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Gene Expression Patterns : GEP Dec 2023Peroxidase genes (Prdx) encode a family of antioxidant proteins, which can protect cells from oxidative damage by reducing various cellular peroxides. This study...
Peroxidase genes (Prdx) encode a family of antioxidant proteins, which can protect cells from oxidative damage by reducing various cellular peroxides. This study investigated the spatiotemporal expression patterns of gene members in this family during the early development of Xenopus tropicalis. Real-time quantitative PCR showed that all members of this gene family have a distinct temporal expression pattern during the early development of X. tropicalis embryos. Additionally, whole mount in situ hybridization revealed that individual prdx genes display differential expression patterns, with overlapping expression in lymphatic vessels, pronephros, proximal tubule, and branchial arches. This study provides a basis for further study of the function of the prdx gene family.
Topics: Animals; Xenopus; Xenopus Proteins; Real-Time Polymerase Chain Reaction; Embryonic Development; Gene Expression Regulation, Developmental; Xenopus laevis
PubMed: 37844856
DOI: 10.1016/j.gep.2023.119345 -
Frontiers in Immunology 2021(Cymothoidea, Isopoda) is a generalist crustacean parasite that negatively affects the economic sustainability of European sea bass () aquaculture in the North-East...
(Cymothoidea, Isopoda) is a generalist crustacean parasite that negatively affects the economic sustainability of European sea bass () aquaculture in the North-East Mediterranean. While mortalities are observed in fry and fingerlings, infection in juvenile and adult fish result in approximately 20% growth delay. A transcriptomic analysis (PCR array, RNA-Seq) was performed on organs (tongue, spleen, head kidney, and liver) from infected vs. -free sea bass fingerlings. Activation of local and systemic immune responses was detected, particularly in the spleen, characterized by the upregulation of cytokines (also in the tongue), a general reshaping of the immunoglobulin (Ig) response and suppression of T-cell mediated responses. Interestingly, starvation and iron transport and metabolism genes were strongly downregulated, suggesting that the parasite feeding strategy is not likely hematophagous. The regulation of genes related to growth impairment and starvation supported the growth delay observed in infected animals. Most differentially expressed (DE) transcripts were exclusive of a specific organ; however, only in the tongue, the difference between infected and uninfected fish was significant. At the attachment/feeding site, the pathways involved in muscle contraction and intercellular junction were the most upregulated, whereas the pathways involved in fibrosis (extracellular matrix organization, collagen formation, and biosynthesis) were downregulated. These results suggest that parasite-inflicted damage is successfully mitigated by the host and characterized by regenerative processes that prevail over the reparative ones.
Topics: Animals; Bass; Cytokines; Fish Diseases; Gene Expression Profiling; Head Kidney; Isopoda; Liver; Mediterranean Sea; Parasitic Diseases, Animal
PubMed: 33777043
DOI: 10.3389/fimmu.2021.645607 -
Fish & Shellfish Immunology May 2020Astragalus polysaccharides (APS) have been widely used as immunopotentiators in aquaculture, however, the best way of their administration remains to be explored. In the...
Astragalus polysaccharides (APS) have been widely used as immunopotentiators in aquaculture, however, the best way of their administration remains to be explored. In the present study, APS liposome (APSL) was prepared by film dispersion-ultrasonic method. The optimal conditions of APSL preparation were determined by response surface methodology, with a ratio of 10:1 (w/w) for soybean lecithin to APS and 8:1 (w/w) for soybean lecithin to cholesterol, and an ultrasound time of 15 min, which produced an encapsulation efficiency of 73.88 ± 0.88% of APSL. In vivo feeding experiments in large yellow croaker showed that both APS and APSL could enhance the contents of serum total protein (TP) and albumin (ALB), activities of serum non-specific immune enzymes such as acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM), and phagocytic activity of head kidney macrophages. Meanwhile, they both increased the activities of serum antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) and reduced the content of final lipid peroxidation product malondialdehyde (MDA) in serum, thus exhibiting the antioxidant effects. In vitro experiments on primary head kidney macrophages (PKM) showed that both APS and APSL inhibited ROS production, but obviously enhanced NO production and phagocytic activity of PKM. Furthermore, expression levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α), IFN-γ, and iNOS in PKM were significantly up-regulated after APS and APSL treatments, but no expression change of IFN-h was observed. Taken together, our results showed that both APS and APSL could improve several immune parameters and antioxidant ability of large yellow croaker either in vivo or in vitro, and the efficacy of APSL was markedly better than APS. These findings therefore indicated that the immunomodulatory and antioxidant activities of APS could be enhanced after encapsulated with liposome, and APSL may represent a potential drug delivery system of APS for development of immunoenhancers in aquaculture.
Topics: Animals; Antioxidants; Aquaculture; Astragalus Plant; Blood Proteins; Head Kidney; Immunologic Factors; Liposomes; Macrophages; Perciformes; Phagocytosis; Plant Extracts; Serum Albumin
PubMed: 32142872
DOI: 10.1016/j.fsi.2020.03.004 -
BMC Genomics Nov 2022Infectious Salmon Anaemia virus (ISAV) is an orthomyxovirus responsible for large losses in Atlantic salmon (Salmo salar) aquaculture. Current available treatments and...
BACKGROUND
Infectious Salmon Anaemia virus (ISAV) is an orthomyxovirus responsible for large losses in Atlantic salmon (Salmo salar) aquaculture. Current available treatments and vaccines are not fully effective, and therefore selective breeding to produce ISAV-resistant strains of Atlantic salmon is a high priority for the industry. Genomic selection and potentially genome editing can be applied to enhance the disease resistance of aquaculture stocks, and both approaches can benefit from increased knowledge on the genomic mechanisms of resistance to ISAV. To improve our understanding of the mechanisms underlying resistance to ISAV in Atlantic salmon we performed a transcriptomic study in ISAV-infected salmon with contrasting levels of resistance to this virus.
RESULTS
Three different tissues (gills, head kidney and spleen) were collected on 12 resistant and 12 susceptible fish at three timepoints (pre-challenge, 7 and 14 days post challenge) and RNA sequenced. The transcriptomes of infected and non-infected fish and of resistant and susceptible fish were compared at each timepoint. The results show that the responses to ISAV are organ-specific; an important response to the infection was observed in the head kidney, with up-regulation of immune processes such as interferon and NLR pathways, while in gills and spleen the response was more moderate. In addition to immune related genes, our results suggest that other processes such as ubiquitination and ribosomal processing are important during early infection with ISAV. Moreover, the comparison between resistant and susceptible fish has also highlighted some interesting genes related to ubiquitination, intracellular transport and the inflammasome.
CONCLUSIONS
Atlantic salmon infection by ISAV revealed an organ-specific response, implying differential function during the infection. An immune response was observed in the head kidney in these early timepoints, while gills and spleen showed modest responses in comparison. Comparison between resistance and susceptible samples have highlighted genes of interest for further studies, for instance those related to ubiquitination or the inflammasome.
Topics: Animals; Isavirus; Head Kidney; Salmo salar; Spleen; Gills; Transcriptome; Inflammasomes
PubMed: 36443659
DOI: 10.1186/s12864-022-09007-4 -
The Science of the Total Environment Jun 2024PFAAs (Perfluoroalkyl acids) are a class of bioaccumulative, persistent and ubiquitous environmental contaminants which primarily occupy the hydrosphere and its...
PFAAs (Perfluoroalkyl acids) are a class of bioaccumulative, persistent and ubiquitous environmental contaminants which primarily occupy the hydrosphere and its sediments. Currently, a paucity of toxicological information exists for short chain PFAAs and complex mixtures. In order to address these knowledge gaps, we performed a 3-week, aqueous exposure of rainbow trout to 3 different concentrations of a PFAA mixture (50, 100 and 500 ng/L) modeled after the composition determined in Lake Ontario. We conducted an additional set of exposures to individual PFAAs (25 nM each of PFOS (12,500 ng/L), PFOA (10,300 ng/L), PFBS (7500 ng/L) or PFBA (5300 ng/L) to evaluate differences in biological response across PFAA congeners. Untargeted proteomics and phosphorylated metabolomics were conducted on the blood plasma and head kidney tissue to evaluate biological response. Plasma proteomic responses to the mixtures revealed several unexpected outcomes including Similar proteomic profiles and biological processes as the PFOS exposure regime while being orders of magnitude lower in concentration and an atypical dose response in terms of the number of significantly altered proteins (FDR < 0.1). Biological pathway analysis revealed the low mixture, medium mixture and PFOS to significantly alter (FDR < 0.05) a number of processes including those involved in lipid metabolism, oxidative stress and the nervous system. We implicate plasma increases in PPARD and PPARG as being directly related to these biological processes as they are known to be important regulators in all 3 processes. In contrast to the blood plasma, the high mixture and PFOA exposure regimes caused the greatest change to the head kidney proteome, altering many proteins being involved in lipid metabolism, oxidative stress and inflammation. Our findings support the pleiotropic effect PFAAs have on aquatic organisms at environmentally relevant doses including those on PPAR signaling, metabolic dysregulation, immunotoxicity and neurotoxicity.
Topics: Animals; Water Pollutants, Chemical; Oncorhynchus mykiss; Fluorocarbons; Proteome; Head Kidney
PubMed: 38615763
DOI: 10.1016/j.scitotenv.2024.172389 -
Developmental and Comparative Immunology Jan 2021Infections are able to trigger epigenetic modifications; however, epigenetic-mediating infections in the immune system in fish is currently unavailable. Within this...
Infections are able to trigger epigenetic modifications; however, epigenetic-mediating infections in the immune system in fish is currently unavailable. Within this purpose, zebrafish were immune-stimulated with three lipopolysaccharides (LPS) during sex differentiation. Methylation patterns of three immune genes were studied by a candidate gene approach together with gene expression analysis, and in adulthood, sex ratios were determined. It was shown that the entrance of LPS was through the gills and accumulated in the pronephros. Significant hypomethylation levels of CASP9 and a significant CpG site for IL1β after Pseudomonas aeruginosa LPS exposure were found. No methylation difference was observed for TNFα. Gene expression and correlation data differed among studied genes. Sex ratios showed a feminization in dose and LPS strain-dependent manner. Here, it is provided epigenetic regulatory mechanisms derived by innate response and the first evidence of possible epigenetic interactions between the immune and reproductive systems.
Topics: Animals; Caspase 9; CpG Islands; DNA Methylation; Epigenesis, Genetic; Gene Expression Profiling; Genetic Association Studies; Gonads; Immunity; Immunity, Innate; Immunization; Interleukin-1beta; Lipopolysaccharides; Pseudomonas aeruginosa; Reproduction; Sex Differentiation; Sex Ratio; Zebrafish; Zebrafish Proteins
PubMed: 32888969
DOI: 10.1016/j.dci.2020.103848 -
Fish & Shellfish Immunology Dec 2022CD4 and CD8 molecules play an important role in the identification of T lymphocytes, and diverse among fish species. In this study, CD4-1 and CD8α gene of spotted sea...
CD4-1 and CD8α T lymphocytes subsets in spotted sea bass (Lateolabrax maculatus) and comparison on antigenicity of T lymphocytes subsets in other three marine fish species.
CD4 and CD8 molecules play an important role in the identification of T lymphocytes, and diverse among fish species. In this study, CD4-1 and CD8α gene of spotted sea bass (Lateolabrax maculatus) were cloned, polyclonal antibodies against CD4-1 (CD4-1 pAbs) and CD8α (CD8α pAbs) were produced, respectively. And the variations in CD4-1 and CD8α T-lymphocytes in spotted sea bass and the cross-reactivity with leukocytes in pearl gentian grouper (Epinephelus fuscoguttatus x E. lanceolatus), schlegel's black rockfish (Sebastes schlegelii) and flounder (Paralichthys olivaceus) were investigated using CD4-1 pAbs and CD8α pAbs. The results showed that CD4-1 molecule ORF was 1413 bp and CD8α was 690 bp, both molecules are transmembrane glycoproteins with high amino acid homology to grouper. The CD4-1 pAbs specifically recognized both the CD4-1 recombinant and natural proteins, as does the CD8α pAbs to CD8α molecule, and no cross-reactivity between the two antibodies. CD4-1 and CD8α T lymphocytes were detected in peripheral blood, spleen and head kidney leukocytes in spotted sea bass. In cross-reactivity assay with other three fish, CD4-1 pAbs could recognize the lymphocytes from pearl gentian grouper and schlegel's black rockfish, both with highest proportions in the spleen leukocytes, 5.3 ± 0.4% and 2.6 ± 0.3%, respectively, and CD8α pAbs could only recognize the lymphocytes in pearl gentian grouper, and no cross-reactivities to lymphocytes of flounder. These data suggested that the CD4-1 and CD8α molecules varied by fish species in the genes features and antigenicity, which might result in the diversities of T lymphocytes subpopulations. This will be a key to elucidating the classification and evolution of T lymphocytes in fish.
Topics: Animals; Bass; Flounder; Head Kidney; Perciformes; Leukocytes; CD4 Antigens
PubMed: 36210001
DOI: 10.1016/j.fsi.2022.10.004 -
Kidney International Mar 2021Although a rare disease, bilateral congenital anomalies of the kidney and urinary tract (CAKUT) are the leading cause of end stage kidney disease in children....
Although a rare disease, bilateral congenital anomalies of the kidney and urinary tract (CAKUT) are the leading cause of end stage kidney disease in children. Ultrasound-based prenatal prediction of postnatal kidney survival in CAKUT pregnancies is far from accurate. To improve prediction, we conducted a prospective multicenter peptidome analysis of amniotic fluid spanning 140 evaluable fetuses with CAKUT. We identified a signature of 98 endogenous amniotic fluid peptides, mainly composed of fragments from extracellular matrix proteins and from the actin binding protein thymosin-β4. The peptide signature predicted postnatal kidney outcome with an area under the curve of 0.96 in the holdout validation set of patients with CAKUT with definite endpoint data. Additionally, this peptide signature was validated in a geographically independent sub-cohort of 12 patients (area under the curve 1.00) and displayed high specificity in non-CAKUT pregnancies (82 and 94% in 22 healthy fetuses and in 47 fetuses with congenital cytomegalovirus infection respectively). Change in amniotic fluid thymosin-β4 abundance was confirmed with ELISA. Knockout of thymosin-β4 in zebrafish altered proximal and distal tubule pronephros growth suggesting a possible role of thymosin β4 in fetal kidney development. Thus, recognition of the 98-peptide signature in amniotic fluid during diagnostic workup of prenatally detected fetuses with CAKUT can provide a long-sought evidence base for accurate management of the CAKUT disorder that is currently unavailable.
Topics: Amniotic Fluid; Animals; Child; Female; Humans; Kidney; Kidney Diseases; Peptides; Pregnancy; Prospective Studies; Urinary Tract; Urogenital Abnormalities; Zebrafish
PubMed: 32750455
DOI: 10.1016/j.kint.2020.06.043 -
Toxicology Letters Jun 2021Due to an increasing demand for testing of new and existing chemicals and legal restrictions for the use of animals, there is a strong need for alternative approaches to...
Due to an increasing demand for testing of new and existing chemicals and legal restrictions for the use of animals, there is a strong need for alternative approaches to assess systemic toxicity. Embryonic and larval zebrafish (Danio rerio) are increasingly recognized as a promising alternative whole-animal model that may be able to overcome limitations of cell-based in vitro assays and bridge the gap between high-throughput in vitro screening and low-throughput in vivo tests in animals. Despite the relatively simple anatomical structure of the zebrafish larval kidney (pronephros) - composed of only two nephrons - the pronephros shares major functions and cell types with mammalian nephrons. Glomerular filtration begins at 48 h post fertilization. The aim of the present study was to investigate if early zebrafish larvae might be a suitable model for nephrotoxicity testing. On day 3 post fertilization, larval zebrafish were treated with selected nephrotoxins (aristolochic acid, cadmium chloride, potassium bromate, ochratoxin A, gentamicin) for 48 h. Histological evaluation of zebrafish larvae exposed to model nephrotoxins revealed tubule injury as evidenced by dilated tubules with loss of the brush border, tubule cell necrosis and disorganization of the tubular epithelium. These changes were most severe after treatment with gentamicin, which also impaired pronephros function as evidenced by reduced clearance of FITC-dextran. Whole-mount in situ hybridization showing loss of cdh17 expression revealed site-specific injury to the proximal tubule segment. Analysis of genes previously identified as novel biomarkers of kidney injury in mammals showed upregulation of the kidney injury marker genes heme oxygenase 1 (hmox1), clusterin (clu), secreted phosphoprotein/osteopontin (spp1), connective tissue growth factor (ctgf) and kim-1 (havcr-1) in response to nephrotoxin treatment, although the response of individual genes varied across compounds. Consistent with the severity of lesions and impaired kidney function, the most prominent gene expression changes occurred in larvae exposed to gentamicin. Overall, our results suggest that larval zebrafish may be a suitable alternative model organism for nephrotoxicity screening, yet further improvements and integration with quantitative in vitro to in vivo extrapolation will be needed to predict human toxicity.
Topics: Animals; Biomarkers; Cadherins; Central Nervous System; Disease Models, Animal; Gene Expression Regulation, Developmental; Kidney; Larva; Toxicity Tests; Zebrafish; Zebrafish Proteins
PubMed: 33722575
DOI: 10.1016/j.toxlet.2021.03.005 -
Scientific Reports Feb 2021A wide variety of environmental contaminants has been shown to disrupt immune functions of fish and may compromise their defense capability against pathogens....
A wide variety of environmental contaminants has been shown to disrupt immune functions of fish and may compromise their defense capability against pathogens. Immunotoxic effects, however, are rarely considered in ecotoxicological testing strategies. The aim of this study was to systematically evaluate the suitability of an in vitro immuno-assay using selected fish immune parameters to screen for chemicals with known immunotoxic potential and to differentiate them from non-immunotoxicants. Non-stimulated and lipopolysaccharide-stimulated head kidney leukocytes of rainbow trout (Oncorhynchus mykiss) were exposed for 3 h or 19 h to chemicals with different modes of action. As immune parameters, phagocytosis activity, oxidative burst activity and cytokine transcription (IL-1β, TNFα, IL-10) were examined, accompanied by in silico modelling. The immunotoxicants dexamethasone, benzo(a)pyrene, ethinylestradiol and bisphenol A significantly altered the immune parameters at non-cytotoxic concentrations whereas diclofenac had only weak effects. However, the two baseline chemicals with no known immunotoxic potential, butanol and ethylene glycol, caused significant effects, too. From our results it appears that the in vitro fish leukocyte assay as performed in the present study has only a limited capacity for discriminating between immunotoxicants and non-immunotoxicants.
Topics: Animals; Benzhydryl Compounds; Benzo(a)pyrene; Butanols; Dexamethasone; Diclofenac; Ethinyl Estradiol; Ethylene Glycol; Female; Fish Proteins; Gene Expression Regulation; Head Kidney; Immunotoxins; Interleukin-10; Interleukin-1beta; Leukocytes; Oncorhynchus mykiss; Phagocytosis; Phenols; Primary Cell Culture; Respiratory Burst; Transcription, Genetic; Tumor Necrosis Factor-alpha; Water Pollutants, Chemical
PubMed: 33542403
DOI: 10.1038/s41598-021-82711-5