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1.
Sci Rep ; 12(1): 6025, 2022 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-35410352

RESUMO

Echinoderms constitute an animal phylum characterized by the pentaradial body plan. During the development from bilateral larvae to pentaradial adults, the formation of the multiple of five hydrocoel lobes, i.e., the buddings from the mesodermal coelom, is the firstly emerging pentameral character. The developmental mechanism underlying the hydrocoel-lobe formation should be revealed to understand the evolutionary process of this unique and highly derived body plan of echinoderms, although the morphogenetic mechanisms of hydrocoel lobes are largely uninvestigated. In this study, using the sea cucumber Apostichopus japonicus, in which hydrocoel is easily observable, the developmental process of hydrocoel lobes was described in detail, focusing on cell proliferation and rearrangement. Cell proliferation was not specifically distributed in the growing tips of the hydrocoel lobes, and inhibition of cell proliferation did not affect lobe formation. During lobe formation, the epithelium of the hydrocoel lobes was firstly thickened and then transformed into a simple epithelium, suggesting that tissue expansion via tissue remodeling contributes to the hydrocoel-lobe formation.


Assuntos
Pepinos-do-Mar , Stichopus , Animais , Larva
2.
Virulence ; 13(1): 458-470, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-35259068

RESUMO

Vibrio splendidus is the main opportunistic pathogen that causes skin ulcer syndrome in Apostichopus japonicus. hppDIn the present study, mutant V. splendidus with an in-frame deletion of hppDV.s. (MTVs) was constructed. The median lethal doses of wild-type V. splendidus (WTVs) and MTVs were 5.129 × 106 and 2.606 × 1010 CFU mL-1, respectively. RNA-Seq was performed using WTVs and MTVs cells at different growth stages to explore the mechanisms of the pathogenesis mediated by hppDV.s. Gene Ontology analysis showed that the expression levels of 105 genes involved in amino acid metabolism and protein binding were remarkably different between MTVs and WTVs. Kyoto Encyclopedia of Genes and Genomes analysis showed that the pathways of glutamate metabolism and flagellum assembly involved in biofilm formation and swarming motility were suppressed in MTVs. Correspondingly, the swarming motility, biofilm formation and colonisation of MTVs were remarkably decreased compared with those of WTVs. The results showed that 4-hppD catalyses tyrosine into fumarate, which could enhance glutamate metabolism and ATP production; promote flagellum assembly through the TCA cycle and lead to higher swarming, biofilm formation and colonisation abilities, to contribute to the pathogenesis of V. splendidus.


Assuntos
Stichopus , Animais , Flagelos/genética , Ácido Glutâmico , Imunidade Inata/genética , Vibrio , Virulência
3.
Food Funct ; 13(7): 3865-3878, 2022 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-35274663

RESUMO

Hyperuricemia (HUA) is the second most common metabolic disease nowadays, and is characterized by permanently increased concentrations of serum uric acid. In this study, two novel hexapeptides (GPAGPR and GPSGRP) were identified from Apostichopus japonicus hydrolysate and predicted to have xanthine oxidase (XOD) inhibitory activity by molecular docking. Their in vitro XOD inhibition rates reached 37.3% and 48.6%, respectively, at a concentration of 40 mg mL-1. Subsequently, in vivo experiments were carried out in a HUA mouse model, and we found that both peptides reduced the serum uric acid by inhibiting uric acid biosynthesis and reabsorption, as well as alleviated renal inflammation via suppressing the activation of the NLRP3 inflammasome. 16S rDNA sequencing indicated that both peptide treatments reduced the richness and diversity of the gut microbiota, altered the composition in the phylum and genus levels, but different change trends were observed in the phylum Verrucomicrobia and genera Akkermansia, Dubosiella, Alloprevotella, Clostridium unclassified and Alistipes. In addition, changes in the renal microRNA (miRNA) profiles induced by GPSGRP treatment were analyzed; 21 differentially expressed (DE) miRNAs were identified among groups, and KEGG pathway analysis indicated that their potential target genes were involved in pluripotency of stem cell regulation, mTOR signaling pathway and proteoglycans. Moreover, ten miRNAs involved in the HUA onset and alleviation were identified, which showed a high correlation with genera related to the metabolism of short-chain fatty acids, bile acids and tryptophan. This study delineated two hexapeptides as potential microbiota modulators and miRNA regulators that can ameliorate HUA.


Assuntos
Microbioma Gastrointestinal , Hiperuricemia , MicroRNAs , Stichopus , Animais , Camundongos , MicroRNAs/genética , Simulação de Acoplamento Molecular , Stichopus/metabolismo , Ácido Úrico , Xantina Oxidase
4.
Zool Res ; 43(2): 285-300, 2022 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-35238186

RESUMO

Organisms produce high levels of reactive oxygen species (ROS) to kill pathogens or act as signaling molecules to induce immune responses; however, excessive ROS can result in cell death. To maintain ROS balance and cell survival, mitophagy selectively eliminates damaged mitochondria via mitophagy receptors in vertebrates. In marine invertebrates, however, mitophagy and its functions remain largely unknown. In the current study, Vibrio splendidus infection damaged mitochondrial morphology in coelomocytes and reduced mitochondrial membrane potential (ΔΨm) and mitophagosome formation. The colocalization of mitochondria and lysosomes further confirmed that lipopolysaccharide (LPS) treatment increased mitophagy flux. To explore the regulatory mechanism of mitophagy, we cloned Bcl2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3), a common mitophagy receptor, from sea cucumber Apostichopus japonicus (AjBNIP3) and confirmed that AjBNIP3 was significantly induced and accumulated in mitochondria after V. splendidus infection and LPS exposure. At the mitochondrial membrane, AjBNIP3 interacts with microtubule-associated protein 1 light chain 3 (LC3) on phagophore membranes to mediate mitophagy. After AjBNIP3 interference, mitophagy flux decreased significantly. Furthermore, AjBNIP3-mediated mitophagy was activated by ROS following the addition of exogenous hydrogen peroxide (H2O2), ROS scavengers, and ROS inhibitors. Finally, inhibition of BNIP3-mediated mitophagy by AjBNIP3 small interfering RNA (siRNA) or high concentrations of lactate increased apoptosis and decreased coelomocyte survival. These findings highlight the essential role of AjBNIP3 in damaged mitochondrial degradation during mitophagy. This mitophagy activity is required for coelomocyte survival in A. japonicus against V. splendidus infection.


Assuntos
Stichopus , Animais , Peróxido de Hidrogênio , Mitofagia/genética , Espécies Reativas de Oxigênio/metabolismo , Stichopus/genética , Stichopus/metabolismo , Vibrio
5.
Int J Biol Macromol ; 203: 572-582, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35120936

RESUMO

RhoA (Ras homolog A) protein is a representative member of the Rho GTPase family and is involved in various cellular processes. The function of RhoA in sea cucumbers is unclear. In this study, we hypothesized that RhoA may regulate the innate immune response of Apostichopus japonicus. Our data showed that 1) the complete sequence of RhoA from A. japonicus (named AjRhoA) was 968 bp, with a high level sequence conservation across the echinoderms and other phyla; 2) tissue expression analysis showed that AjRhoA transcripts and protein exhibited higher abundance in coelomocytes, whereas the relative expression of miR-2012-5p was lower in coelomocytes; 3) interactive binding sites and a negative regulatory targeting relationship between AjRhoA and miR-2012-5p were confirmed through a dual-luciferase reporter assay and functional validation in vivo; 4) the relative expression levels of AjRhoA transcripts and protein were upregulated in coelomocytes 4- and 72-hour post infection (hpi) with Vibrio splendidus, whereas miR-2012-5p was expressed in the opposite pattern; 5) both AjRhoA silencing and miR-2012-5p overexpression suppressed the phagocytic capacity of A. japonicus compared with the control at 4 and 72 hpi. Our observations suggest that AjRhoA can regulate the pathogen-induced immune response of A. japonicus through the "AjRhoA-miR-2012-5p" module during the early infection, while miR-2012-5p plays a direct immunomodulatory role as the infection progresses.


Assuntos
MicroRNAs , Pepinos-do-Mar , Stichopus , Vibrio , Animais , Regulação da Expressão Gênica , Imunidade Inata/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Pepinos-do-Mar/genética , Stichopus/metabolismo , Vibrio/genética
6.
Mar Biotechnol (NY) ; 24(1): 151-162, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35122573

RESUMO

Polyploid breeding is widely used in aquaculture as an important area of new research. We have previously grown Apostichopus japonicus triploids with a growth advantage. The body length, body weight, and aestivation time of triploid and diploid A. japonicus were measured in this study, and the transcriptome and metabolome were used to examine the growth advantage of triploids A. japonicus. The results showed that the proportion of triploid A. japonicus with a body length of 6-12 cm and 12-18 cm was significantly higher than that of diploid A. japonicus, and triploid A. japonicus had a shorter aestivation time (39 days) than diploid (63 days). We discovered 3296 differentially expressed genes (DEGs); 13 DEGs (for example, cyclin-dependent kinase 2) related to growth advantage, immune regulation, and energy storage were screened as potential candidates. According to Gene Ontology (GO) enrichment analysis, DEGs were significantly enriched in the cytoplasm (cellular component), ATP binding process (molecular function), oxidation-reduction process (biological process), and other pathways. According to the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment data, DEGs were significantly enriched in ribosome production and other areas. We discovered 414 significant differential metabolites (SDMs), with 11 important SDMs (for example, nocodazole) linked to a growth advantage. SDMs are significantly enriched in metabolic pathways, as well as other pathways, according to the KEGG enrichment results. According to a combined transcriptome and metabolome analysis, 6 DEGs have regulatory relationships with 11 SDMs, which act on 11 metabolic pathways together. Our results further enrich the biological data of triploid A. japonicus and provide useful resources for genetic improvement of this species.


Assuntos
Fenômenos Biológicos , Stichopus , Animais , Perfilação da Expressão Gênica , Crescimento e Desenvolvimento , Stichopus/genética , Transcriptoma , Triploidia
7.
Food Funct ; 13(4): 2306-2322, 2022 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-35142318

RESUMO

Enzymatic hydrolysis-offline and membrane separation (EH-offline MS), enzymatic membrane reactor (EMR) (various operational modes), and conjoined nanofilter-purification (desalination) were used to produce highly stable antihypertensive and antioxidative peptides from ultrasonic-slurry viscosity reduced sea cucumber (A. japonicus) protein. The adoption of the optimum batch parameters by EMR-gradient diafiltration feeding (GDF), water feeding, and substrate feeding ensured a significant (p < 0.05) enhancement in protein conversion degree (PCD) by 60.39, 46.69, and 23.33%, respectively, over the conventional EH-offline MS. Also, the antihypertensive activity (ACE-inhibitory potency) of the peptides produced was in the order EMR-GDF > substrate feeding > water feeding > batch process > EH-offline MS. The EMR-GDF and nanofilter-purification produced highly digestible peptides with ACE-inhibition activities of 79.44% and 77.57% for gastric and gastrointestinal digests, respectively. Peptides with molecular weights of 1000-500 Da and 500 Da significantly contributed to the antihypertensive potency of desalinated peptides. In vitro simulated peptides showed a significant increase in the hydroxyl radical scavenging activity for gastric (77.27%) and gastrointestinal (85.32%) digests. The antioxidative stability of the produced peptides was least affected by high-temperature storage. The high arginine (Arg) and hydrophobic amino acid (HAA) content of the peptides resulted in their improved digestibility. Therefore, conjoined EMR-GDF and nanofilter-purification in the production of highly stable desalinated bioactive peptides for industrial applications could be a viable alternative.


Assuntos
Anti-Hipertensivos/farmacologia , Antioxidantes/farmacologia , Stichopus , Animais , Anti-Hipertensivos/química , Antioxidantes/química , Compostos de Bifenilo , Ensaios Enzimáticos , Filtração , Picratos
8.
Fish Shellfish Immunol ; 122: 170-180, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35150828

RESUMO

Glutaminolysis has been proved to play an irreplaceable role in vertebrate immunity, including effects on cytokine production, bacterial killing, and redox homeostasis maintenance. Our previous metabolomics analysis indicated that glutaminolysis metabolic substrates glutamine (Gln) and metabolites glutamate (Glu) were significantly lower in Skin ulceration syndrome (SUS)-diseased Apostichopus japonicus. To further delineate the role of glutaminolysis, we assayed the levels of Gln and Glu. We found that their contents in coelomocytes were decreased, accompanied by an increase in glutathione (GSH) in pathogen-challenged Apostichopus japonicus. Consistently, the mRNA transcripts of three key genes in glutaminolysis (AjASCT2, AjGOT, and AjGCS) were significantly induced. Moreover, the increased MDA and NADPH/NADP + levels in response to pathogen infection indicated that oxidative stress occurs during the immune response. The metabolic regulator AMPKß could regulate glutaminolysis in vertebrates by inducing cells to take up extracellular Gln. To explore the underlying regulatory mechanism behind glutaminolysis that occurred in coelomocytes, the full-length cDNA of AMPKß was identified from A. japonicus (designated as AjAMPKß). AjAMPKß expression was significantly induced in the coelomocytes after pathogen challenge, which was consistent with the expression of key genes of glutaminolysis. A functional assay indicated that AjAMPKß silencing by siRNA transfection could increase the levels of Gln and Glu and depress the production of GSH. Moreover, the expression of glutaminolysis-related genes was significantly inhibited, and the reduction of redox homeostasis indexes (MDA and NADPH/NADP+) was also observed. Contrastingly, AjAMPKß overexpression promoted redox homeostasis balance. Intracellular ROS is mostly responsible for breaking redox homeostasis and leading to oxidative stress, contributing to cell fate changes in immune cells. Exogenous Gln and GSH treatments could significantly reduce ROS level while the AjAMPKß silencing induced the level of ROS and accelerated the necrosis rate. All these results collectively revealed that AjAMPKß could modulate cellular redox homeostasis by affecting the glutaminolysis in A. japonicus.


Assuntos
Stichopus , Vibrio , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Homeostase , Imunidade Inata/genética , Oxirredução , Vibrio/fisiologia
9.
Antonie Van Leeuwenhoek ; 115(4): 497-503, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35149918

RESUMO

A Gram-stain negative, aerobic, rod-shaped bacterium, designated 126T, was isolated from the intestinal content of a sea cucumber, Apostichopus japonicus, in China. Strain 126T was found to grow optimally at 25-28 °C and pH 7.5-8.0 in marine 2216 E medium, with tolerance of 1-7% (w/v) NaCl. Strain 126T is motile by means of one to several polar flagella. The dominant fatty acids of strain 126T were identified as C16:1 ω7c/C16:1 ω6c (29.5%), C18:1 ω7c/C18:1 ω6c (19.8%) and C16:0 (16.7%). The respiratory quinone was found to be Q-8. The polar lipid profile was found to be mainly composed of phosphatidylglycerol and phosphatidylethanolamine. The total length of the draft genome is approximately 4.2 × 106 bp, encoding 3655 genes and 3576 coding sequences. The G + C content of the genomic DNA is 48.0%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 126T belongs to the genus Neiella and is closely related to Neiella marina J221T (96.5%). Genomic comparisons of 126T to N. marina J221T revealed that they had similar genome size, G + C content and complement of clusters of orthologous groups. However, average nucleotide identity and digital DNA-DNA hybridization values between strains126T and N. marina J221T was 75.5% and 19.7%, which could distinguish the strains. On the basis of these phenotypic and genotypic data, strain 126T is concluded to represent a novel species, for which the name Neiella holothuriorum sp. nov. is proposed. The type strain is 126T (= GDMCC 1.2530T = KCTC 82829T).


Assuntos
Pepinos-do-Mar , Stichopus , Animais , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Ácidos Graxos/análise , Hibridização de Ácido Nucleico , Fosfolipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Stichopus/genética , Stichopus/microbiologia
10.
Fish Shellfish Immunol ; 121: 135-141, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34998985

RESUMO

Matrix metalloproteinases (MMPs) are an important family of proteinases involved in various physiological processes and associated with the immune response. However, the role of MMPs in the immune response remains unclear. To explore the possible role of MMPs in innate immunity, this study selected the MMP-16 gene encoding peptidoglycan (PGN) binding domain identified in the sea cucumber Apostichopus japonicus (named AjMMP-16, GenBank accession No. AQT26486) for microbial polysaccharide-induced transcriptional expression analysis by quantitative real-time PCR, correlation analysis with nine representative genes from A. japonicus immune pathways in microbial polysaccharide-induced transcriptional expression by using Pearson's correlation test, and prokaryotic recombinant expression. Next, its recombinant protein was employed for microbial polysaccharide-binding analysis with ELISA and bacterial binding analysis with the indirect immunofluorescence method. The results showed that AjMMP-16 was significantly induced by diaminopimelic acid (DAP)-type PGN, lipopolysaccharide, mannan, and ß-1,3-glucan and was closely correlated with myeloid differentiation factor 88 (MyD88) in microbial polysaccharide-induced transcriptional expression. In addition, recombinant AjMMP-16 bound to lysine-type PGN, DAP-type PGN, lipopolysaccharide, mannan, ß-1,3-glucan, Vibrio splendidus, Pseudoalteromonas nigrifaciens, Shewanella baltica, Bacillus cereus, Escherichia coli, and Staphylococcus aureus. These results suggest that AjMMP-16 might act as a pattern recognition receptor in innate immunity and play an important role in initiating the MyD88-dependent Toll-like receptor signaling pathway.


Assuntos
Imunidade Inata , Metaloproteinase 16 da Matriz , Receptores de Reconhecimento de Padrão , Stichopus , Animais , Bactérias , Lipopolissacarídeos , Mananas , Metaloproteinase 16 da Matriz/imunologia , Fator 88 de Diferenciação Mieloide , Stichopus/genética , Stichopus/imunologia
11.
FEBS Open Bio ; 12(3): 664-674, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35090095

RESUMO

Ferritin is considered to be an ubiquitous and conserved iron-binding protein that plays a crucial role in iron storage, detoxification, and immune response. Although ferritin is of critical importance for almost all kingdoms of life, there is a lack of knowledge about its role in the marine invertebrate sea cucumber (Apostichopus japonicus). In this study, we characterized the first crystal structure of A. japonicus ferritin (AjFER) at 2.75 Å resolution. The structure of AjFER shows a 4-3-2 symmetry cage-like hollow shell composed of 24 subunits, mostly similar to the structural characteristics of other known ferritin species, including the conserved ferroxidase center and 3-fold channel. The 3-fold channel consisting of three 3-fold negative amino acid rings suggests a potential pathway in which metal ions can be first captured by Asp120 from the outside environment, attracted by His116 and Cys128 when entering the channel, and then transferred by Glu138 from the 3-fold channel to the ferroxidase site. Overall, the presented crystal structure of AjFER may provide insights into the potential mechanism of the metal transport pathway for related marine invertebrate ferritins.


Assuntos
Pepinos-do-Mar , Stichopus , Animais , Cristalografia , Ferritinas/química , Ferritinas/metabolismo , Invertebrados/metabolismo , Pepinos-do-Mar/metabolismo , Stichopus/metabolismo
12.
Food Res Int ; 151: 110841, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34980380

RESUMO

Inflammatory bowel disease afflicted individuals and most medications have adverse effects. The objection of this study is to investigate whether the major yolk protein (MYP) could aid in the remission of colitis. The function of MYP on acute colitis was assessed through a dextran sulfate sodium -induced colitis mice model. Compared to the model group, the anti-inflammatory cytokines increased significantly in the MYP group, whereas the pro-inflammatory cytokines were not significantly different between the model and treatment group. The results also showed that supplementation of MYP improved the shift in microbial community composition of mice with colitis induced by DSS. In addition, MYP supplementation enriched the contents of fecal short-chain fatty acids. The alleviation of MYP on the colitis was probably related to repair the dysbiosis state of colonic microbiota, which thus induced an increase in short-chain fatty acids level and secrete anti-inflammatory cytokines (IL-4 and IL-10). In sum, oral MYP may be a potential candidate for the attenuating of acute colitis.


Assuntos
Colite , Microbioma Gastrointestinal , Pepinos-do-Mar , Stichopus , Animais , Colite/induzido quimicamente , Colite/tratamento farmacológico , Sulfato de Dextrana , Disbiose/tratamento farmacológico , Proteínas do Ovo , Camundongos , Camundongos Endogâmicos C57BL
13.
Sci Total Environ ; 820: 153168, 2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35051475

RESUMO

Heat stress poses an increasing threat for the marine invertebrate Apostichopus japonicus. Histone lysine acetylation is a central chromatin modification for epigenetic regulation of gene expression during stress response. In this study, a genome-wide characterization for acetylated lysine 9 on histone H3 (H3K9ac) binding regions in normal temperature (18 °C) and heat-stress conditions (26 °C) via ChIP-seq were carried out. The results that revealed H3K9ac was an extensive epigenetic modulation in A. japonicus. The GO terms "regulation of transcription, DNA-templated" and "transcription coactivator activity" were significantly enriched in both groups. Particularly, various transcriptional factors (TFs) families showed notable modification of H3K9ac. Differentially acetylated regions (DARs) with H3K9ac modification under heat stress were identified with 24 hyperacetylated and 23 hypoacetylated peaks, respectively. We further examined the transcriptional expression for 13 genes with dysregulated H3K9ac level in the promoter regions by qRT-PCR. Combined H3K9ac ChIP-seq characteristics with the transcriptional expression, 5 up-up genes (ZCCHC3, RPA70, MTRR, ß-Gal and PHTF2) and 2 down-down genes (PRPF39 and BSL78_10147) were identified. Surprisingly, the increasing mRNA expression of NECAP1 under heat stress was negatively related to the decreasing H3K9ac level in its promoter region. Our research is the first genome-wide characterization for the epigenetic modification H3K9ac in A. japonicus, and will help to advance the understanding of the roles of H3K9ac in transcriptional regulation under heat-stress condition.


Assuntos
Pepinos-do-Mar , Stichopus , Acetilação , Animais , Sequenciamento de Cromatina por Imunoprecipitação , Epigênese Genética , Resposta ao Choque Térmico , Código das Histonas , Humanos , Pepinos-do-Mar/genética , Stichopus/genética
14.
Artigo em Inglês | MEDLINE | ID: mdl-34715349

RESUMO

Sea cucumbers Apostichopus japonicus (3.54 ± 0.01 g of wet weight) were exposed to five concentrations of dietary hexavalent chromium (Cr6+) [0 (control), 100, 200, 400, and 800 mg Cr6+/kg dry weight] amended with K2Cr2O7 for 30 days. The bioaccumulation and immune responses [antioxidant enzymes: superoxide dismutase (SOD) and catalase (CAT); hydrolytic enzymes: acid phosphatase (ACP) and alkaline phosphatase (AKP)] of sea cucumbers were subsequently evaluated. This study found that the order of Cr accumulation in the experimental tissues was respiratory tree > intestine > body wall. Significantly lower SOD activities occurred in the 400 mg/kg group compared to that in the control group. Higher dietary Cr6+ exposure (400 and 800 mg Cr6+/kg dry weight) did not negatively alter the CAT activities, but significantly inhibited CAT activities in 100 mg/kg group, compared to control group. ACP activities in groups 200, 400 and 800 mg/kg were significantly lower than those in control group, while no significant differences occurred in AKP activities among groups. The present study provides important information into the bioaccumulation and immune responses of the sea cucumber A. japonicus in response to chronic dietary Cr6+ exposure.


Assuntos
Cromo/toxicidade , Poluentes Ambientais/toxicidade , Stichopus/efeitos dos fármacos , Stichopus/metabolismo , Animais , Antioxidantes/metabolismo , Esquema de Medicação , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Stichopus/imunologia , Distribuição Tecidual
15.
Dev Comp Immunol ; 127: 104298, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34662683

RESUMO

Tissue inhibitors of metalloproteinases (TIMPs) serve as matrix metalloproteinase (MMP) inhibitors in the pathogenesis of inflammatory diseases in vertebrates. We cloned and characterised the TIMP1 gene from Apostichopus japonicus using RACE approaches (designated as AjTIMP1). For Vibrio splendidus-challenged sea cucumbers, the peak expression of AjTIMP1 mRNAs in coelomocytes was detected at 24 h (23.44-fold) and remained at high levels (4.01-fold) until 72 h. Similarly, AjTIMP1 expression was upregulated in primary coelomocytes exposed to 10 µg mL-1 LPS. AjTIMP1 was expressed in all tissues, and the highest expression was observed in the body wall. Functional investigation revealed an imbalance in the ratio of AjMMP1/AjTIMP1 in the skin ulceration syndrome (SUS) diseased group; it was sharply up-regulated to 3.97:1 compared with the healthy group. Furthermore, when AjTIMP1 was knocked down using small interfering RNA (siRNA-KD) to 0.4-fold, AjMMP1 and AjMMP19 were upregulated to 1.99- and 1.85-fold, respectively. AjTIMP1 siRNA-KD can promote ROS production by 26.2%, whereas AjMMP1 siRNA-KD can eliminate the increase in ROS. In inflamed tissues, collagen I and III levels were decreased by 33.1% and 33.6%, respectively, in the AjTIMP1 siRNA group at 24 h AjTIMP1 was involved in the inflammatory response by mediating ROS formation and collagen degradation.


Assuntos
Pepinos-do-Mar , Stichopus , Vibrio , Animais , Imunidade Inata/genética , Inflamação , Metaloproteinase 1 da Matriz/genética , Espécies Reativas de Oxigênio/metabolismo , Pepinos-do-Mar/genética , Pepinos-do-Mar/metabolismo , Stichopus/genética , Stichopus/metabolismo , Vibrio/fisiologia
16.
J Proteomics ; 251: 104396, 2022 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-34673268

RESUMO

The sea cucumber Apostichopus japonicus is an important aquaculture species in China because of its high nutritional and medicinal values. Gender, as a factor affecting the physiology of organisms, is always considered when improving the breeding efficiency of economically important animals. In the present study, protein expression profiles of the gonads and tube feet of male and female A. japonicus were investigated using a comparative proteomics approach. A set of 7499 proteins were identified, which covered a broad range of functions based on function annotations. A significant difference in protein expression profiles was observed between the gonads and tube feet of A. japonicus; gonads showed more apparent gender differences than tube feet. Moreover, the findings revealed that male A. japonicus had more specific functions and most of these functions were associated with energy consumption. Further analyses suggested that the regulation of ERK activity and the capacity of tyrosine production and virus immunity might be more powerful in male and female A. japonicus, respectively. Some candidate proteins were also recognized as potential targets for gender identification of A. japonicus. Overall, our study provides new insights into the understanding of molecular mechanisms underlying gender-based physiological differences in A. japonicus. SIGNIFICANCE: The current study aimed to reveal gender differences in the physiological characteristics of gonads and tube feet of the sea cucumber A. japonicus. To the best of our knowledge, this is the first proteomics study to analyze the differences in the protein expression profiles of external organs between male and female A. japonicus. The analysis revealed gender differences in the protein expression profiles of both gonads and tube feet of A. japonicus, and the gender differences in gonads were quite apparent. Moreover, according to the recognition of differentially expressed proteins and the enrichment analyses based on Kyoto Encyclopedia of Genes and Genomes, a draft view of how the physiological functions of A. japonicus were affected by gender was obtained. Male A. japonicus could have more specific functions related to energy consumption than females. The regulation of ERK activity and virus immunity might be more robust in male and female A. japonicus, respectively. Some candidate proteins were also recognized as potential targets for gender identification of A. japonicus. The findings presented here will improve the understanding of researchers about the molecular mechanisms underlying gender-based differences in A. japonicus and contribute to the meticulous breeding of A. japonicus.


Assuntos
Pepinos-do-Mar , Stichopus , Animais , Feminino , Gônadas , Masculino , Proteômica , Pepinos-do-Mar/metabolismo , Fatores Sexuais , Stichopus/genética
17.
J Immunol ; 208(2): 464-479, 2022 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-34965964

RESUMO

Inflammation participates in host defenses against infectious agents and contributes to the pathophysiology of many diseases. IL-17 is a well-known proinflammatory cytokine that contributes to various aspects of inflammation in vertebrates. However, the functional role of invertebrate IL-17 in inflammatory regulation is not well understood. In this study, we first established an inflammatory model in the Vibrio splendidus-challenged sea cucumber Apostichopus japonicus (Echinodermata). Typical inflammatory symptoms, such as increased coelomocyte infiltration, tissue vacuoles, and tissue fractures, were observed in the V. splendidus-infected and diseased tissue of the body wall. Interestingly, A. japonicus IL-17 (AjIL-17) expression in the body wall and coelomocytes was positively correlated with the development of inflammation. The administration of purified recombinant AjIL-17 protein also directly promoted inflammation in A. japonicus Through genome searches and ZDOCK prediction, a novel IL-17R counterpart containing FNIII and hypothetical TIR domains was identified in the sea cucumber genome. Coimmunoprecipitation, far-Western blotting, and laser confocal microscopy confirmed that AjIL-17R could bind AjIL-17. A subsequent cross-linking assay revealed that the AjIL-17 dimer mediates the inflammatory response by the specific binding of dimeric AjIL-17R upon pathogen infection. Moreover, silencing AjIL-17R significantly attenuated the LPS- or exogenous AjIL-17-mediated inflammatory response. Functional analysis revealed that AjIL-17/AjIL-17R modulated inflammatory responses by promoting A. japonicus TRAF6 ubiquitination and p65 nuclear translocation and evenly mediated coelomocyte proliferation and migration. Taken together, our results provide functional evidence that IL-17 is a conserved cytokine in invertebrates and vertebrates associated with inflammatory regulation via the IL-17-IL-17R-TRAF6 axis.


Assuntos
Citocinas/imunologia , Interleucina-17/metabolismo , Receptores de Interleucina-17/metabolismo , Stichopus/imunologia , Vibrio/imunologia , Animais , Proliferação de Células/fisiologia , Genoma/genética , Inflamação/imunologia , Interleucina-17/genética , Interferência de RNA , RNA Interferente Pequeno/genética , Receptores de Interleucina-17/genética , Stichopus/genética , Stichopus/microbiologia , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismo , Fator de Transcrição RelA/metabolismo , Ubiquitinação
18.
Food Chem ; 368: 130768, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34392120

RESUMO

Apoptosis plays a critical role in sea cucumber autolysis. To investigate the ultraviolet (UV)-induced apoptosis, sea cucumbers with and without injection of BAPTA-AM (cytosolic calcium chelator) were exposed to UV (15 W/m2) for 30 min. The results showed that UV irradiation caused several changes in sea cucumber coelomocytes, including calcium imbalance, abnormal morphology of endoplasmic reticulum, upregulation of pro-apoptotic proteins CRT, CHOP, and caspases 9 and 3, and downregulation of anti-apoptotic protein Bcl-2. A comparison between the two groups showed that injection of the calcium chelator into sea cucumbers helped maintain coelomocyte intracellular calcium homeostasis and suppressed other abnormal changes caused by ER stress, indicating apoptosis in sea cucumbers is mediated by calcium imbalance and follows the activation of the ER stress pathway. Therefore, this study broadens understanding of the apoptotic mechanism involved in sea cucumber autolysis, which is helpful in developing preservative agents for sea cucumbers.


Assuntos
Pepinos-do-Mar , Stichopus , Animais , Autólise , Cálcio , Homeostase
19.
Food Chem ; 373(Pt A): 131368, 2022 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-34717088

RESUMO

Thermal processing is the most frequently adopted processing technology for sea cucumbers, which can significantly affect their protein composition. In this paper, three thermal processing methods high pressure steaming (HPS), atmospheric pressure boiling (APB), and atmospheric pressure steaming (APS) were adopted and protein compositions of both body walls and cooking liquors by thermal processing stichopus japonicus were systematically analysis by proteomic strategy. The total proteins loss rates of body walls were 11.6%, 13.0%, and 14.8% for HPS, APS, and APB methods, respectively. However, the main types of protein composition were retained. Similar mechanisms of protein loss may exist even if different thermal processing were applied. The most frequent hydrolysis sites in thermal processing were phenylalanine, leucine, asparagine, and tyrosine at both C and N terminals. This study provides theoretical guidance for optimizing the industry thermal processing of sea cucumbers.


Assuntos
Pepinos-do-Mar , Stichopus , Animais , Cromatografia Líquida de Alta Pressão , Proteômica , Espectrometria de Massas em Tandem
20.
Chemosphere ; 287(Pt 1): 132062, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34526273

RESUMO

Microplastics (MPs; ≤ 5 mm) have become a potential threat to human health due to the widespread detection of MPs in foods consumed by humans. Here, we investigated the potential of MP occurrence in the main edible part of the most valuable species of sea cucumbers (Apostichopus japonicus). Laboratory experiments showed that fluorescent MPs and microfibers (MFs) could transfer into the body wall of the sea cucumber. The evidence revealed that these MPs enter the body wall via the outer surface. Although these MPs decreased after the sea cucumbers were transferred to clean water, traces of MPs (at least one MP particle) were found up to 60 d post-transfer. To validate these laboratory observations, sea cucumber samples were collected from the field. MPs were found in 86% of live and processed sea cucumber samples. The MP abundances in the field samples ranged from 0-15 MPs animal-1 and 0-2 MP g-1. The isolated MPs were mainly MFs, constituting 81% of MPs, followed by fragments, films, and beads. Fourier transform infrared spectroscopy revealed that the polymer composition of the isolated MPs mainly included rayon, followed by polyester and chlorinated polyethylene. The findings of this study demonstrated that the body walls of farmed and processed sea cucumbers contain MPs, thus highlighting the need to control MP pollution during the farming and processing of sea cucumbers.


Assuntos
Pepinos-do-Mar , Stichopus , Poluentes Químicos da Água , Animais , Humanos , Microplásticos , Plásticos , Poluentes Químicos da Água/análise
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