RESUMO
Posttranslational modifications expand the functions of immune-related proteins, especially during infections. The respiratory glycoprotein, hemocyanin, has been implicated in many other functions, but the role of phosphorylation modification in its functional diversity is not fully understood. In this study, we show that Penaeus vannamei hemocyanin (PvHMC) undergoes phosphorylation modification during bacterial infection. Dephosphorylation of PvHMC mediated by P. vannamei protein phosphatase 2A catalytic increases its in vitro antibacterial activity, whereas phosphorylation by P. vannamei casein kinase 2 catalytic subunit α decreases its oxygen-carrying capacity and attenuates its in vitro antibacterial activity. Mechanistically, we show that Thr517 is a critical phosphorylation modification site on PvHMC to modulate its functions, which when mutated attenuates the action of P. vannamei casein kinase 2 catalytic subunit α and P. vannamei protein phosphatase 2A catalytic, and hence abolishes the antibacterial activity of PvHMC. Our results reveal that phosphorylation of PvHMC modulates its antimicrobial functions in penaeid shrimp.
Assuntos
Hemocianinas , Penaeidae , Animais , Hemocianinas/metabolismo , Penaeidae/metabolismo , Caseína Quinase II/metabolismo , Proteína Fosfatase 2/metabolismo , Antibacterianos/farmacologia , Antibacterianos/metabolismoRESUMO
Ferroptosis, characterized by iron-dependent cell death, has recently emerged as a critical defense mechanism against microbial infections. The present study aims to investigate the involvement of exosomes in the induction of ferroptosis and the inhibition of bacterial infection in crustaceans. Our findings provide compelling evidence for the pivotal role of exosomes in the immune response of crustaceans, wherein they facilitate intracellular iron accumulation and activate the ferroptotic pathways. Using RNA-seq and bioinformatic analysis, we demonstrate that cytochrome P450 (CYP) can effectively trigger ferroptosis. Moreover, by conducting an analysis of exosome cargo proteins, we have identified the participation of six-transmembrane epithelial antigen of prostate 4 in the regulation of hemocyte ferroptotic sensitivity. Subsequent functional investigations unveil that six-transmembrane epithelial antigen of prostate 4 enhances cellular Fe2+ levels, thereby triggering Fenton reactions and accelerating CYP-mediated lipid peroxidation, ultimately culminating in ferroptotic cell death. Additionally, the Fe2+-dependent CYP catalyzes the conversion of arachidonic acid into 20-hydroxyeicosatetraenoic acid, which activates the peroxisome proliferator-activated receptor. Consequently, the downstream target of peroxisome proliferator-activated receptor, cluster of differentiation 36, promotes intracellular fatty acid accumulation, lipid peroxidation, and ferroptosis. These significant findings shed light on the immune defense mechanisms employed by crustaceans and provide potential strategies for combating bacterial infections in this species.
Assuntos
Bactérias , Crustáceos , Exossomos , Ferroptose , Ferro , Sistema Enzimático do Citocromo P-450/metabolismo , Exossomos/metabolismo , Ferroptose/fisiologia , Ferro/metabolismo , Peroxidação de Lipídeos , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Oxirredutases/metabolismo , Proteínas de Membrana/metabolismo , Antígenos CD36/metabolismo , RNA-Seq , Compostos Ferrosos/metabolismo , Crustáceos/citologia , Crustáceos/genética , Crustáceos/metabolismo , Crustáceos/microbiologia , Ácidos Hidroxieicosatetraenoicos , Ácido Araquidônico/metabolismo , Ácidos Graxos/metabolismo , Bactérias/metabolismoRESUMO
Crustaceans such as shrimps and crabs, hold significant ecological significance and substantial economic value within marine ecosystems. However, their susceptibility to disease outbreaks and pathogenic infections has posed major challenges to production in recent decades. As invertebrate, crustaceans primarily rely on their innate immune system for defense, lacking the adaptive immune system found in vertebrates. Mucosal immunity, acting as the frontline defense against a myriad of pathogenic microorganisms, is a crucial aspect of their immune repertoire. This review synthesizes insights from comparative immunology, highlighting parallels between mucosal immunity in vertebrates and innate immune mechanisms in invertebrates. Despite lacking classical adaptive immunity, invertebrates, including crustaceans, exhibit immune memory and rely on inherent "innate immunity factors" to combat invading pathogens. Drawing on parallels from mammalian and piscine systems, this paper meticulously explores the complex role of mucosal immunity in regulating immune responses in crustaceans. Through the extrapolation from well-studied models like mammals and fish, this review infers the potential mechanisms of mucosal immunity in crustaceans and provides insights for research on mucosal immunity in crustaceans.
Assuntos
Crustáceos , Imunidade nas Mucosas , Animais , Crustáceos/imunologia , Imunidade InataRESUMO
Hemocyanin is a multifunctional protein present in arthropods and mollusks, responsible for oxygen transport and participating in multiple roles of immune defense including antibacterial activity. However, the molecular basis of how hemocyanin recognizes pathogens and exerts antibacterial activity remains poorly understood. In the present study, the pull-down assay was used to isolate Vibrio parahaemolyticus outer membrane proteins (OMPs) that bind to Litopenaeus vannamei hemocyanin. Two interacting OMPs bands were determined as OmpC and OmpU, and the heterogeneous interaction between hemocyanin and the two OMPs was further confirmed by far-Western blot. After construction of ompC and ompU deletion mutants, we found that the agglutinating activity and antibacterial activity of hemocyanin significantly decreased compared to the wild-type strain. After hemocyanin treatment, we identified four intracellular proteins of V. parahaemolyticus, including fructose-bisphosphate aldolase and ribosomal proteins could interact with rOmpC and rOmpU, respectively. Furthermore, we found that the mRNA levels of ompC, ompU, fbaA, rpsB and rpsC significantly decreased after hemocyanin treatment. These findings indicated that OmpC and OmpU are the key targets for L. vannamei hemocyanin recognize pathogens and exert its antibacterial activity.
Assuntos
Penaeidae , Vibrio parahaemolyticus , Animais , Hemocianinas , Sequência de Aminoácidos , AntibacterianosRESUMO
Succinate dehydrogenase (SDH) is a crucial enzyme in the tricarboxylic acid cycle (TCA) and has established roles in immune function. However, the understanding of SDH in Penaeus vannamei, particularly its involvement in immune responses, is currently limited. Through affinity proteomics, a potential interaction between hemocyanin (HMC) and SDH in shrimp has been identified. The successful cloning of PvSDH in this study has revealed a high degree of evolutionary conservation. Additionally, it has been found that hemocyanin regulates SDH not only at the transcriptional and enzymatic levels but also through confirmed protein-protein interactions observed via Co-immunoprecipitation (CoIP) assay. Moreover, by combining PvHMC knockdown and Vibrio parahaemolyticus challenge, it was demonstrated that fumaric acid, a product of SDH, enhances the host's immune resistance to pathogen infection by modulating the expression of antimicrobial peptides. This research provides new insights into HMC as a crucial regulator of SDH, potentially impacting glycometabolism and the dynamics of immune responses.
Assuntos
Proteínas de Artrópodes , Hemocianinas , Penaeidae , Succinato Desidrogenase , Vibrio parahaemolyticus , Animais , Penaeidae/imunologia , Penaeidae/genética , Hemocianinas/imunologia , Hemocianinas/genética , Hemocianinas/metabolismo , Vibrio parahaemolyticus/fisiologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/metabolismo , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismo , Succinato Desidrogenase/imunologia , Glucose/metabolismo , Imunidade Inata/genética , Regulação da Expressão Gênica/imunologia , Sequência de Aminoácidos , Filogenia , Alinhamento de SequênciaRESUMO
Hemocyanin is the main respiratory protein of arthropods and is formed by hexameric and/or oligomeric subunits. Due to changes in the living environment and gene rearrangement, various hemocyanin subtypes and subunits evolved in crustaceans. This paper reviews the various hemocyanin subtypes and isoforms in shrimp and analyses published genomic data of sixteen hemocyanin family genes from Litopenaeus vannamei to explore the evolution of hemocyanin genes, subunits, and protein structure. Analysis of hemocyanin subtypes distribution and structure in various tissues was also performed and related to multiple and tissue-specific functions, i.e., immunological activity, immune signaling, phenoloxidase activity, modulation of microbiota homeostasis, and energy metabolism. The functional diversity of shrimp hemocyanin due to molecular polymorphism, transcriptional regulation, alternative splicing, degradation into functional peptides, interaction with other proteins or genes, and structural differences will also be highlighted for future research. Inferences would be drawn from other crustaceans to explain how evolution has changed the structure-function of hemocyanin and its implication for evolutionary research into the multifunctionality of hemocyanin and other related proteins in shrimp.
Assuntos
Hemocianinas , Penaeidae , Animais , Isoformas de Proteínas/genética , Peptídeos/genética , Processamento AlternativoRESUMO
Exosomes, secreted by most cells, are critical antimicrobial immune factors in animals. Recent studies of certain key regulators of vesicular transport, the Rab GTPases, have linked Rab dysfunction to regulation of innate immune signaling. However, the relationship between exosomes and Rab GTPases, resulting in antimicrobial activity in vertebrates and invertebrates during pathogenic infection, has not been addressed. In this study, SpRab11a was reported to have a protective effect on the survival rate of mud crabs Scylla paramamosain after Vibrio parahaemolyticus challenge through the stimulation of exosome secretion and modulation of anti-LPS factor (ALF) expression. Furthermore, Sp14-3-3 was confirmed to be densely packaged in exosomes after V. parahaemolyticus infection, which could recruit the MyD88 and TLR by binding the Toll/IL-1R domain to the plasma membrane, promoting the translocation of Dorsal from the cytoplasm into the nucleus, and thereby regulating ALFs expression in the hemocytes of mud crab in response to the bacterial infection. The findings therefore provide, to our knowledge, a novel mechanism that underlies the cross-talk between SpRab11a-regulated exosome formation and ALFs expression in innate immune response in invertebrates, with a crustacean species, mud crab S. paramamosain, as a model study.
Assuntos
Infecções Bacterianas , Braquiúros , Exossomos , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Proteínas de Artrópodes/genética , Exossomos/metabolismo , Imunidade Inata , Filogenia , Proteínas rab de Ligação ao GTP/metabolismoRESUMO
Although invertebrates' innate immunity relies on several immune-like molecules, the diversity of these molecules and their immune response mechanisms are not well understood. Here, we show that Penaeus vannamei hemocyanin (PvHMC) undergoes specific deacetylation under Vibrio parahaemolyticus and LPS challenge. In vitro deacetylation of PvHMC increases its binding capacity with LPS and antibacterial activity against Gram-negative bacteria. Lysine residues K481 and K484 on the Ig-like domain of PvHMC are the main acetylation sites modulated by the acetyltransferase TIP60 and deacetylase HDAC3. Deacetylation of PvHMC on K481 and K484 allows PvHMC to form a positively charged binding pocket that interacts directly with LPS, whereas acetylation abrogates the positive charge to decrease PvHMC-LPS attraction. Besides, V. parahaemolyticus and LPS challenge increases the expression of Pvhdac3 to induce PvHMC deacetylation. This work indicates that, during bacterial infections, deacetylation of hemocyanin is crucial for binding with LPS to clear Gram-negative bacteria in crustaceans.
Assuntos
Hemocianinas , Penaeidae , Animais , Proteínas de Artrópodes/metabolismo , Lipopolissacarídeos/metabolismo , Imunidade Inata , Bactérias Gram-Negativas , Antibacterianos/metabolismoRESUMO
BACKGROUND: The purpose of this study was to investigate the photoprotection effect of peroxiredoxin 1 (PRDX1) protein in ultraviolet B (UVB) irradiation-induced damage of retinal pigment epithelium (RPE) and its possible molecular mechanism. METHODS: ARPE-19 cell viability and apoptosis were assessed by MTT assay and flow cytometry, respectively. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the PRDX1 expression. The corresponding kits were employed to measure the levels or activities of lactate dehydrogenase (LDH), 8-hydroxy-2-deoxyguanosine (8-OHdG), reactive oxygen species (ROS), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD). Western blotting was applied to examine PRDX1 expression and mitogen-activated protein kinase (MAPK) signaling pathway-related proteins. RESULTS: After exposure to 20 mJ/cm2 intensity of UVB irradiation for 24 h, ARPE-19 cells viability was decreased, the leakage degree of LDH and 8-OHdG were increased, and cell apoptosis was elevated. The expression of PRDX1 was significantly down-regulated in UVB-induced ARPE-19 cells. The low expression of PRDX1 was involved in high irradiation intensity. Overexpression of PRDX1 increased cell activity, decreased cell apoptosis, and LDH as well as 8-OHdG leakage in UVB-induced ARPE-19 cells. In addition to alleviating UVB-induced cell damage, PRDX1 overexpression also inhibited UVB-induced oxidative stress (down-regulation of ROS and MDA levels, up-regulation of GSH-Px and SOD activities) and the activation of MAPK signaling pathway in ARPE-19 cells. CONCLUSION: PRDX1 exerts a photoprotection effect on RPE by attenuating UVB-induced cell damage and inhibiting oxidative stress, which can be attributed to the inhibition of MAPK signaling pathway activation.
Assuntos
Apoptose , Sobrevivência Celular , Estresse Oxidativo , Peroxirredoxinas , Espécies Reativas de Oxigênio , Epitélio Pigmentado da Retina , Raios Ultravioleta , Humanos , Epitélio Pigmentado da Retina/efeitos da radiação , Epitélio Pigmentado da Retina/metabolismo , Peroxirredoxinas/metabolismo , Raios Ultravioleta/efeitos adversos , Espécies Reativas de Oxigênio/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Linhagem Celular , Western Blotting , Células Cultivadas , 8-Hidroxi-2'-Desoxiguanosina/metabolismo , Transdução de SinaisRESUMO
As the usage of long-chain perfluoroalkyl and polyfluoroalkyl substances (PFASs) may be gradually restricted, short-chain and even ultra-short-chain PFASs have been widely produced and used, which has put forward new requirements for the simultaneous analysis of the above substances. Using solid phase extraction two-fraction elution and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), an experimental method was established for the simultaneous analysis of ultrashort-chain, short-chain, and long-chain PFASs and the precursor perfluorohexanesulfonamide (FHxSA) in low-concentration water, such as tap water and bottled water. By optimizing the volume of methanol in the first-fraction elution, the concentration of ammonia in the second-fraction elution, and the concentration of ammonium acetate in the mobile phase, the high recovery and low detection limit (0.01-3 ng/L) were obtained. In addition, this method was used to measure nine tap water samples and six bottled water samples for validation, and the results showed that the concentration of PFASs in bottled water was lower than that in tap water. This study first reported the trifluoroacetic acid concentration in bottled water (6.61 ± 9.60 ng/L), which was lower than that in tap water (1712 ± 174 ng/L). The main substances in tap water and bottled water are both ultrashort-chain PFASs (C2-C3), accounting for more than 50%. There are few reports on the simultaneous analysis of ultrashort-chain, short-chain, and long-chain PFASs (C2-C18) and the precursor FHxSA in low-concentration water samples, and the new method can be further developed for different environmental media.
Assuntos
Água Potável , Fluorocarbonos , Poluentes Químicos da Água , Água Potável/análise , Cromatografia Líquida , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas em Tandem/métodos , Poluentes Químicos da Água/análise , Fluorocarbonos/análise , Extração em Fase Sólida/métodosRESUMO
p53, the guardian of the genome, is a short-lived protein that is tightly controlled at low levels by constant ubiquitination and proteasomal degradation in higher organisms. p53 stabilization and activation are early crucial events to cope with external stimuli in cells. However, the role of p53 ubiquitination and its relevant molecular mechanisms have not been addressed in invertebrates. In this study, our findings revealed that both HUWE1 (HECT, UBA, and WWE domain-containing E3 ubiquitin-protein ligase 1) and TRAF6 (tumor necrosis factor receptor-associated factor 6) could serve as E3 ubiquitin ligases for p53 in mud crabs (Scylla paramamosain). Moreover, the expression of HUWE1 and TRAF6 was significantly downregulated during white spot syndrome virus (WSSV) infection, and therefore the ubiquitination of p53 was interrupted, leading to the activation of apoptosis and reactive oxygen species (ROS) signals through p53 accumulation, which eventually suppressed viral invasion in the mud crabs. To the best of our knowledge, this is the first study to reveal the p53 ubiquitination simultaneously induced by two E3 ligases in arthropods, which provides a novel molecular mechanism of invertebrates for resistance to viral infection. IMPORTANCE p53, which is a well-known tumor suppressor that has been widely studied in higher animals, has been reported to be tightly controlled at low levels by ubiquitin-dependent proteasomal degradation. However, recent p53 ubiquitination-relevant research mainly involved an individual E3 ubiquitin ligase, but not whether there exist other mechanisms that need to be explored. The results of this study show that HUWE1 and TRAF6 could serve as p53 E3 ubiquitin ligases and synchronously mediate p53 ubiquitination in mud crabs (Scylla paramamosain), which confirmed the diversity of the p53 ubiquitination regulatory pathway. In addition, the effects of p53 ubiquitination are mainly focused on tumorigenesis, but a few are focused on the host immune defense in invertebrates. Our findings reveal that p53 ubiquitination could affect ROS and apoptosis signals to cope with WSSV infection in mud crabs, which is the first clarification of the immunologic functions and mechanisms of p53 ubiquitination in invertebrates.
Assuntos
Braquiúros , Fator 6 Associado a Receptor de TNF , Proteína Supressora de Tumor p53 , Ubiquitina-Proteína Ligases , Viroses , Vírus da Síndrome da Mancha Branca 1 , Animais , Braquiúros/virologia , Espécies Reativas de Oxigênio/metabolismo , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitinação , Viroses/veterináriaRESUMO
It is well known that exosomes could serve as anti-microbial immune factors in animals. However, despite growing evidences have shown that the homeostasis of the hemolymph microbiota was vital for immune regulation in crustaceans, the relationship between exosomes and hemolymph microbiota homeostasis during pathogenic bacteria infection has not been addressed. Here, we reported that exosomes released from Vibrio parahaemolyticus-infected mud crabs (Scylla paramamosain) could help to maintain the homeostasis of hemolymph microbiota and have a protective effect on the mortality of the host during the infection process. We further confirmed that miR-224 was densely packaged in these exosomes, resulting in the suppression of HSP70 and disruption of the HSP70-TRAF6 complex, then the released TRAF6 further interacted with Ecsit to regulate the production of mitochondrial ROS (mROS) and the expression of Anti-lipopolysaccharide factors (ALFs) in recipient hemocytes, which eventually affected hemolymph microbiota homeostasis in response to the pathogenic bacteria infection in mud crab. To the best of our knowledge, this is the first document that reports the role of exosome in the hemolymph microbiota homeostasis modulation during pathogen infection, which reveals the crosstalk between exosomal miRNAs and innate immune response in crustaceans.
Assuntos
Proteínas de Artrópodes/metabolismo , Braquiúros/imunologia , Exossomos/genética , Hemolinfa/imunologia , Imunidade Inata/imunologia , MicroRNAs/genética , Vibrioses/imunologia , Animais , Proteínas de Artrópodes/genética , Braquiúros/microbiologia , Perfilação da Expressão Gênica , Hemócitos/imunologia , Hemócitos/metabolismo , Hemócitos/microbiologia , Hemolinfa/metabolismo , Hemolinfa/microbiologia , Homeostase , Microbiota , Filogenia , Vibrioses/microbiologia , Vibrio parahaemolyticus/fisiologiaRESUMO
The cellular transcription factors are known to play important roles in virus infection. The present study cloned and characterized a transcription factor CCAAT/Enhancer-binding protein homolog from the shrimp Penaeus vannamei (designates as PvCEBP), and explored its potential functions in white spot syndrome virus (WSSV) infection. PvCEBP has an open reading frame (ORF) of 864 bp encoding a putative protein of 287 amino acids, which contained a highly C-terminal conserved bZIP domain. Phylogenetic tree analysis showed that PvCEBP was evolutionarily clustered with invertebrate CEBPs and closely related to the CEBP of Homarus americanus. Quantitative real-time PCR (qPCR) analysis revealed that PvCEBP was expressed in all examined shrimp tissues, with transcript levels increased in shrimp hemocytes and gill upon WSSV challenge. Furthermore, knockdown of PvCEBP mediated by RNA interference significantly decreased the expression of WSSV genes and viral loads, while enhanced the shrimp survival rate under WSSV challenge. In silico prediction and reporter gene assays demonstrated that PvCEBP could activate the promoter activity of the viral immediate-early gene ie1. Collectively, our findings suggest that PvCEBP is annexed by WSSV to promote its propagation by enhancing the expression of viral immediate-early genes.
Assuntos
Penaeidae , Vírus da Síndrome da Mancha Branca 1 , Animais , Fatores de Transcrição/genética , Penaeidae/genética , Vírus da Síndrome da Mancha Branca 1/fisiologia , Filogenia , Sequência de Aminoácidos , Proteínas de Artrópodes/genéticaRESUMO
The dietary supplementation of red seaweed-derived polysaccharides has been shown to be beneficial to fish and shellfish aquaculture. However, the function of red seaweed (Gracilaria lemaneiformis)-extracted polysaccharide (GLP) on the health status of rabbitfish (Siganus canaliculatus) is still unknown. This study explored the influences of GLP on growth performance, antioxidant activity, and immunity of rabbitfish. Herein, the fish were fed commercial pelleted feed incorporated with the diverse amount of GLP: 0 (control), 0.10 (GLP0.10), and 0.15 g kg-1 (GLP0.15) for 60 days. The results demonstrated that dietary GLP0.15 significantly elevated FBW and WG, while feed utilization efficiency improved (reduced feed conversion ratio and increased protein efficiency ratio) upon GLP0.10 treatment, regarding the control (P < 0.05). Also, dietary administration of GLP0.15 suggestively improved the serum acid phosphatase and lysozyme activity as well as hepatic total antioxidant capacity, catalase, and superoxide dismutase activity. In contrast, GLP0.15decreased the serum alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and malonaldehyde activity when compared to the control (P<0.05). Moreover, the lipase (36.08 and 16.46 U/mgprot in GLP0.10 and GLP0.15, respectively) and amylase (0.43 and 0.23 U/mgprot in GLP0.10 and GLP0.15, respectively) activity recorded the maximum values than the control (8.61 and 0.13 U/mgprot, respectively).Further, the intestinal morphometry was developed (such as increased villus length, width, and area) in the fish fed with a GLP-supplemented diet compared to the control. The KEGG pathway analysis unveiled that several differentially expressed genes (DEGs) in control vs. GLP0.10 and control vs. GLP0.15 were associated with metabolic or immune-associated pathways like antigen processing and presentation, phagosome, complement and coagulation cascades, and platelet activation. The DEGs, namely C3, f5, fgb, MHC1, and cfb, were evaluated in control vs. GLP0.10 and C3 and MHC1 in control vs. GLP0.15, suggesting their possible contributions to GLP-regulated immunity. Additionally, the cumulative mortality of rabbitfish after the Vibrio parahaemolyticus challenge was lower in both GLP0.10 (8.88%) and GLP0.15 (11.11%) than in control (33.33%) (P<0.05). Thus, these findings direct the potential use of GLP as an immunostimulant and growth promoter in rabbitfish aquaculture.
Assuntos
Gracilaria , Alga Marinha , Animais , Antioxidantes/metabolismo , Sulfatos/farmacologia , Imunidade Inata/genética , Suplementos Nutricionais/análise , Dieta/veterinária , Peixes/metabolismo , Polissacarídeos/farmacologia , Ração Animal/análiseRESUMO
Aquatic organisms have to produce proteins or factors that help maintain a stable relationship with microbiota and prevent colonization by pathogenic microorganisms. In crustaceans and other aquatic invertebrates, relatively few of these host factors have been characterized. In this study, we show that the respiratory glycoprotein hemocyanin is a crucial host factor that modulates microbial composition and diversity in the hepatopancreas of penaeid shrimp. Diseased penaeid shrimp (Penaeus vannamei), had an empty gastrointestinal tract with atrophied hepatopancreas, expressed low hemocyanin, and high total bacterial abundance, with Vibrio as the dominant bacteria. Similarly, shrimp depleted of hemocyanin had mitochondrial depolarization, increased reactive oxygen species (ROS) levels, and dysregulation of several energy metabolism-related genes. Hemocyanin silencing together with ROS scavenger (N-acetylcysteine) treatment improved microbial diversity and decreased Vibrio dominance in the hepatopancreas. However, fecal microbiota transplantation after hemocyanin knockdown could not restore the microbial composition in the hepatopancreas. Collectively, our data provide, to our knowledge, new insight into the pivotal role of hemocyanin in modulating microbial composition in penaeid shrimp hepatopancreas via its effect on mitochondrial integrity, energy metabolism, and ROS production.
Assuntos
Hemocianinas/metabolismo , Hepatopâncreas/metabolismo , Penaeidae/microbiologia , Animais , Metabolismo Energético , Hemocianinas/imunologia , Hepatopâncreas/imunologia , Penaeidae/imunologia , Penaeidae/metabolismoRESUMO
Many environmental and pathogenic insults induce endoplasmic reticulum (ER) stress in animals, especially in aquatic ecosystems, where these factors are crucial for life. In penaeid shrimp, pathogens and environmental stressors induce hemocyanin expression, but the involvement of hemocyanin in ER stress response is unknown. We demonstrate that in response to pathogenic bacteria (Vibrio parahaemolyticus and Streptococcus iniae), hemocyanin, ER stress proteins (Bip, Xbp1s, and Chop), and sterol regulatory element binding protein (SREBP) are induced to alter fatty acid levels in Penaeus vannamei. Interestingly, hemocyanin interacts with ER stress proteins to modulate SREBP expression, while ER stress inhibition with 4-Phenylbutyric acid or hemocyanin knockdown attenuates the expression of ER stress proteins, SREBP, and fatty acid levels. Contrarily, hemocyanin knockdown followed by tunicamycin treatment (ER stress activator) increased their expression. Thus, hemocyanin mediates ER stress during pathogen challenge, which consequently modulates SREBP to regulate the expression of downstream lipogenic genes and fatty acid levels. Our findings reveal a novel mechanism employed by penaeid shrimp to counteract pathogen-induced ER stress.
Assuntos
Penaeidae , Proteínas de Ligação a Elemento Regulador de Esterol , Animais , Hemocianinas/genética , Hemocianinas/metabolismo , Penaeidae/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Ecossistema , Estresse do Retículo Endoplasmático , Ácidos Graxos/metabolismo , Bactérias/metabolismo , Proteínas de Choque Térmico/metabolismoRESUMO
Penaeus vannamei is one of the most popular aquaculture species in the world. This species is featured with its fast-growing and delicious taste, which drives people develop various strains. During this process identification of trait-associated markers could effectively increase breeding efficiency. Driven by this, we tried to screen fast-growing key regulators via a FACS-based high throughput method, in which 2-NBDG was applied as a fluorescent indicator for direct glucose uptake measurement. Totally six candidate genes were screened out followed by in vitro validation in 293T cells. After that, the correlation between these genes and shrimp growing was further verified in a hybrid lineage. The expression level of two genes including ATP synthase subunit e and inhibitor of apoptosis protein showed some correlation with shrimp growth speed. Furthermore, we tested these two candidate markers in various lineages and confirmed that ATP synthase subunit e could be a shrimp growth-associated breeding marker.
Assuntos
Penaeidae , Trifosfato de Adenosina , Animais , Biomarcadores , Cruzamento , Humanos , Penaeidae/genética , FenótipoRESUMO
Domestic waste classification provides a new idea for the low-carbon transformation of the cement industry in the context of carbon peaking and carbon neutrality. This study adopted the life cycle assessment to evaluate the environmental feasibility of classified domestic waste as alternative raw material and fuel for clinker production. An area in Shandong Province was selected to investigate the impact of waste transportation. A roadmap was formulated and the calorific value of the refuse-derived fuel using classified domestic waste (RDFCDW) was calculated as 18,977.76 kJ/kg. Compared with conventional waste disposal, disposal in cement kiln had the lowest environmental impact, except for human toxicity potential (HTP). The overall environmental burden of co-production was 7.86% lower than that of conventional production, in which the impact of waste transportation was negligible. Combined with the increasing proportion of clean power applications in 2030 and 2060, the HTP could be decreased by 56.29% and 83.38%, respectively. Accordingly, cement co-production using classified domestic waste as alternative raw material and fuel is a nationally appropriate and environmentally friendly approach.
Assuntos
Eliminação de Resíduos , Humanos , Animais , Estudos de Viabilidade , Meio Ambiente , Carbono , Estágios do Ciclo de Vida , Resíduos Sólidos/análiseRESUMO
MicroRNAs are regulatory molecules that can be packaged into exosomes to modulate cellular response of recipients. While the role of exosomes during viral infection is beginning to be appreciated, the involvement of exosomal miRNAs in immunoregulation in invertebrates has not been addressed. Here, we observed that exosomes released from WSSV-injected mud crabs could suppress viral replication by inducing apoptosis of hemocytes. Besides, miR-137 and miR-7847 were found to be less packaged in mud crab exosomes during viral infection, with both miR-137 and miR-7847 shown to negatively regulate apoptosis by targeting the apoptosis-inducing factor (AIF). Our data also revealed that AIF translocated to the nucleus to induce DNA fragmentation, and could competitively bind to HSP70 to disintegrate the HSP70-Bax (Bcl-2-associated X protein) complex, thereby activating the mitochondria apoptosis pathway by freeing Bax. The present finding therefore provides a novel mechanism that underlies the crosstalk between exosomal miRNAs and apoptosis pathway in innate immune response in invertebrates.
Assuntos
Apoptose/genética , Braquiúros/genética , Exossomos/genética , Animais , Fator de Indução de Apoptose/genética , Fator de Indução de Apoptose/metabolismo , Braquiúros/metabolismo , Braquiúros/virologia , Decápodes/genética , Decápodes/metabolismo , Decápodes/virologia , Exossomos/metabolismo , Hemócitos/imunologia , Hemócitos/metabolismo , Imunidade Inata , Infecções , MicroRNAs/genética , MicroRNAs/metabolismo , Mitocôndrias , Replicação Viral/genética , Vírus da Síndrome da Mancha Branca 1/metabolismo , Vírus da Síndrome da Mancha Branca 1/patogenicidadeRESUMO
OBJECTIVES: To assess whether dual-energy computed tomography (DECT), using conventional computed tomography or magnetic resonance imaging as a reference standard, is sufficiently accurate to differentiate intracerebral hemorrhage from contrast extravasation after endovascular thrombectomy for acute ischemic stroke. METHODS: On January 20, 2021, we searched the PubMed Medline, Embase, Web of Science, and Cochrane Library databases. QUADAS-2 was used to assess the risk of bias and applicability. Meta-analyses were performed using a bivariate random-effects model. To explore sources of heterogeneity, meta-regression analyses were performed. Deeks' funnel plot asymmetry test was used to assess publication bias. RESULTS: A total of 7 studies (269 patients, 269 focal areas) were included. The pooled mean sensitivity, specificity, and accuracy of DECT in identifying intracerebral hemorrhage from contrast extravasation after mechanical thrombectomy for acute ischemic stroke were 0.77 (95% confidence interval (CI) 0.29 to 0.96), 1 (95% CI 0.86 to 1), and 0.99 (95% CI 0.98 to 1), respectively. This evidence was of moderate certainty due to the risk of bias. Higgin's I-squared for study heterogeneity was observed for the pooled sensitivity (I2 = 78.88%) and pooled specificity (I2 = 82.12%). Moreover, Deeks' funnel plot asymmetry test revealed no publication bias (p = 0.38). CONCLUSION: DECT shows excellent accuracy and specificity in differentiating intracerebral hemorrhage from contrast extravasation after endovascular thrombectomy for acute ischemic stroke. Nevertheless, there was substantial and moderate heterogeneity among the studies. Future large-scale, prospective cohort studies are warranted to validate our findings. KEY POINTS: ⢠Dual-energy computed tomography shows excellent accuracy and specificity in differentiating intracerebral hemorrhage from contrast extravasation after endovascular thrombectomy for acute ischemic stroke. ⢠Via meta-regression analysis, we found various possible covariates, including the publication date, image analysis, index test time, time of follow-up imaging, and reference standard judgment, that had an important effect on the heterogeneity. ⢠There were no concerns regarding applicability in any of the included studies.