence is due to the various radii of solute atoms and Mg atoms which can result in different degrees of lattice distortion. This work provides a research basis for development and design of high-performance damping Mg alloy materials.

12.
Mol Nutr Food Res ; 66(24): e2200168, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36310136

RESUMO

SCOPE: Proliferation and differentiation of intestinal stem cells (ISCs) are crucial for functional restoration after injury, which can be regulated by nutritional molecules. Aspartate is implicated in maintaining intestinal barrier after injury, but underlying mechanisms remain elusive. Here, this study seeks to investigate if aspartate alleviates colonic epithelial damage by regulating ISC function, and to elucidate its mechanisms. METHODS AND RESULTS: Eight-week-old male C57BL/6 mice supplement with or without 1% L-aspartate are subjected to drinking water or 2.5% DSS to induce colitis. In this study, aspartate administration alleviates the severity of colitis, as indicated by reduced body weight loss, colon shortening, and inhibited pro-inflammatory cytokine expression in DSS-challenged mice. Additionally, aspartate promotes colonic epithelial cell proliferation and differentiation after DSS-induced damage in mice. Pretreatment with aspartate not only enhances ISC proliferation but also induces ISC differentiation toward enterocytes and goblet cells, which prevent TNF-α-induced colonoid damage. Mechanistically, aspartate ameliorates DSS/TNF-α-induced perturbation of mitochondrial metabolism and maintains mitochondrial dynamics in colonic epithelium and colonoids. Moreover, aspartate-mediated ISC proliferation and differentiation are primarily dependent on mitochondrial fusion rather than fission. CONCLUSIONS: The findings indicate that aspartate promotes ISC proliferation and differentiation to alleviate colonic epithelial damage by regulation of mitochondrial metabolism and dynamics.


Assuntos
Ácido Aspártico , Colite , Animais , Masculino , Camundongos , Ácido Aspártico/farmacologia , Diferenciação Celular , Proliferação de Células , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colo/metabolismo , Sulfato de Dextrana/efeitos adversos , Modelos Animais de Doenças , Mucosa Intestinal/metabolismo , Camundongos Endogâmicos C57BL , Dinâmica Mitocondrial , Fator de Necrose Tumoral alfa/metabolismo
13.
Innate Immun ; 27(1): 23-30, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33232194

RESUMO

To test the dynamic changes of the expression of genes and microRNA in the gastrocnemius muscle after LPS challenge, 36 piglets were assigned to a control group (slaughtered 0 h after saline injection) and LPS groups (slaughtered at 1 h, 2 h, 4 h, 8 h, and 12 h after LPS treatment, respectively). After LPS treatment, the mRNA expression of IL-1ß, IL-6, and TNF-α reached maximal levels at 1 h, 2 h, and 1 h, respectively (P < 0.05), and mRNA expression of TLR4, NODs, muscle-specific ring finger 1, and muscle atrophy F-box peaked at 12 h (P < 0.05). Moreover, the expression of miR-122, miR-135a, and miR-370 reduced at 1 h, 1 h, and 2 h, respectively (P < 0.05), and miR-34a, miR-224, miR-132, and miR-145 reached maximum expression levels at 1 h, 1 h, 2 h, and 4 h, respectively (P < 0.05). These results suggested that mRNA expression of pro-inflammatory cytokines was elevated in the early stage, mRNA expression of genes related to TLR4 and NODs signaling pathways and protein degradation increased in the later phase, and the expression of microRNA related to muscle inflammation and protein degradation changed in the early stage after LPS injection.


Assuntos
Regulação da Expressão Gênica/genética , Lipopolissacarídeos/toxicidade , MicroRNAs/biossíntese , MicroRNAs/genética , Proteínas Musculares/genética , Miosite/genética , Animais , Citocinas/biossíntese , Citocinas/genética , Cinética , Masculino , Proteínas Musculares/metabolismo , Miosite/patologia , Proteínas Adaptadoras de Sinalização NOD/genética , Transdução de Sinais/genética , Suínos , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
14.
Br J Nutr ; 123(8): 859-869, 2020 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-31524111

RESUMO

Flaxseed oil is rich in α-linolenic acid (ALA), which is the metabolic precursor of EPA and DHA. The present study investigated the effect of flaxseed oil supplementation on lipopolysaccharide (LPS)-induced muscle atrophy and carbohydrate oxidation impairment in a piglet model. Twenty-four weaned pigs were used in a 2 × 2 factorial experiment including dietary treatment (5 % maize oil v. 5 % flaxseed oil) and LPS challenge (saline v. LPS). On day 21 of treatment, the pigs were injected intraperitoneally with 100 µg/kg body weight LPS or sterile saline. At 4 h after injection, blood, gastrocnemius muscle and longissimus dorsi muscle were collected. Flaxseed oil supplementation increased ALA, EPA, total n-3 PUFA contents, protein:DNA ratio and pyruvate dehydrogenase complex quantity in muscles (P < 0·05). In addition, flaxseed oil reduced mRNA expression of toll-like receptor (TLR) 4 and nucleotide-binding oligomerisation domain protein (NOD) 2 and their downstream signalling molecules in muscles and decreased plasma concentrations of TNF-α, IL-6 and IL-8, and mRNA expression of TNF-α, IL-1ß and IL-6 (P < 0·05). Moreover, flaxseed oil inclusion increased the ratios of phosphorylated protein kinase B (Akt) 1:total Akt1 and phosphorylated Forkhead box O (FOXO) 1:total FOXO1 and reduced mRNA expression of FOXO1, muscle RING finger (MuRF) 1 and pyruvate dehydrogenase kinase 4 in muscles (P < 0·05). These results suggest that flaxseed oil might have a positive effect on alleviating muscle protein loss and carbohydrates oxidation impairment induced by LPS challenge through regulation of the TLR4/NOD and Akt/FOXO signalling pathways.


Assuntos
Óleo de Semente do Linho/farmacologia , Lipopolissacarídeos/toxicidade , Proteínas Musculares/metabolismo , Músculo Esquelético/efeitos dos fármacos , Suínos , Animais , Metabolismo dos Carboidratos , Ácidos Graxos/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas Musculares/genética , Oxirredução , Fosforilação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
15.
J Biol Chem ; 294(44): 16440-16450, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31537644

RESUMO

Alternatively-spliced hypervariable immunoglobulin domain-encoding molecules, called Down syndrome cell adhesion molecule (Dscam), have been widely detected as components of the arthropod immune system. Although its ability to specifically bind pathogens and enable phagocytosis of bacteria has been elucidated, the signal transduction mechanisms or effectors that activate post-Dscam-binding pathogens remain poorly characterized. Here, we reveal the alternative splicing exons of Dscam's cytoplasmic tail and its isoforms in the hemocytes of crab (Eriocheir sinensis), showing that the expression of Dscam was acutely induced after an immune challenge, which suggested its functioning for innate immunity. Significantly decreased expression levels of antimicrobial molecular peptides (AMPs) were detected in Dscam-silenced crab hemocytes in vitro, which coincided with their vulnerability to infection by Staphylococcus aureus and higher bacterial concentrations occurring in Dscam-silenced crabs in vivo Further experimental investigation demonstrated that Dscam-regulated AMP expression via the Src homology (SH)3-binding domain in the first constant exon translated protein of the cytoplasmic tail bound with the SH3 domain of the Dock, an SH3/SH2 adaptor protein required for axon guidance. Dock promoted extracellular signal-regulated kinase (ERK) phosphorylation via indirect binding and then regulated dorsal phosphorylation and translocation from the cytoplasm to the nucleus, subsequently promoting AMP expression for the effective removal of bacteria. To the best of our knowledge, this comprehensive study is the first to highlight the critical role of the alternatively-spliced Dscam cytoplasmic tail in antimicrobial control activity. It also suggests possible cross-talk occurring between Dscam and other pattern recognition receptors.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Moléculas de Adesão Celular/metabolismo , Imunidade Inata/fisiologia , Processamento Alternativo/genética , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/metabolismo , Moléculas de Adesão Celular/genética , Crustáceos/metabolismo , Éxons , Hemócitos/metabolismo , Imunidade Inata/genética , Fagocitose , Filogenia , Isoformas de Proteínas/metabolismo , Splicing de RNA/genética , Alinhamento de Sequência , Proteínas de Xenopus/genética , Proteínas de Xenopus/metabolismo
16.
Fish Shellfish Immunol ; 87: 460-469, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30685464

RESUMO

B52 is a member of the classical serine/arginine (SR)-rich proteins, which are phylogenetically conserved and play significant roles in mRNA maturation, including alternative splicing. In the present study, the docking site, selector sequences and locus control region of the Chinese mitten crab (Eriocheir sinensis) Down syndrome cell adhesion molecule (EsDscam) were identified. Alternative splicing of Dscam is essential to generate different isoforms. We also isolated and characterised the B52 gene from E. sinensis (EsB52). The 876 bp open reading frame of EsB52 encodes a 291 amino acid residue polypeptide, and EsB52 has two RNA recognition motifs (RRMs) at the N-terminus and an arginine/serine-rich domain at the C-terminus. Each RRM contains two degenerate short submotifs, RNP-1 and RNP2. Analysis of tissue distribution revealed that EsB52 mRNA expression was widespread in all tested tissues, and especially high in brain and hemocytes. In hemocytes, EsB52 was upregulated significantly after stimulation with pathogen-associated molecular patterns and bacteria. Furthermore, EsB52 RNAi decreased the number of Ig7 inclusion in mRNA rather than Ig2 or Ig3. Taken together, these findings suggest that EsB52 acts as an alternative splicing activator of EsDscam.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Moléculas de Adesão Celular/genética , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Fatores de Processamento de Serina-Arginina/genética , Fatores de Processamento de Serina-Arginina/imunologia , Processamento Alternativo , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Moléculas de Adesão Celular/metabolismo , Feminino , Perfilação da Expressão Gênica , Masculino , Filogenia , Alinhamento de Sequência , Fatores de Processamento de Serina-Arginina/química
17.
Fish Shellfish Immunol ; 84: 491-501, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30253180

RESUMO

The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signalling pathway plays a significant role in immune responses to pathogens. In invertebrates, three core components (Domeless, Hopscotch and STAT92E) of the JAK/STAT pathway were first identified in Drosophila melanogaster. In the present study, we report the cloning and characterisation of DOME, JAK and STAT from Eriocheir sinensis. EsDOME, EsJAK and EsSTAT are 1299, 1110 and 794 amino acid proteins encoded by 4200, 3333 and 2385 bp open reading frames, respectively. Bioinformatics analysis revealed that all three share specific domains with corresponding functions. Quantitative real-time PCR showed that all three components were highly expressed in various tissues including the gill, hepatopancreas and hemocytes. Moreover, EsDOME, EsJAK and EsSTAT were upregulated significantly in hemocytes after bacterial challenge. In addition, the JAK/STAT signalling pathway positively regulates antimicrobial peptide expression during the antibacterial immune response. Thus, the JAK/STAT pathway plays a critical role in bacterial immune responses in E. sinensis.


Assuntos
Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Braquiúros/genética , Braquiúros/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Transdução de Sinais/genética , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Sequência de Bases , Perfilação da Expressão Gênica , Filogenia , Distribuição Aleatória , Alinhamento de Sequência , Staphylococcus aureus/fisiologia , Vibrio parahaemolyticus/fisiologia
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