zebrafish
Dotaz
Zobrazit nápovědu
After its introduction for scientific investigation in the 1950s, the cypriniform zebrafish, Danio rerio, has become a valuable model for the study of regenerative processes and mechanisms. Zebrafish exhibit epimorphic regeneration, in which a nondifferentiated cell mass formed after amputation is able to fully regenerate lost tissue such as limbs, heart muscle, brain, retina, and spinal cord. The process of limb regeneration in zebrafish comprises several stages characterized by the activation of specific signaling pathways and gene expression. We review current research on key factors in limb regeneration using zebrafish as a model.
- Klíčová slova
- Danio rerio, biomedicine, model organism, regeneration, zebrafish,
- MeSH
- dánio pruhované genetika fyziologie MeSH
- exprese genu * MeSH
- modely u zvířat MeSH
- ploutve zvířat fyziologie MeSH
- regenerace * MeSH
- signální transdukce * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Vertebral bodies are composed of two types of metameric elements, centra and arches, each of which is considered as a developmental module. Most parts of the teleost vertebral column have a one-to-one relationship between centra and arches, although, in all teleosts, this one-to-one relationship is lost in the caudal fin endoskeleton. Deviation from the one-to-one relationship occurs in most vertebrates, related to changes in the number of vertebral centra or to a change in the number of arches. In zebrafish, deviations also occur predominantly in the caudal region of the vertebral column. In-depth phenotypic analysis of wild-type zebrafish was performed using whole-mount stained samples, histological analyses and synchrotron radiation X-ray tomographic microscopy 3D reconstructions. Three deviant centra phenotypes were observed: (i) fusion of two vertebral centra, (ii) wedge-shaped hemivertebrae and (iii) centra with reduced length. Neural and haemal arches and their spines displayed bilateral and unilateral variations that resemble vertebral column phenotypes of stem-ward actinopterygians or other gnathostomes as well as pathological conditions in extant species. Whether it is possible to distinguish variations from pathological alterations and whether alterations resemble ancestral conditions is discussed in the context of centra and arch variations in other vertebrate groups and basal actinopterygian species.
- Klíčová slova
- developmental modules, evolution and development, skeleton, vertebrae, zebrafish,
- MeSH
- dánio pruhované * MeSH
- fenotyp MeSH
- páteř * diagnostické zobrazování MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tuberculosis is still a global health burden. It is caused by Mycobacterium tuberculosis which afflicts around one third of the world's population and costs around 1.3 million people their lives every year. Bacillus Calmette-Guerin vaccine is inefficient to prevent overt infection. Additionally, the lengthy inconvenient course of treatment, along with the raising issue of antimicrobial resistance, result in incomplete eradication of this infectious disease. The lack of proper animal models that replicate the latent and active courses of human tuberculosis infection remains one of the main reasons behind the poor advancement in tuberculosis research. Danio rerio, commonly known as zebrafish, is catching more attention as an animal model in tuberculosis research field. This shift is based on the histological and pathological similarities between Mycobacterium marinum infection in zebrafish and Mycobacterium tuberculosis infection in humans. Being small, cheap, transparent, and easy to handle have added further advantages to the use of zebrafish model. Besides better understanding of the pathogenesis of tuberculosis, Mycobacterium marinum infected zebrafish model is useful for evaluating novel vaccines against human tuberculosis, high throughput small molecule screening, repurposing established drugs with possible antitubercular activity, and assessing novel antituberculars for hepatotoxicity.
- Klíčová slova
- Animal model, drug screening, tuberculosis, vaccine development, zebrafish,
- MeSH
- antituberkulotika farmakologie MeSH
- dánio pruhované * mikrobiologie MeSH
- lidé MeSH
- modely nemocí na zvířatech * MeSH
- Mycobacterium marinum účinky léků fyziologie MeSH
- Mycobacterium tuberculosis účinky léků fyziologie MeSH
- tuberkulóza farmakoterapie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- antituberkulotika MeSH
Macrophages derive from multiple sources of hematopoietic progenitors. Most macrophages require colony-stimulating factor 1 receptor (CSF1R), but some macrophages persist in the absence of CSF1R. Here, we analyzed mpeg1:GFP-expressing macrophages in csf1r-deficient zebrafish and report that embryonic macrophages emerge followed by their developmental arrest. In larvae, mpeg1+ cell numbers then increased showing two distinct types in the skin: branched, putative Langerhans cells, and amoeboid cells. In contrast, although numbers also increased in csf1r-mutants, exclusively amoeboid mpeg1+ cells were present, which we showed by genetic lineage tracing to have a non-hematopoietic origin. They expressed macrophage-associated genes, but also showed decreased phagocytic gene expression and increased epithelial-associated gene expression, characteristic of metaphocytes, recently discovered ectoderm-derived cells. We further demonstrated that juvenile csf1r-deficient zebrafish exhibit systemic macrophage depletion. Thus, csf1r deficiency disrupts embryonic to adult macrophage development. Zebrafish deficient for csf1r are viable and permit analyzing the consequences of macrophage loss throughout life.
Immune cells called macrophages are found in all organs in the body. These cells are highly effective at eating and digesting large particles including dead cells and debris, and microorganisms such as bacteria. Macrophages are also instrumental in shaping developing organs and repairing tissues during life. Macrophages were, until recently, thought to be constantly replenished from cells circulating in the bloodstream. However, it turns out that separate populations of macrophages become established in most tissues during embryonic development and are maintained throughout life without further input. Previous studies of zebrafish, rodents and humans have shown that, when a gene called CSF1R is non-functional, macrophages are absent from many organs including the brain. However, some tissue-specific macrophages still persist, and it was not clear why these cells do not rely on the CSF1R gene while others do. Kuil et al. set out to decipher the precise requirement for the CSF1R gene in macrophage development in living zebrafish. The experiments used zebrafish that make a green fluorescent protein in their macrophages. As these fish are transparent, this meant that Kuil et al. could observe the cells within the living fish and isolate them to determine which genes are switched on and off. This approach revealed that zebrafish with a mutated version of the CSF1R gene make macrophages as embryos but that these cells then fail to multiply and migrate into the developing organs. This results in fewer macrophages in the zebrafish’s tissues, and an absence of these cells in the brain. Kuil et al. went on to show that new macrophages did emerge in zebrafish that were about two to three weeks old. However, unexpectedly, these new cells were not regular macrophages. Instead, they were a new recently identified cell-type called metaphocytes, which share similarities with macrophages but have a completely different origin, move faster and do not eat particles. Zebrafish lacking the CSF1R gene thus lose nearly all their macrophages but retain metaphocytes. These macrophage-free mutant zebrafish constitute an unprecedented tool for further studies looking to discriminate the different roles of macrophages and metaphocytes.
- Klíčová slova
- CSF1R, developmental biology, hematopoiesis, langerhans cells, macrophages, metaphocytes, microglia, zebrafish,
- MeSH
- dánio pruhované embryologie MeSH
- makrofágy metabolismus fyziologie MeSH
- mikroglie metabolismus fyziologie MeSH
- proliferace buněk MeSH
- proteiny dánia pruhovaného metabolismus fyziologie MeSH
- receptory faktoru stimulujícího granulocyto-makrofágové kolonie metabolismus fyziologie MeSH
- stanovení celkové genové exprese MeSH
- tyrosinkinasové receptory MeSH
- tyrosinkinasy metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- audiovizuální média MeSH
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- csf1ra protein, zebrafish MeSH Prohlížeč
- proteiny dánia pruhovaného MeSH
- receptory faktoru stimulujícího granulocyto-makrofágové kolonie MeSH
- tyrosinkinasové receptory MeSH
- tyrosinkinasy MeSH
The aggrandised advancement in utility of advanced day-to-day materials and nanomaterials has raised serious concern on their biocompatibility with human and other biotic members. In last few decades, understanding of toxicity of these materials has been given the centre stage of research using many in vitro and in vivo models. Zebrafish (Danio rerio), a freshwater fish and a member of the minnow family has garnered much attention due to its distinct features, which make it an important and frequently used animal model in various fields of embryology and toxicological studies. Given that fertilization and development of zebrafish eggs take place externally, they serve as an excellent model organism for studying early developmental stages. Moreover, zebrafish possess a comparable genetic composition to humans and share almost 70% of their genes with mammals. This particular model organism has become increasingly popular, especially for developmental research. Moreover, it serves as a link between in vitro studies and in vivo analysis in mammals. It is an appealing choice for vertebrate research, when employing high-throughput methods, due to their small size, swift development, and relatively affordable laboratory setup. This small vertebrate has enhanced comprehension of pathobiology and drug toxicity. This review emphasizes on the recent developments in toxicity screening and assays, and the new insights gained about the toxicity of drugs through these assays. Specifically, the cardio, neural, and, hepatic toxicology studies inferred by applications of nanoparticles have been highlighted.
- Klíčová slova
- Cardiotoxicity, Drug screening, Hepatotoxicity, Nanoparticles, Neurotoxicity, Toxicity, Zebrafish,
- MeSH
- dánio pruhované * MeSH
- játra MeSH
- lidé MeSH
- modely u zvířat MeSH
- nanostruktury * MeSH
- savci MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
We report for the first time, a comparison of two approaches for artificially induced triploidy in zebrafish (Danio rerio) using cold shock and heat shock treatments. Of the two methods, heat shock treatment proved more effective with a triploid production rate of 100% in particular females. Subsequently, triploid zebrafish larvae were used as recipients for intraperitoneal transplantation of ovarian and testicular cells originating from vas:EGFP strain in order to verify their suitability for surrogate reproduction. Production of donor-derived sperm was achieved in 23% of testicular cell recipients and 16% of ovarian cell recipients, indicating the suitability of triploids as surrogate hosts for germ cell transplantation. Success of the transplantation was confirmed by positive GFP signal detected in gonads of dissected fish and stripped sperm. Germline transmission was confirmed by fertilization tests followed by PCR analysis of embryos with GFP specific primers. Reproductive success of germline chimera triploids evaluated as fertilization rate and progeny development was comparable to control groups.
- Klíčová slova
- Chromosome manipulation, Germ cell transplantation, Surrogate reproduction, Triploid, Zebrafish,
- MeSH
- chov metody MeSH
- dánio pruhované genetika MeSH
- genetické inženýrství metody veterinární MeSH
- průtoková cytometrie MeSH
- teplota MeSH
- triploidie * MeSH
- zárodečné buňky transplantace MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
BACKGROUND: Meis family of transcription factors operates in Pbx-Meis-Hox regulatory network controlling development of various tissues including eye, limbs, heart, hindbrain or craniofacial skeletal elements originating from the neural crest. Although studies in mouse provide abundant information about Meis factors function in embryogenesis, little is known about their role in zebrafish. RESULTS: We generated zebrafish lines carrying null mutations in meis1a, meis1b, meis2a, and meis2b genes. Only meis1b mutants are lethal at larval stage around 13 dpf whereas the other mutant lines are viable and fertile. We focused on development of neural crest-derived craniofacial structures such as tendons, cranial nerves, cartilage and accompanying muscles. Meis1b mutants displayed morphogenetic abnormalities in the cartilage originating from the first and second pharyngeal arches. Meckel's cartilage was shorter and wider with fused anterior symphysis and abnormal chondrocyte organization. This resulted in impaired tendons and muscle fiber connections while tenocyte development was not largely affected. CONCLUSIONS: Loss-of-function mutation in meis1b affects cartilage morphology in the lower jaw that leads to disrupted organization of muscles and tendons.
- Klíčová slova
- Meis, chondrocyte differentiation, craniofacial development, muscle fiber, zebrafish,
- MeSH
- chrupavka embryologie metabolismus MeSH
- crista neuralis embryologie metabolismus MeSH
- dánio pruhované * embryologie genetika MeSH
- homeodoménové proteiny * genetika metabolismus MeSH
- lebka * embryologie MeSH
- morfogeneze * genetika fyziologie MeSH
- mutace MeSH
- proteiny dánia pruhovaného * genetika metabolismus MeSH
- transkripční faktor Meis1 MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- homeodoménové proteiny * MeSH
- proteiny dánia pruhovaného * MeSH
- transkripční faktor Meis1 MeSH
Kit ligand (Kitlg) is pleiotropic cytokine with a prominent role in vertebrate erythropoiesis. Although the role of Kitlg in this process has not been reported in Danio rerio (zebrafish), in the present study we show that its function is evolutionarily conserved. Zebrafish possess 2 copies of Kitlg genes (Kitlga and Kitlgb) as a result of whole-genome duplication. To determine the role of each ligand in zebrafish, we performed a series of ex vivo and in vivo gain- and loss-of-function experiments. First, we tested the biological activity of recombinant Kitlg proteins in suspension culture from zebrafish whole-kidney marrow, and we demonstrate that Kitlga is necessary for expansion of erythroid progenitors ex vivo. To further address the role of kitlga and kitlgb in hematopoietic development in vivo, we performed gain-of-function experiments in zebrafish embryos, showing that both ligands cooperate with erythropoietin (Epo) to promote erythroid cell expansion. Finally, using the kita mutant (kitab5/b5 or sparse), we show that the Kita receptor is crucial for Kitlga/b cooperation with Epo in erythroid cells. In summary, using optimized suspension culture conditions with recombinant cytokines (Epo, Kitlga), we report, for the first time, ex vivo suspension cultures of zebrafish hematopoietic progenitor cells that can serve as an indispensable tool to study normal and aberrant hematopoiesis in zebrafish. Furthermore, we conclude that, although partial functional diversification of Kit ligands has been described in other processes, in erythroid development, both paralogs play a similar role, and their function is evolutionarily conserved.
- MeSH
- dánio pruhované MeSH
- erythropoetin * MeSH
- erytroidní buňky MeSH
- ligandy MeSH
- proteiny dánia pruhovaného genetika MeSH
- růstový faktor kmenových buněk genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- erythropoetin * MeSH
- kitlga protein, zebrafish MeSH Prohlížeč
- ligandy MeSH
- proteiny dánia pruhovaného MeSH
- růstový faktor kmenových buněk MeSH
Herbicides are the most widely used group of pesticides but after reaching water bodies they are able to cause adverse effects on non-target organisms. Different formulations using the same active ingredient are frequently available, which raises the issue of potential influence of different formulation types on herbicide toxicity. The present study evaluated the toxicity and teratogenic effects of the active ingredient clomazone and its two formulations (Rampa® EC and GAT Cenit 36 CS, both containing 360g a.i./l of clomazone) on zebrafish embryos. The crucial difference between the two formulation types is the way of active substance release. This investigation is the first report on zebrafish embryotoxicity of both clomazone and its formulations. The technical active ingredient and formulations caused mortality and diverse teratogenic effects, showing different levels of toxicity. The LC50 values for the technical ingredient, Rampa® EC and GAT Cenit 36 CS were 61.4, 9.6 and 92.5mg a.i./l, respectively. Spontaneous movements in 22 hpf embryos decreased under exposure to both the technical ingredient and formulations. A significant number of underdeveloped embryos was detected after exposure to clomazone and Rampa® EC, while no underdevelopment was noted in embryos exposed to GAT Cenit 36 CS. Exposure to the technical ingredient and formulations led also to a series of morphological changes and interfered with the growth of zebrafish embryos. The EC50 based on detection of edemas, spine and tail tip deformations and gas bladder absence (120hpf) was 12.1, 10.1 and 24.1mg/l for technical clomazone, Rampa® EC and GAT Cenit 36 CS, while teratogenicity index (TI) based on LC50/EC50 ratio was 5.1, 1 and 3.8, respectively. The data in this study showed that the emulsifiable concentrate formulation (Rampa® EC) caused statistically significantly higher toxicity, and the aqueous capsule suspension (GAT Cenit 36 CS) lower toxicity than technical clomazone. It indicates that different formulations with the same active ingredient may have different environmental impacts, which is why risk assessment based only on active ingredient toxicity might not be sufficient in terms of preventing formulation effects on the environment.
- Klíčová slova
- Clomazone, Embryo, Embryotoxicity, Formulations, Zebrafish,
- MeSH
- chemické látky znečišťující vodu toxicita MeSH
- dánio pruhované embryologie MeSH
- embryo nesavčí abnormality účinky léků MeSH
- herbicidy toxicita MeSH
- isoxazoly toxicita MeSH
- oxazolidinony toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chemické látky znečišťující vodu MeSH
- clomazone MeSH Prohlížeč
- herbicidy MeSH
- isoxazoly MeSH
- oxazolidinony MeSH
Calcium plays prominent roles in regulating a broad range of physiological events in reproduction. The aim of this study was to describe the subcellular distribution of calcium deposits during stages of oogenesis in zebrafish using a combined oxalate-pyroantimonate technique. The oocyte development of zebrafish was categorized into four stages: primary growth, cortical-alveolus, vitellogenic, and maturation, based on morphological criteria. Calcium deposits in the primary growth stage were detected in the cytoplasm, mitochondria, nucleus, and follicular cells. At the cortical-alveolus stage, calcium particles were transported from follicular cells and deposited in the cortical alveoli. In the vitellogenic stage, some cortical alveoli were compacted and transformed from flocculent electron-lucent to electron-dense objects with the progression of the stage. Calcium deposits were transformed from larger to smaller particles, coinciding with compaction of cortical alveoli. In the maturation stage, calcium deposits in all oocyte compartments decreased, with the exception of those in mitochondria. The proportion of area covered by calcium deposits in the mitochondria and cortical alveoli of oocytes at different stages of development was significantly different (p<0.05). The extent of calcium deposits in the cortical alveoli of mature oocytes was substantially lower than in earlier stages. Basic information about calcium distribution during zebrafish oogenesis may contribute to better understanding of its role in oogenesis.
- Klíčová slova
- Calcium, Follicular cell, Oocyte, Ovary, Oxalate–pyroantimonate, Zebrafish,
- MeSH
- dánio pruhované fyziologie MeSH
- mikroskopie metody MeSH
- oocyty chemie MeSH
- oogeneze * MeSH
- počítačové zpracování obrazu metody MeSH
- vápník analýza MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- vápník MeSH
Understanding the molecular basis of sexual dimorphism in the cardiovascular system may contribute to the improvement of the outcome in biological, pharmacological, and toxicological studies as well as on the development of sex-based drugs and therapeutic approaches. Label-free protein quantification using high-resolution mass spectrometry was applied to detect sex-based proteome differences in the heart of zebrafish Danio rerio. Out of almost 3000 unique identified proteins in the heart, 79 showed significant abundance differences between male and female fish. The functional differences were mapped using enrichment analyses. Our results suggest that a large amount of materials needed for reproduction (e.g., sugars, lipids, proteins, etc.) may impose extra pressure on blood, vessels, and heart on their way toward the ovaries. In the present study, the female's heart shows a clear sexual dimorphism by changing abundance levels of numerous proteins, which could be a way to safely overcome material-induced elevated pressures. These proteins belong to the immune system, oxidative stress response, drug metabolization, detoxification, energy, metabolism, and so on. In conclusion, we showed that sex can induce dimorphism at the molecular level in nonsexual organs such as heart and must be considered as an important factor in cardiovascular research. Data are available via ProteomeXchange with identifier PXD023506.
- Klíčová slova
- cardiovascular system, protein, proteomics, sex-based, zebrafish,
- MeSH
- dánio pruhované * genetika MeSH
- pohlavní dimorfismus * MeSH
- proteiny dánia pruhovaného * MeSH
- proteom genetika MeSH
- proteomika MeSH
- srdce * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- proteiny dánia pruhovaného * MeSH
- proteom MeSH
N-Methyl-d-aspartate receptors (NMDARs) control synaptic plasticity and brain development in a manner determined by receptor subunit composition. Pathogenic variants in GRIN2A gene, encoding the NMDAR GluN2A subunit, can cause gain or loss of function of receptors containing the affected subunit and are associated with intellectual disability and epilepsy in patients. While in vitro studies of recombinant receptors have yielded some insights, animal experimental models are essential to better understand the relationship between the molecular pathology of the variants and the disease. Here we introduce a zebrafish model of GluN2A loss of function to study system-level effects of zebrafish grin2Aa and grin2Ab gene deletion. Our electrophysiological analysis revealed functional differences between receptors containing zebrafish GluN2Aa/b and GluN2Bb paralogs comparable with mammalian receptors containing GluN2A versus GluN2B subunits. Both grin2Aa-/- and grin2Ab-/- as well as double-knock-out grin2A-/- zebrafish larvae showed increased locomotor activity in a novel environment. Proteomic analysis suggested that the relative proportion of GluN2B-containing NMDARs may be increased in grin2A mutant fish. Our results highlight fundamental similarities between zebrafish and mammalian NMDAR signaling and validate the use of zebrafish as a model organism to study the neurodevelopmental role of NMDARs. The newly created transgenic zebrafish strains complement the rodent models of GluN2A loss of function and can be used for high-throughput testing of pharmacological or genetic treatment strategies for patients with GRIN2A gene variants.
- Klíčová slova
- CRISPR-Cas9, Grin genes, channelopathy, glutamate receptors, zebrafish,
- MeSH
- dánio pruhované MeSH
- epilepsie * genetika patofyziologie MeSH
- geneticky modifikovaná zvířata MeSH
- modely nemocí na zvířatech MeSH
- plavání * fyziologie MeSH
- proteiny dánia pruhovaného * genetika MeSH
- receptory N-methyl-D-aspartátu * genetika nedostatek MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- N-methyl D-aspartate receptor subtype 2A MeSH Prohlížeč
- NR2B NMDA receptor MeSH Prohlížeč
- proteiny dánia pruhovaného * MeSH
- receptory N-methyl-D-aspartátu * MeSH
Recently, the indiscriminate use of antibiotics in the aquaculture sector has raised public concern because of possible toxic effects, development of bacterial resistance, and accumulation of residues in individual tissues. Even if several countries have developed regulations about their use, it is clear that long-term growth of the aquaculture industry requires both ecologically sound practices and sustainable resource management. Alternative strategies for better management of antibiotic administration are of primary interest to improve absorption rates and, as a consequence, to reduce their release into the aquatic environment. The present study investigates, for the first time to our knowledge, a new methodology for oxytetracycline (OTC) administration through the use of iron oxide nanoparticles (NPs) (made of maghemite γ-Fe2O3) in zebrafish (Danio rerio). Fish were divided into 4 experimental groups: control; group A exposed to 4 mg/L OTC (through water); group B exposed to the 100 mg/L SAMNs@OTC complex (equivalent to 4 mg/L OTC), and group C exposed to bare NPs. No detoxification processes or anatomical alterations were observed in fish exposed to bare NPs. Exposure of fish to the SAMNs@OTC complex resulted in a 10 times higher OTC accumulation with respect to using water exposure. This new OTC administration method seems much more efficient with respect to the traditional way of exposure and has the potentiality to reduce antibiotic utilization and possible environmental impacts. However, the dynamics related to OTC release from the SAMNs@OTC complex are still not clear and need further investigations.
- Klíčová slova
- antibiotics, aquaculture, detoxification, nanotechnology,
- MeSH
- antibakteriální látky aplikace a dávkování MeSH
- dánio pruhované * MeSH
- kovové nanočástice chemie MeSH
- lékové transportní systémy metody normy MeSH
- oxytetracyklin aplikace a dávkování MeSH
- rybářství MeSH
- železité sloučeniny chemie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibakteriální látky MeSH
- ferric oxide MeSH Prohlížeč
- oxytetracyklin MeSH
- železité sloučeniny MeSH
BACKGROUND: Alpha-Gal syndrome (AGS) is a tick-borne food allergy caused by IgE antibodies against the glycan galactose-alpha-1,3-galactose (α-Gal) present in glycoproteins and glycolipids from mammalian meat. To advance in the diagnosis and treatment of AGS, further research is needed to unravel the molecular and immune mechanisms underlying this syndrome. The objective of this study is the characterization of tick salivary components and proteins with and without α-Gal modifications involved in modulating human immune response against this carbohydrate. METHODS: Protein and α-Gal content were determined in tick saliva components, and proteins were identified by proteomics analysis of tick saliva fractions. Pathophysiological changes were recorded in the zebrafish (Danio rerio) model after exposure to distinct Ixodes ricinus tick salivary components. Serum samples were collected from zebrafish at day 8 of exposure to determine anti-α-Gal, anti-glycan, and anti-tick saliva protein IgM antibody titers by enzyme-linked immunosorbent assay (ELISA). RESULTS: Zebrafish treated with tick saliva and saliva protein fractions combined with non-protein fractions demonstrated significantly higher incidence of hemorrhagic type allergic reactions, abnormal behavioral patterns, or mortality when compared to the phosphate-buffered saline (PBS)-treated control group. The main tick salivary proteins identified in these fractions with possible functional implication in AGS were the secreted protein B7P208-salivary antigen p23 and metalloproteases. Anti-α-Gal and anti-tick salivary gland IgM antibody titers were significantly higher in distinct saliva protein fractions and deglycosylated saliva group when compared with PBS-treated controls. Anti-glycan antibodies showed group-related profiles. CONCLUSIONS: Results support the hypothesis that tick salivary biomolecules with and without α-Gal modifications are involved in modulating immune response against this carbohydrate.
- Klíčová slova
- Allergy, Alpha-gal syndrome, Glycan, Tick, Zebrafish,
- MeSH
- dánio pruhované metabolismus MeSH
- galaktosa MeSH
- imunoglobulin E MeSH
- imunoglobulin M MeSH
- klíště * MeSH
- kousnutí klíštětem * MeSH
- lidé MeSH
- potravinová alergie * etiologie MeSH
- proteiny členovců MeSH
- savci MeSH
- sliny MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- galaktosa MeSH
- imunoglobulin E MeSH
- imunoglobulin M MeSH
- proteiny členovců MeSH
Impaired wound healing is associated with aging and has significant effects on human health on an individual level, but also on the whole health-care sector. Deficient angiogenesis appears to be involved in the process, but the underlying biology is still poorly understood. This is at least partially being explained by complexity and costs in using mammalian aging models. To understand aging-related vascular biology of impaired wound healing, we used zebrafish and turquoise killifish fin regeneration models. The regeneration of caudal fin after resection was significantly reduced in old individuals in both species. Age-related changes in angiogenesis, vascular density and expression levels of angiogenesis biomarker VEGF-A were observed. Furthermore, the anti-angiogenic drug vascular endothelial growth factor receptor blocking inhibitor SU5416 reduced regeneration, indicating a key role for angiogenesis in the regeneration of aging caudal fin despite aging-related changes in vasculature. Taken together, our data indicate that these fish fin regeneration models are suitable for studying aging-related decline in wound healing and associated alterations in aging vasculature.
- Klíčová slova
- Aging, Angiogenesis, Danio rerio, Fin, Nothobranchius furzeri, Regeneration, Turquoise killifish, VEGF, Wound healing, Zebrafish,
- MeSH
- dánio pruhované * metabolismus MeSH
- Fundulidae * MeSH
- hojení ran MeSH
- lidé MeSH
- proteiny dánia pruhovaného MeSH
- savci metabolismus MeSH
- senioři MeSH
- vaskulární endoteliální růstový faktor A metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- proteiny dánia pruhovaného MeSH
- vaskulární endoteliální růstový faktor A MeSH
The zebrafish is a powerful model organism to study the mechanisms governing transition metal ions within whole brain tissue. Zinc is one of the most abundant metal ions in the brain, playing a critical pathophysiological role in neurodegenerative diseases. The homeostasis of free, ionic zinc (Zn2+) is a key intersection point in many of these diseases, including Alzheimer's disease and Parkinson's disease. A Zn2+ imbalance can eventuate several disturbances that may lead to the development of neurodegenerative changes. Therefore, compact, reliable approaches that allow the optical detection of Zn2+ across the whole brain would contribute to our current understanding of the mechanisms that underlie neurological disease pathology. We developed an engineered fluorescence protein-based nanoprobe that can spatially and temporally resolve Zn2+ in living zebrafish brain tissue. The self-assembled engineered fluorescence protein on gold nanoparticles was shown to be confined to defined locations within the brain tissue, enabling site specific studies, compared to fluorescent protein-based molecular tools, which diffuse throughout the brain tissue. Two-photon excitation microscopy confirmed the physical and photometrical stability of these nanoprobes in living zebrafish (Danio rerio) brain tissue, while the addition of Zn2+ quenched the nanoprobe fluorescence. Combining orthogonal sensing methods with our engineered nanoprobes will enable the study of imbalances in homeostatic Zn2+ regulation. The proposed bionanoprobe system offers a versatile platform to couple metal ion specific linkers and contribute to the understanding of neurological diseases.
- Klíčová slova
- fluorescence, gold, nanoparticles, two-photon excitation imaging, zebrafish, zinc,
- MeSH
- dánio pruhované * metabolismus MeSH
- fluorescenční barviva metabolismus MeSH
- ionty metabolismus MeSH
- kovové nanočástice * MeSH
- mozek metabolismus MeSH
- zinek metabolismus MeSH
- zlato metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fluorescenční barviva MeSH
- ionty MeSH
- zinek MeSH
- zlato MeSH
Cyanobacteria routinely release potentially harmful bioactive compounds into the aquatic environment. Several recent studies suggested a potential link between the teratogenicity of effects caused by cyanobacteria and production of retinoids. To investigate this relationship, we analysed the teratogenicity of field-collected cyanobacterial bloom samples by means of an in vivo zebrafish embryo test, an in vitro reporter gene bioassay and by the chemical analysis of retinoids. Extracts of biomass from cyanobacterial blooms with the dominance of Microcystis aeruginosa and Aphanizomenon klebahnii were collected from water bodies in the Czech Republic and showed significant retinoid-like activity in vitro, as well as high degrees of teratogenicity in vivo. Chemical analysis was then used to identify a set of retinoids in ng per gram of dry weight concentration range. Subsequent fractionation and bioassay-based characterization identified two fractions with significant in vitro retinoid-like activity. Moreover, in most of the retinoids eluted from these fractions, teratogenicity with malformations typical for retinoid signalling disruption was observed in zebrafish embryos after exposure to the total extracts and these in vitro effective fractions. The zebrafish embryo test proved to be a sensitive toxicity indicator of the biomass extracts, as the teratogenic effects occurred at even lower concentrations than those expected from the activity detected in vitro. In fact, teratogenicity with retinoid-like activity was detected at concentrations that are commonly found in biomasses and even in bulk water surrounding cyanobacterial blooms. Overall, these results provide evidence of a link between retinoid-like activity, teratogenicity and the retinoids produced by cyanobacterial water blooms in the surrounding environment.
- Klíčová slova
- All-trans retinoic acid, Cyanobacteria, Retinoid-like activity, Retinoids, Teratogenicity, Zebrafish,
- MeSH
- Aphanizomenon patogenita MeSH
- dánio pruhované embryologie genetika MeSH
- embryo nesavčí účinky léků MeSH
- Microcystis patogenita MeSH
- reportérové geny MeSH
- retinoidy biosyntéza toxicita MeSH
- sinice chemie patogenita MeSH
- teratogeny toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- retinoidy MeSH
- teratogeny MeSH
The liver is a central metabolic hub, performing vital functions such as bile production, protein, carbohydrate, lipid and drug metabolism, detoxification of xenobiotics, and the synthesis of essential biomolecules for reproduction, and also shows regenerative capability. Several of these functions can be affected by sexual dimorphisms with important consequences. In this study we used high-throughput proteomics to identify and quantify proteins involved in sexual dimorphism of the zebrafish liver, as a model for preclinical human research. Additionally, we conducted an extensive literature review to explore potential effects of sex-biased protein abundances on liver regeneration capacity and hepatic diseases. The results showed wide-spread sex-specific differences in proteins involved in carbohydrate, protein, and lipid metabolism. Female livers exhibited higher levels of proteins involved in protein synthesis, while male liver protein abundances were higher in energy-producing biochemical pathways, such as the TCA, β-oxidation, and glycolysis. Furthermore, significant sex differences were observed in proteins related to drug metabolism, which should be considered in toxicological and pharmacological research. Some potential links between sex-biased quantities of some key hepatic proteins and the susceptibility of males to liver diseases, as well as the higher hepatic regenerative capacity in females, were suggested. These findings offer a foundation for future targeted research to facilitate the development of sex-specific therapeutic approaches for liver disorders and regenerative medicine. Data are available via ProteomeXchange with identifier PXD061886.
- Klíčová slova
- Drug metabolism, Liver disease, Liver proteomics, Metabolism, Sexual dimorphism, Zebrafish model,
- MeSH
- dánio pruhované * metabolismus MeSH
- játra * metabolismus MeSH
- nemoci jater * metabolismus MeSH
- pohlavní dimorfismus * MeSH
- proteiny dánia pruhovaného * metabolismus MeSH
- proteom metabolismus MeSH
- proteomika metody MeSH
- regenerace jater * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- proteiny dánia pruhovaného * MeSH
- proteom MeSH
This experiment aimed to determine the efficacy of fulvic acid (FLA) on growth performance, innate immune system, antioxidant parameters, and expression of immune and antioxidant-related genes in zebrafish (Danio rerio). To this end, 12 tanks (3 per group), each containing 50 zebrafish (with an average weight of 85.7 ± 10.05 mg) in 72 L, were assigned to diets containing FLA at four levels: 0 (control), 0.25 (FLA1), 0.5 (FLA2), and 1 (FLA3) g/kg diet. Following an eight-week culture period, no significant differences in growth performance were observed among the treatment groups (P > 0.05). However, lysozyme activity, total immunoglobulin (Ig), and total protein concentrations in whole-body extracts were significantly enhanced in the 0.5-1 g FLA/kg diet groups compared to the other treatments (P < 0.05). No significant differences were observed among the groups in catalase (CAT), glutathione peroxidase (GPx), or superoxide dismutase (SOD) activities (P > 0.05). The supplementation of FLA significantly upregulated the gene expression of interferon-α (IFN-α) and tumor necrosis factor-alpha (TNF-α), with the highest expression observed in the 0.5 g FLA/kg diet group (P < 0.05). Additionally, interleukin 1 (IL-1) expression was markedly elevated in this group in comparison to the other treatments (P < 0.05). While there was a significant increase in GPx gene expression with dietary FLA (P < 0.05), no notable differences were observed among FLA treatments (P > 0.05). CAT gene expression remained consistent across all groups (P > 0.05). In contrast, SOD gene expression significantly increased in response to all FLA-supplemented diets, with the highest level observed in the 0.5 g FLA/kg group (P < 0.05). These findings suggest that FLA may serve as an effective dietary supplement to enhance the immune response and antioxidant capacity in zebrafish.
- Klíčová slova
- Antioxidant, Cytokine, Fulvic acid, Growth performance, Innate immunity, Zebrafish,
- MeSH
- antioxidancia * metabolismus MeSH
- benzopyrany * farmakologie MeSH
- dánio pruhované * imunologie růst a vývoj metabolismus MeSH
- krmivo pro zvířata MeSH
- potravní doplňky MeSH
- přirozená imunita * účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antioxidancia * MeSH
- benzopyrany * MeSH
- fulvic acid MeSH Prohlížeč
Neurofibromatosis type 2 (NF-2) is a dominantly inherited genetic disorder that results from variants in the tumor suppressor gene, neurofibromin 2 (NF2). Here, we report the generation of a conditional zebrafish model of neurofibromatosis established by inducible genetic knockout of nf2a/b, the zebrafish homologs of human NF2. Analysis of nf2a and nf2b expression revealed ubiquitous expression of nf2b in the early embryo, with overlapping expression in the neural crest and its derivatives and in the cranial mesenchyme. In contrast, nf2a displayed lower expression levels. Induction of nf2a/b knockout at early stages increased the proliferation of larval Schwann cells and meningeal fibroblasts. Subsequently, in adult zebrafish, nf2a/b knockout triggered the development of a spectrum of tumors, including vestibular Schwannomas, spinal Schwannomas, meningiomas and retinal hamartomas, mirroring the tumor manifestations observed in patients with NF-2. Collectively, these findings highlight the generation of a novel zebrafish model that mimics the complexities of the human NF-2 disorder. Consequently, this model holds significant potential for facilitating therapeutic screening and elucidating key driver genes implicated in NF-2 onset.
- Klíčová slova
- Cancer, Inducible knockout, Meninges, Neurofibromatosis type 2, Schwann cells, Zebrafish,
- MeSH
- dánio pruhované * genetika embryologie MeSH
- geneticky modifikovaná zvířata MeSH
- genový knockout * MeSH
- larva metabolismus MeSH
- lidé MeSH
- modely nemocí na zvířatech * MeSH
- neurofibromatóza 2 genetika patologie metabolismus MeSH
- neurofibromatózy genetika patologie metabolismus MeSH
- neurofibromin 2 * genetika metabolismus nedostatek MeSH
- proliferace buněk MeSH
- proteiny dánia pruhovaného * genetika metabolismus nedostatek MeSH
- Schwannovy buňky metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- neurofibromin 2 * MeSH
- proteiny dánia pruhovaného * MeSH
Ziram is a broad spectrum pesticide that belongs to the class of dimethyl-dithiocarbamate (DTC) fungicides. The objectives of this study were to assess the effects of ziram in developing zebrafish. Ziram was highly toxic to zebrafish embryos, with a 96-h LC50 value of 1082.54 nM (∼0.33 mg/L). Zebrafish embryos at 6 h post-fertilization (hpf) were exposed to solvent control (0.1% DMSO), or one dose of 1, 10, 100, and 1000 nM ziram for 96 h. Ziram induced lethality in a dose-dependent manner, decreased hatching rate and heartbeat, and caused wavy deformities at 72 and 96 hpf at 100 and 1000 nM. Basal oxygen consumption rates of zebrafish at 24 hpf were decreased with 1000 nM, suggesting that ziram affects oxidative phosphorylation. We also measured the expression of transcripts associated with the oxidative stress response (sod1 and sod2) and dopamine receptor signaling at ∼96 h of exposure. There was no difference in the expression of genes related to oxidative stress, nor those related to the dopamine system. Locomotor activity was also assessed in larval zebrafish (7 dpf), and ziram increased total activity, the velocity in light zone, and total distance moved at 10 nM, while it decreased the mean time spent in the dark zone at 1 and 10 nM. Behavioral responses were dependent upon the time point and clutch examined. These data demonstrate that ziram negatively impacts embryonic development (i.e. mortality, hatching, heartbeat and notochord development) of zebrafish, decreases basal respiration of embryos, and alters behavioral responses in larvae.
- Klíčová slova
- Dark/light preference, Dopamine system, Gene expression, Mitochondrial bioenergetics, Zebrafish embryos, Ziram,
- MeSH
- chování zvířat účinky léků MeSH
- dánio pruhované růst a vývoj metabolismus MeSH
- dopamin genetika MeSH
- embryo nesavčí účinky léků MeSH
- embryonální vývoj účinky léků MeSH
- larva účinky léků MeSH
- lokomoce účinky léků MeSH
- oxidační stres genetika MeSH
- průmyslové fungicidy metabolismus toxicita MeSH
- spotřeba kyslíku účinky léků MeSH
- ziram toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dopamin MeSH
- průmyslové fungicidy MeSH
- ziram MeSH
Many reports on anti-progestogenic activities in aquatic environments have been published in the past decade. These are monitored mainly by in vitro reporter gene bioassays based upon the human progesterone receptor (PR). However, results obtained by some human in vitro bioassays may not be relevant for aquatic animals, especially fish. The present work aimed to detect fish (anti-)PR activity in waste- and receiving surface waters. In parallel, human (anti-)PR activity was analysed to determine if there was any connection between human and fish (anti-)PR activities. Finally, (anti-)PR activities were linked to the occurrence of progestins in water samples. Human PR agonistic activity was detected in all wastewater and most receiving surface water samples. Nevertheless, zebrafish PR (zfPR) agonistic activity was found in only two influent wastewater samples (max. 117 ng/L 17α,20β-dihydroxy-4-pregnen-3-one [DHP] equivalents). Analysed synthetic progestins and progesterone accounted for 14 % to 161 % of detected human PR (hPR) agonistic activity in water samples. Progesterone also contributed significantly to zfPR agonistic activity (up to 10 %) in raw wastewater. The anti-hPR activity was detected also in most wastewater and some surface water samples, but synthetic progestins did not trigger anti-zfPR activity in excess of LOQ values. In addition, altrenogest, dienogest, and ulipristal acetate were tested for their potency to zfPR for the first time. The activity analyses of both pure substances and environmental samples showed that human and zebrafish progesterone receptors are differentially activated. Therefore, results based on human PR in vitro bioassays could not predict fish PR activities in the environment.
- Klíčová slova
- Human, In vitro bioassay, Progesterone receptor, Progestin, Wastewater, Zebrafish,
- MeSH
- dánio pruhované * MeSH
- lidé MeSH
- odpadní voda MeSH
- progesteron MeSH
- progestiny analýza MeSH
- receptory progesteronu * MeSH
- voda analýza MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- odpadní voda MeSH
- progesteron MeSH
- progestiny MeSH
- receptory progesteronu * MeSH
- voda MeSH
Notch signalling is critical for the development of the nervous system. In the zebrafish mindbomb mutants, disruption of E3 ubiquitin ligase activity inhibits Notch signalling. In these mutant embryos, precocious development of primary neurons leading to depletion of neural progenitor cells results in a neurogenic phenotype characterized by defects in neural patterning and brain development. Cyclin-dependent kinase 5 (Cdk5), a predominant neuronal kinase, is involved in a variety of essential functions of the nervous system. Most recently, mammalian studies on Notch and Cdk5 regulating each other's function have been emerging. The status of Cdk5 in the mindbomb mutant embryos with excessive primary neurons is not known. In situ hybridization of the zebrafish mindbomb mutant embryos uncovered a robust upregulation in Cdk5 expression but with a reduced Cdk5 activity. The implications of these findings in both the mammalian system and zebrafish are discussed in this mini-review to provide a glimpse into the relationship between Notch and Cdk5 that may explain certain neurodevelopmental defects associated with either mutations in ubiquitin ligase or altered expression of Cdk5.
- MeSH
- biologické modely MeSH
- cyklin-dependentní kinasa 5 metabolismus MeSH
- dánio pruhované metabolismus MeSH
- mutace genetika MeSH
- proteiny dánia pruhovaného genetika MeSH
- receptory Notch metabolismus MeSH
- upregulace genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- cyklin-dependentní kinasa 5 MeSH
- proteiny dánia pruhovaného MeSH
- receptory Notch MeSH
DAF-FM DA is widely used as a live staining compound to show the presence of nitric oxide (NO) in cells. Applying this stain to live zebrafish embryos is known to indicate early centers of bone formation, but the precise (cellular) location of the signal has hitherto not been revealed. Using sections of zebrafish embryos live-stained with DAF-FM DA, we could confirm that the fluorescent signals were predominantly located in areas of ongoing bone formation. Signals were observed in the bone and tooth matrix, in the notochord sheath, as well as in the bulbus arteriosus. Surprisingly, however, they were exclusively extracellular, even after very short staining times. Von Kossa and Alizarin red S staining to reveal mineral deposits showed that DAF-FM DA stains both the mineralized and non-mineralized bone matrix (osteoid), excluding that DAF-FM DA binds non-specifically to calcified structures. The importance of NO in bone formation by osteoblasts is nevertheless undisputed, as shown by the absence of bone structures after the inhibition of NOS enzymes that catalyze the formation of NO. In conclusion, in zebrafish skeletal biology, DAF-FM DA is appropriate to reveal bone formation in vivo, independent of mineralization of the bone matrix, but it does not demonstrate intracellular NO.
- Klíčová slova
- bulbus arteriosus, nitric oxide, notochord sheath, ossification, osteoblasts, zebrafish,
- MeSH
- barvení a značení MeSH
- barvicí látky metabolismus MeSH
- dánio pruhované * metabolismus MeSH
- kosti a kostní tkáň metabolismus MeSH
- osteogeneze * MeSH
- oxid dusnatý metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- barvicí látky MeSH
- oxid dusnatý MeSH
OBJECTIVES: Xenoestrogenic potential of propylparaben (PP), one of the most commonly used preservatives in drugs, cosmetics and food, was investigated in vivo using zebrafish (Danio rerio). METHODS: Juvenile zebrafish (20 days post hatching) were exposed to three different concentrations of propylparaben (PP) dissolved in ethanol and added into the water. After 20 days of exposure the fish were euthanized and vitellogenin concentrations in their whole body homogenates were measured by the method of direct sandwich ELISA. Simultaneously, vitellogenin concentrations in either fish from the control group (exposed to solvent without the substance tested) and in fish from the positive control group (exposed to natural estrogen 17beta-estradiol) were measured. RESULTS: Vitellogenin concentration in whole body homogenates of control fish was 400 (396-540) ng/ml(-1) (geometric mean (95% CI)). Zebrafish exposure to propylparaben at the concentrations of 0.1; 0.4 and 0.9 mg/l(-1) elicited statistically significant decline (P<0.001) of vitellogenin production, i.e. geometric means of vitellogenin concentrations in whole body homogenates were 240 (186-311); 218 (175-270) and 270 (234-311) ng/ml(-1), respectively. Conversely, the geometric mean of vitellogenin concentration in whole body homogenates of zebrafish exposed to 100 ng/ml(-1) of 17beta-estradiol (positive control) was significantly higher (P<0.001) than values in all other groups, i.e. 35,553 (16,860-74,968) ng/ml(-1). CONCLUSIONS: Our results suggest an antiestrogenic potential of propylparaben tested in vivo in juvenile zebrafish (Danio rerio). The estrogenic effect of 17beta-estradiol was confirmed.
- MeSH
- dánio pruhované * MeSH
- estrogeny toxicita MeSH
- konzervační prostředky farmaceutické toxicita MeSH
- parabeny toxicita MeSH
- testy toxicity MeSH
- vitelogeniny analýza MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- srovnávací studie MeSH
- Názvy látek
- estrogeny MeSH
- konzervační prostředky farmaceutické MeSH
- parabeny MeSH
- propylparaben MeSH Prohlížeč
- vitelogeniny MeSH
This protocol describes the ex vivo characterization of zebrafish hematopoietic progenitors. We show how to isolate zebrafish hematopoietic cells for cultivation and differentiation in colony assays in semi-solid media. We also describe procedures for the generation of recombinant zebrafish cytokines and for the isolation of carp serum, which are essential components of the medium required to grow zebrafish hematopoietic cells ex vivo. The outcome of these clonal assays can easily be evaluated using standard microscopy techniques after 3-10 d in culture. In addition, we describe how to isolate individual colonies for further imaging and gene expression profiling. In other vertebrate model organisms, ex vivo assays have been crucial for elucidating the relationships among hematopoietic stem cells (HSCs), progenitor cells and their mature progeny. The present protocol should facilitate such studies on cells derived from zebrafish.
- MeSH
- buněčné kultury MeSH
- cytokiny genetika MeSH
- dánio pruhované krev MeSH
- hematopoetické kmenové buňky cytologie MeSH
- hematopoéza * MeSH
- kapři krev MeSH
- kultivační média chemie MeSH
- molekulární biologie metody MeSH
- proteiny dánia pruhovaného genetika MeSH
- rekombinantní proteiny genetika MeSH
- stanovení celkové genové exprese MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- cytokiny MeSH
- kultivační média MeSH
- proteiny dánia pruhovaného MeSH
- rekombinantní proteiny MeSH
Alzheimer's disease (AD) is a progressive, fatal, neurodegenerative disorder for which only treatments of limited efficacy are available. Despite early mentions of dementia in the ancient literature and the first patient diagnosed in 1906, the underlying causes of AD are not well understood. This study examined the possible role of dopamine, a neurotransmitter that is involved in cognitive and motor function, in AD. We treated adult zebrafish (Danio rerio) with okadaic acid (OKA) to model AD and assessed the resulting behavioral and neurochemical changes. We then employed a latent learning paradigm to assess cognitive and motor function followed by neurochemical analysis with fast-scan cyclic voltammetry (FSCV) at carbon fiber microelectrodes to measure the electrically stimulated dopamine release. The behavioral assay showed that OKA treatment caused fish to have lower motivation to reach the goal chamber, resulting in impeded learning and decreased locomotor activity compared to controls. Our voltammetric measurements revealed that the peak dopamine overflow in OKA-treated fish was about one-third of that measured in controls. These findings highlight the profound neurochemical changes that may occur in AD. Furthermore, they demonstrate that applying the latent learning paradigm and FSCV to zebrafish is a promising tool for future neurochemical studies and may be useful for screening drugs for the treatment of AD.
- Klíčová slova
- Alzheimer’s disease, behavior, dopamine, fast-scan cyclic voltammetry, okadaic acid, zebrafish,
- MeSH
- Alzheimerova nemoc * MeSH
- dánio pruhované MeSH
- dopamin * MeSH
- karbonové vlákno MeSH
- kyselina okadaová MeSH
- mikroelektrody MeSH
- neurotransmiterové látky MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- dopamin * MeSH
- karbonové vlákno MeSH
- kyselina okadaová MeSH
- neurotransmiterové látky MeSH
In the last decade, zebrafish have accompanied the mouse as a robust animal model for cancer research. The possibility of screening small-molecule inhibitors in a large number of zebrafish embryos makes this model particularly valuable. However, the dynamic visualization of fluorescently labeled tumor cells needs to be complemented by a more sensitive, easy, and rapid mode for evaluating tumor growth in vivo to enable high-throughput screening of clinically relevant drugs. In this study we proposed and validated a pre-clinical screening model for drug discovery by utilizing bioluminescence as our readout for the determination of transplanted cancer cell growth and inhibition in zebrafish embryos. For this purpose, we used NanoLuc luciferase, which ensured rapid cancer cell growth quantification in vivo with high sensitivity and low background when compared to conventional fluorescence measurements. This allowed us large-scale evaluation of in vivo drug responses of 180 kinase inhibitors in zebrafish. Our bioluminescent screening platform could facilitate identification of new small-molecules for targeted cancer therapy as well as for drug repurposing.
- Klíčová slova
- bioluminescence, cancer, high-throughput screening, inhibitor, xenotransplantation, zebrafish,
- Publikační typ
- časopisecké články MeSH
Calcium plays a variety of vital regulatory functions in many physiological and biochemical events in the cell. The aim of this study was to describe the ultrastructural distribution of calcium during different developmental stages of spermatogenesis in a model organism, the zebrafish (Danio rerio), using a combined oxalate-pyroantimonate technique. Samples were treated by potassium oxalate and potassium pyroantimonate during two fixation stages and examined using transmission electron microscopy to detect electron dense intracellular calcium. The subcellular distribution of intracellular calcium was characterized in spermatogonium, spermatocyte, spermatid, and spermatozoon stages. The area which is covered by intracellular calcium in different stages was quantified and compared using software. Isolated calcium deposits were mainly detectable in the cytoplasm and the nucleus of the spermatogonium and spermatocyte. In the spermatid, calcium was partially localized in the cytoplasm as isolated deposits. However, most calcium was transformed from isolated deposits into an unbound pool (free calcium) within the nucleus of the spermatid and the spermatozoon. Interestingly, in the spermatozoon, calcium was mainly localized in a form of an unbound pool which was detectable as an electron-dense mass within the nucleus. Also, sporadic calcium deposits were scattered in the midpiece and flagellum. The proportional area which was covered by intracellular calcium increased significantly from early to late stages of spermatogenesis. The extent of the area which was covered by intracellular calcium in the spermatozoon was the highest compared to earlier stages. Calcium deposits were also observed in the somatic cells (Sertoli, myoid, Leydig) of zebrafish testis. The notable changes in the distribution of intracellular calcium of germ cells during different developmental stages of zebrafish spermatogenesis suggest its different homeostasis and physiological functions during the process of male gamete development.
- Klíčová slova
- electron microscopy, oxalate-pyroantimonate, quantification, testis, ultrastructural localization,
- MeSH
- buněčné jádro ultrastruktura MeSH
- dánio pruhované metabolismus MeSH
- spermatidy cytologie ultrastruktura MeSH
- spermatogeneze * MeSH
- subcelulární frakce metabolismus ultrastruktura MeSH
- testis cytologie ultrastruktura MeSH
- vápník metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- vápník MeSH
In nonmammalian vertebrates, the functional units of hemostasis are thrombocytes. Thrombocytes are thought to arise from bipotent thrombocytic/erythroid progenitors (TEPs). TEPs have been experimentally demonstrated in avian models of hematopoiesis, and mammals possess functional equivalents known as megakaryocyte/erythroid progenitors (MEPs). However, the presence of TEPs in teleosts has only been speculated. To identify and prospectively isolate TEPs, we identified, cloned, and generated recombinant zebrafish thrombopoietin (Tpo). Tpo mRNA expanded itga2b:GFP(+) (cd41:GFP(+)) thrombocytes as well as hematopoietic stem and progenitor cells (HSPCs) in the zebrafish embryo. Utilizing Tpo in clonal methylcellulose assays, we describe for the first time the prospective isolation and characterization of TEPs from transgenic zebrafish. Combinatorial use of zebrafish Tpo, erythropoietin, and granulocyte colony stimulating factor (Gcsf) allowed the investigation of HSPCs responsible for erythro-, myelo-, and thrombo-poietic differentiation. Utilizing these assays allowed the visualization and differentiation of hematopoietic progenitors ex vivo in real-time with time-lapse and high-throughput microscopy, allowing analyses of their clonogenic and proliferative capacity. These studies indicate that the functional role of Tpo in the differentiation of thrombocytes from HSPCs is well conserved among vertebrate organisms, positing the zebrafish as an excellent model to investigate diseases caused by dysregulated erythro- and thrombo-poietic differentiation.
- MeSH
- buněčná diferenciace MeSH
- dánio pruhované embryologie fyziologie MeSH
- embryo nesavčí MeSH
- geneticky modifikovaná zvířata MeSH
- hematopoetické kmenové buňky fyziologie MeSH
- hematopoéza genetika MeSH
- kultivované buňky MeSH
- proliferace buněk MeSH
- thrombopoetin genetika MeSH
- trombocyty fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- thrombopoetin MeSH
Oxytocin is a nonapeptide hormone involved in numerous physiological functions. Real-time electrochemical measurements of oxytocin in living tissue are challenging due to electrode fouling and the large potentials needed to oxidize the tyrosine residue. Here, we used fast-scan cyclic voltammetry at carbon-fiber microelectrodes and flow injection analysis to optimize a waveform for the measurement of oxytocin. This optimized waveform employed an accumulation potential of -0.6 V, multiple scan rates, and a 3 ms holding potential at a positive, oxidizing potential of +1.4 V before linearly scanning the potential back to -0.6 V (versus Ag/AgCl). We obtained a limit of quantitation of 0.34 ± 0.02 μM, and our electrodes did not foul upon multiple injections. Moreover, to demonstrate the utility of our method, we measured the release of oxytocin, evoked by light application and mechanical perturbation, in whole brains from genetically engineered adult zebrafish that express channelrhodopsin-2 selectively on oxytocinergic neurons. Collectively, this work expands the toolkit for the measurement of peptides in living tissue preparations.
- MeSH
- dánio pruhované * MeSH
- karbonové vlákno MeSH
- mikroelektrody MeSH
- neurony MeSH
- oxytocin * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- karbonové vlákno MeSH
- oxytocin * MeSH
Retinoids are newly detected compounds in aquatic ecosystems associated with cyanobacterial water blooms. Their potential health risks are only scarcely described despite numerous detections of all-trans retinoic acid (ATRA) and its derivatives in the environment. Besides the known teratogen ATRA there is only little or no information about their potency and namely their effects in vivo. We characterize ATRA and 8 other retinoids reported to occur in the environment for their bioactivity and teratogenicity using four in vitro reporter gene assays and zebrafish (Danio rerio) embryotoxicity assay. Our results document the ability of these compounds to interfere with retinoid signalling and cause teratogenicity at environmentally relevant levels with EC50 values at nM (hundreds of ng/L) levels and teratogenic indexes ranging from 2.8 (9cis retinoic acid) to 15.8 (retinal). The relative potency of individual compounds for teratogenicity ranged from 0.059 (retinal) to 0.96 (5,6-epoxy ATRA) when compared to ATRA. An environmentally relevant mixture of retinoids was tested showing good predictability of teratogenicity from the in vitro activities and additive toxicity of the mixture. The high teratogenicity of the newly described compounds associated with cyanobacteria presents a concern for developmental stages due to high conservation of the retinoid signalling across vertebrates.
- Klíčová slova
- In vitro, RAR, Retinoids, Teratogenicity, Zebrafish,
- MeSH
- chemické látky znečišťující vodu * toxicita MeSH
- dánio pruhované genetika MeSH
- ekosystém MeSH
- Microcystis * MeSH
- retinoidy toxicita MeSH
- sinice * MeSH
- teratogeny toxicita MeSH
- tretinoin toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chemické látky znečišťující vodu * MeSH
- retinoidy MeSH
- teratogeny MeSH
- tretinoin MeSH
Carbendazim is a widely used broad spectrum benzimidazole fungicide; however, its effects to non-target aquatic organisms are poorly studied. The aim of this study was to investigate the toxic effects of carbendazim to zebrafish early life stages at several levels of biological organization, including developmental, biochemical and behavioural levels. The embryo assay was done following the OECD guideline 236 and using a concentration range between 1.1 and 1.8mg/L. Lethal and developmental endpoints such as hatching, edemas, malformations, heart beat rate, body growth and delays were assessed in a 96h exposure. A sub-teratogenic range (from 0.16 to 500μg/L) was then used to assess effects at biochemical and behavioural levels. Biochemical markers included cholinesterase (ChE), glutathione-S-transferase (GST), lactate dehydrogenase (LDH) and catalase (CAT) and were assessed at 96h. The locomotor behaviour was assessed using an automated video tracking system at 120h. Carbendazim (96h-LC50 of 1.75mg/L) elicited several developmental anomalies in zebrafish embryos with EC50 values ranging from 0.85 to 1.6mg/L. ChE, GST and LDH activities were increased at concentrations equal or above 4μg/L. The locomotor assay showed to be extremely sensitive, detecting effects in time that larvae spent swimming at concentrations of 0.16μg/L and thus, being several orders of magnitude more sensitive that developmental parameters or lethality. These are ecological relevant concentrations and highlight the potential of behavioural endpoints as early warning signs for environmental stress. Further studies should focus on understanding how the behavioural disturbances measured in these types of studies translate into fitness impairment at the adult stage.
- Klíčová slova
- Biomarkers, Locomotor response, Sublethal, Swimming behaviour, Zebrafish,
- MeSH
- benzimidazoly analýza toxicita MeSH
- chemické látky znečišťující vodu analýza toxicita MeSH
- cholinesterasy metabolismus MeSH
- chování zvířat účinky léků MeSH
- dánio pruhované růst a vývoj fyziologie MeSH
- embryo nesavčí účinky léků fyziologie MeSH
- glutathiontransferasa metabolismus MeSH
- karbamáty analýza toxicita MeSH
- katalasa metabolismus MeSH
- larva účinky léků fyziologie MeSH
- lokomoce účinky léků MeSH
- plavání MeSH
- průmyslové fungicidy analýza toxicita MeSH
- tandemová hmotnostní spektrometrie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- benzimidazoly MeSH
- carbendazim MeSH Prohlížeč
- chemické látky znečišťující vodu MeSH
- cholinesterasy MeSH
- glutathiontransferasa MeSH
- karbamáty MeSH
- katalasa MeSH
- průmyslové fungicidy MeSH
Chemical exposure during the early life stages of development may have long lasting effects on organisms that are rarely studied. The present work intended to evaluate the effect of embryonic exposure to the pesticide carbaryl on adult fish behavior. Zebrafish (Danio rerio) embryos were exposed, for 4 days, to sublethal concentrations of carbaryl (0.01, 0.1 and 1.0 mg/L) plus a control and then kept in standard cultivation conditions until adulthood. A battery of behavioral tests was then performed to assess anxiety-like behavior (locomotor activity, thigmotaxis and novel tank diving test), social behavior, and feeding. Developmental exposure of zebrafish to sublethal concentrations of carbaryl produced important behavioral alterations in the adulthood. Main effects included decreased locomotion/hypoactivity (increase in slow movements and decrease of medium and rapid movements), especially in the light periods. Moreover, spatial pattern also changed: while during dark periods control fish increased activity in the outer zone of the tank, this was not observed in exposed fish. Overall, this demonstrated the importance of life stage exposure, clearly demonstrating long lasting effects of a (chemical) stress event at embryonic stages. This data supports the need of considering this scenario in environmental risk evaluations. Further work should focus on the mechanistic effects of developmental disruption responsible for the effects observed.
- Klíčová slova
- Embryonic exposure, Feeding, Locomotion, Long-term effects, Pesticides,
- MeSH
- chování zvířat účinky léků MeSH
- dánio pruhované embryologie MeSH
- embryo nesavčí účinky léků MeSH
- insekticidy toxicita MeSH
- karbaryl toxicita MeSH
- lokomoce účinky léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- insekticidy MeSH
- karbaryl MeSH
The effect of venlafaxine, a pharmaceutical commonly found in aquatic environment, was analyzed on non-target organism, Danio rerio (Hamilton, 1822). D. rerio embryos were treated by two different concentrations of venlafaxine: either concentration relevant in aquatic environment (0.3 μg/L) or concentration that was two orders of magnitude higher (30 μg/L) for the evaluation of dose-dependent effect. Time-dependent effect was rated at 24, 96, and 144 h post-fertilization (hpf). For gene expression, genes representing one of the phases of xenobiotic biotransformation (0 to III) were selected. The results of this study showed that the effect of venlafaxine on the zebrafish embryos is the most evident at hatching (96 hpf). At this time, the results showed a downregulation of gene expression in each phase of biotransformation and in both tested concentrations. In contrast, an upregulation of most of the genes was observed 144 hpf for both tested venlafaxine concentrations. The study shows that venlafaxine can affect the gene expression of biotransformation enzymes in D. rerio embryos even in the environmentally relevant concentration and thus disrupt the process of biotransformation. Moreover, the pxr regulation of genes seems to be disrupted after venlafaxine exposure in dose- and time-dependent manner.
- Klíčová slova
- : ABC transporters, Metabolism, Pharmaceutical, Regulation, Xenobiotics, Zebrafish, pxr,
- MeSH
- antidepresiva farmakologie MeSH
- biotransformace MeSH
- chemické látky znečišťující vodu farmakologie MeSH
- dánio pruhované * MeSH
- embryo nesavčí účinky léků enzymologie MeSH
- regulace genové exprese enzymů * MeSH
- venlafaxin hydrochlorid farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antidepresiva MeSH
- chemické látky znečišťující vodu MeSH
- venlafaxin hydrochlorid MeSH
Zebrafish (Danio rerio) is a valuable non-mammalian vertebrate model widely used to study development and disease, including more recently cancer. The evolutionary conservation of cancer-related programs between human and zebrafish is striking and allows extrapolation of research outcomes obtained in fish back to humans. Zebrafish has gained attention as a robust model for cancer research mainly because of its high fecundity, cost-effective maintenance, dynamic visualization of tumor growth in vivo, and the possibility of chemical screening in large numbers of animals at reasonable costs. Novel approaches in modeling tumor growth, such as using transgene electroporation in adult zebrafish, could improve our knowledge about the spatial and temporal control of cancer formation and progression in vivo. Looking at genetic as well as epigenetic alterations could be important to explain the pathogenesis of a disease as complex as cancer. In this review, we highlight classic genetic and transplantation models of cancer in zebrafish as well as provide new insights on advances in cancer modeling. Recent progress in zebrafish xenotransplantation studies and drug screening has shown that zebrafish is a reliable model to study human cancer and could be suitable for evaluating patient-derived xenograft cell invasiveness. Rapid, large-scale evaluation of in vivo drug responses and kinetics in zebrafish could undoubtedly lead to new applications in personalized medicine and combination therapy. For all of the above-mentioned reasons, zebrafish is approaching a future of being a pre-clinical cancer model, alongside the mouse. However, the mouse will continue to be valuable in the last steps of pre-clinical drug screening, mostly because of the highly conserved mammalian genome and biological processes.
- Klíčová slova
- zebrafish, epigenetics, xenotransplantation, drug screen, pre-clinical cancer model,
- MeSH
- dánio pruhované genetika MeSH
- druhová specificita MeSH
- epigeneze genetická MeSH
- geneticky modifikovaná zvířata genetika MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- mutace MeSH
- myši genetika MeSH
- nádorové biomarkery genetika MeSH
- nádorové buněčné linie MeSH
- nádory genetika MeSH
- regulace genové exprese u nádorů * MeSH
- sekvenování celého genomu MeSH
- technika přenosu genů MeSH
- xenogenní modely - testy protinádorové aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši genetika MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- nádorové biomarkery MeSH
Emerging environmental contaminants, organophosphate flame retardants (OPFRs), pose significant threats to ecosystems and human health. Despite numerous studies reporting the toxic effects of OPFRs, research on their epigenetic alterations remains limited. In this study, we investigated the effects of exposure to 2-ethylhexyl diphenyl phosphate (EHDPP), tricresyl phosphate (TMPP), and triphenyl phosphate (TPHP) on DNA methylation patterns during zebrafish embryonic development. We assessed general toxicity and morphological changes, measured global DNA methylation and hydroxymethylation levels, and evaluated DNA methyltransferase (DNMT) enzyme activity, as well as mRNA expression of DNMTs and ten-eleven translocation (TET) methylcytosine dioxygenase genes. Additionally, we analyzed genome-wide methylation patterns in zebrafish larvae using reduced-representation bisulfite sequencing. Our morphological assessment revealed no general toxicity, but a statistically significant yet subtle decrease in body length following exposure to TMPP and EHDPP, along with a reduction in head height after TPHP exposure, was observed. Eye diameter and head width were unaffected by any of the OPFRs. There were no significant changes in global DNA methylation levels in any exposure group, and TMPP showed no clear effect on DNMT expression. However, EHDPP significantly decreased only DNMT1 expression, while TPHP exposure reduced the expression of several DNMT orthologues and TETs in zebrafish larvae, leading to genome-wide aberrant DNA methylation. Differential methylation occurred primarily in introns (43%) and intergenic regions (37%), with 9% and 10% occurring in exons and promoter regions, respectively. Pathway enrichment analysis of differentially methylated region-associated genes indicated that TPHP exposure enhanced several biological and molecular functions corresponding to metabolism and neurological development. KEGG enrichment analysis further revealed TPHP-mediated potential effects on several signaling pathways including TGFβ, cytokine, and insulin signaling. This study identifies specific changes in DNA methylation in zebrafish larvae after TPHP exposure and brings novel insights into the epigenetic mode of action of TPHP.
- MeSH
- dánio pruhované * genetika růst a vývoj metabolismus MeSH
- larva * účinky léků genetika metabolismus růst a vývoj MeSH
- metylace DNA * účinky léků MeSH
- organofosfáty * toxicita MeSH
- retardanty hoření toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- organofosfáty * MeSH
- retardanty hoření MeSH
- triphenyl phosphate MeSH Prohlížeč
Herbicides and their metabolites are often detected in water bodies where they may cause adverse effects to non-target organisms. Their effects at environmentally relevant concentrations are often unclear, especially concerning mixtures of pesticides. This study thus investigated the impacts of one of the most used herbicides: S-metolachlor and its two metabolites, metolachlor oxanilic acid (MOA) and metolachlor ethanesulfonic acid (MESA) on the development of zebrafish embryos (Danio rerio). Embryos were exposed to the individual substances and their environmentally relevant mixture until 120 hpf (hours post-fertilization). The focus was set on sublethal endpoints such as malformations, hatching success, length of fish larvae, spontaneous movements, heart rate and locomotion. Moreover, expression levels of eight genes linked to the thyroid system disruption, oxidative stress defense, mitochondrial metabolism, regulation of cell cycle and retinoic acid (RA) signaling pathway were analyzed. Exposure to S-metolachlor (1 μg/L) and the pesticide mixture (1 μg/L of each substance) significantly reduced spontaneous tail movements of 21 hpf embryos. Few rare developmental malformations were observed, but only in larvae exposed to more than 100 μg/L of individual substances (craniofacial deformation, non-inflated gas bladder, yolk sac malabsorption) and to 30 μg/L of each substance in the pesticide mixture (spine deformation). No effect on hatching success, length of larvae, heart rate or larvae locomotion were found. Strong responses were detected at the molecular level including induction of p53 gene regulating the cell cycle (the pesticide mixture - 1 μg/L of each substance; MESA 30 μg/L; and MOA 100 μg/L), as induction of cyp26a1 gene encoding cytochrome P450 (pesticide mixture - 1 μg/L of each substance). Genes implicated in the thyroid system regulation (dio2, thra, thrb) were all overexpressed by the environmentally relevant concentrations of the pesticide mixture (1 μg/L of each substance) and MESA metabolite (1 μg/L). Zebrafish thyroid system disruption was revealed by the overexpressed genes, as well as by some related developmental malformations (mainly gas bladder and yolk sac abnormalities), and reduced spontaneous tail movements. Thus, the thyroid system disruption represents a likely hypothesis behind the effects caused by the low environmental concentrations of S-metolachlor, its two metabolites and their mixture.
- Klíčová slova
- Embryotoxicity, Environmental concentration, Metabolite, Pesticide mixture, Sublethal effects, Zebrafish embryo,
- MeSH
- acetamidy metabolismus toxicita MeSH
- chemické látky znečišťující vodu metabolismus toxicita MeSH
- dánio pruhované metabolismus MeSH
- embryo nesavčí účinky léků metabolismus MeSH
- embryonální vývoj účinky léků MeSH
- herbicidy metabolismus toxicita MeSH
- larva MeSH
- štítná žláza účinky léků embryologie MeSH
- synergismus léků MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- acetamidy MeSH
- chemické látky znečišťující vodu MeSH
- herbicidy MeSH
- metolachlor MeSH Prohlížeč
Schizophrenia is a severe disorder characterized by positive, negative and cognitive symptoms, which are still not fully understood. The development of efficient antipsychotics requires animal models of a strong validity, therefore the aims of the article were to summarize the construct, face and predictive validity of schizophrenia models based on rodents and zebrafish, to compare the advantages and disadvantages of these models, and to propose future directions in schizophrenia modeling and indicate when it is reasonable to combine these models. The advantages of rodent models stem primarily from the high homology between rodent and human physiology, neurochemistry, brain morphology and circuitry. The advantages of zebrafish models stem in the high fecundity, fast development and transparency of the embryo. Disadvantages of both models originate in behavioral repertoires not allowing specific symptoms to be modeled, even when the models are combined. Especially modeling the verbal component of certain positive, negative and cognitive symptoms is currently impossible.
- Klíčová slova
- animal models, laboratory rodents, model validity, neurobiology, schizophrenia, schizophrenia symptoms, zebrafish,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Mycobacterium tuberculosis, the etiologic agent of tuberculosis, is an intracellular pathogen of alveolar macrophages. These cells avidly take up nanoparticles, even without the use of specific targeting ligands, making the use of nanotherapeutics ideal for the treatment of such infections. Methoxy poly(ethylene oxide)- block-poly(ε-caprolactone) nanoparticles of several different polymer blocks' molecular weights and sizes (20-110 nm) were developed and critically compared as carriers for rifampicin, a cornerstone in tuberculosis therapy. The polymeric nanoparticles' uptake, consequent organelle targeting and intracellular degradation were shown to be highly dependent on the nanoparticles' physicochemical properties (the cell uptake half-lives 2.4-21 min, the degradation half-lives 51.6 min-ca. 20 h after the internalization). We show that the nanoparticles are efficiently taken up by macrophages and are able to effectively neutralize the persisting bacilli. Finally, we demonstrate, using a zebrafish model of tuberculosis, that the nanoparticles are well tolerated, have a curative effect, and are significantly more efficient compared to a free form of rifampicin. Hence, these findings demonstrate that this system shows great promise, both in vitro and in vivo, for the treatment of tuberculosis.
- MeSH
- dánio pruhované MeSH
- lidé MeSH
- makrofágy * metabolismus mikrobiologie MeSH
- modely nemocí na zvířatech MeSH
- Mycobacterium tuberculosis růst a vývoj MeSH
- myši MeSH
- nanočástice * chemie terapeutické užití MeSH
- nosiče léků * chemie farmakokinetika farmakologie MeSH
- RAW 264.7 buňky MeSH
- rifampin * chemie farmakokinetika farmakologie MeSH
- tuberkulóza farmakoterapie metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- nosiče léků * MeSH
- rifampin * MeSH
Hematopoiesis is a precisely orchestrated process regulated by the activity of hematopoietic cytokines and their respective receptors. Due to an extra round of whole genome duplication during vertebrate evolution in teleost fish, zebrafish have two paralogs of many important genes, including genes involved in hematopoiesis. Importantly, these duplication events brought increased level of complexity in such cases, where both ligands and receptors have been duplicated in parallel. Therefore, precise understanding of binding specificities between duplicated ligand-receptor signalosomes as well as understanding of their differential expression provide an important basis for future studies to better understand the role of duplication of these genes. However, although many recent studies in the field have partly addressed functional redundancy or sub-specialization of some of those duplicated paralogs, this information remains to be scattered over many publications and unpublished data. Therefore, the focus of this review is to provide an overview of recent findings in the zebrafish hematopoietic field regarding activity, role and specificity of some of the hematopoietic cytokines with emphasis on crucial regulators of the erythro-myeloid lineages.
- Klíčová slova
- cytokine, erythropoiesis, genome duplication, hematopoiesis, myelopoiesis, zebrafish,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Cyanobacteria are known for their ability to produce and release mixtures of up to thousands of compounds into the environment. Recently, the production of novel metabolites, retinoids, was reported for some cyanobacterial species along with teratogenic effects of samples containing these compounds. Retinoids are natural endogenous substances derived from vitamin A that play a crucial role in early vertebrate development. Disruption of retinoid signalling- especially during the early development of the nervous system- might lead to major malfunctions and malformations. In this study, the toxicity of cyanobacterial biomass samples from the field containing retinoids was characterized by in vivo and in vitro bioassays with a focus on the potential hazards towards nervous system development and function. Additionally, in order to identify the compounds responsible for the observed in vitro and in vivo effects the complex cyanobacterial extracts were fractionated (C18 column, water-methanol gradient) and the twelve obtained fractions were tested in bioassays. In all bioassays, all-trans retinoic acid (ATRA) was tested along with the environmental samples as a positive control. Retinoid-like activity (mediated via the retinoic acid receptor, RAR) was measured in the transgenic cell line p19/A15. The in vitro assay showed retinoid-like activity by specific interaction with RAR for the biomass samples. Neurotoxic effects of selected samples were studied on zebrafish (Danio rerio) embryos using the light/dark transition test (Viewpoint, ZebraLab system) with 120 hpf larvae. In the behavioural assay, the cyanobacterial extracts caused significant hyperactivity in zebrafish at 120 hpf after acute exposure (3 h prior to the measurement) at concentrations below the teratogenicity LOEC (0.2 g dw L-1). Similar effect was observed after exposure to fractions of the extracts with detected retinoid-like activity and additive effect was observed after combining the fractions. However, the effect on behaviour was not observed after exposure to ATRA only. To provide additional insight into the behavioural effects and describe the underlying mechanism gene expression of selected biomarkers was measured. We evaluated an array of 28 genes related to general toxicity, neurodevelopment, retinoid and thyroid signalling. We detected several affected genes, most notably, the Cyp26 enzymes that control endogenous ATRA concentration, which documents an effect on retinoid signalling.
- Klíčová slova
- ATRA, Biomarkers, Cyanobacteria, Locomotor response, Retinoids, Zebrafish,
- MeSH
- biomasa MeSH
- biotest MeSH
- chemické látky znečišťující vodu metabolismus toxicita MeSH
- chování zvířat účinky léků MeSH
- dánio pruhované růst a vývoj metabolismus MeSH
- embryo nesavčí účinky léků metabolismus MeSH
- exprese genu účinky léků MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- receptory kyseliny retinové genetika metabolismus MeSH
- sinice růst a vývoj metabolismus MeSH
- tretinoin metabolismus toxicita MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chemické látky znečišťující vodu MeSH
- receptory kyseliny retinové MeSH
- tretinoin MeSH
The aim of the study was to investigate the effects of subchronic exposure of zebrafish (Danio rerio) to a fluoroquinolone norfloxacin, using selected oxidative stress parameters as a target. Toxicity tests were performed on zebrafish according to the OECD Guidelines number 203 and number 215. In the Subchronic Toxicity Test, a significant (P < 0.01) increase in the activity of glutathione peroxidase, glutathione S-transferase, and catalase was found. In the test, norfloxacin did not affect lipid peroxidation and catalytic activity of glutathione reductase. From the results, we can conclude that norfloxacin has a negative impact on specific biochemical processes connected with the production of reactive oxygen species in fish tested.
- MeSH
- antibakteriální látky škodlivé účinky farmakologie MeSH
- dánio pruhované metabolismus MeSH
- norfloxacin škodlivé účinky farmakologie MeSH
- oxidační stres účinky léků MeSH
- reaktivní formy kyslíku metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibakteriální látky MeSH
- norfloxacin MeSH
- reaktivní formy kyslíku MeSH
Multiple molecular targets have been identified to mediate membrane-delimited and nongenomic effects of natural and synthetic steroids, but the influence of steroid metabolism on neuroactive steroid signaling is not well understood. To begin to address this question, we set out to identify major metabolites of a neuroprotective synthetic steroid 20-oxo-5β-pregnan-3α-yl l-glutamyl 1-ester (pregnanolone glutamate, PAG) and characterize their effects on GABAA and NMDA receptors (GABARs, NMDARs) and their influence on zebrafish behavior. Gas chromatography-mass spectrometry was used to assess concentrations of PAG and its metabolites in the hippocampal tissue of juvenile rats following intraperitoneal PAG injection. PAG is metabolized in the peripheral organs and nervous tissue to 20-oxo-17α-hydroxy-5β-pregnan-3α-yl l-glutamyl 1-ester (17-hydroxypregnanolone glutamate, 17-OH-PAG), 3α-hydroxy-5β-pregnan-20-one (pregnanolone, PA), and 3α,17α-dihydroxy-5β-pregnan-20-one (17-hydroxypregnanolone, 17-OH-PA). Patch-clamp electrophysiology experiments in cultured hippocampal neurons demonstrate that PA and 17-OH-PA are potent positive modulators of GABARs, while PAG and 17-OH-PA have a moderate inhibitory effect at NMDARs. PAG, 17-OH-PA, and PA diminished the locomotor activity of zebrafish larvae in a dose-dependent manner. Our results show that PAG and its metabolites are potent modulators of neurotransmitter receptors with behavioral consequences and indicate that neurosteroid-based ligands may have therapeutic potential.
- Klíčová slova
- glutamate, negative allosteric modulator, steroid, thigmotaxis, zebrafish,
- MeSH
- dánio pruhované MeSH
- estery MeSH
- GABA MeSH
- krysa rodu Rattus MeSH
- kyselina glutamová MeSH
- pregnanolon * farmakologie chemie MeSH
- receptory GABA-A MeSH
- receptory N-methyl-D-aspartátu * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- estery MeSH
- GABA MeSH
- kyselina glutamová MeSH
- pregnanolon * MeSH
- receptory GABA-A MeSH
- receptory N-methyl-D-aspartátu * MeSH
Zebrafish (Danio rerio) is a commonly-used vertebrate model species for many research areas. However, its low milt volume limits effective cryopreservation of sperm from a single individual and often precludes dividing a single semen sample to conduct multiple downstream procedures such as genomic DNA/RNA extraction and in-vitro fertilization. Here, we apply germ stem cell transplantation to increase zebrafish sperm production in a closely related larger species from the same subfamily, giant danio Devario aequipinnatus. The endogenous germ cell of the host is depleted by dead-end morpholino antisense oligonucleotide. Histology of the sterile gonad and quantitative PCR of gonadal tissue reveals all sterile giant danio develop the male phenotype. Spermatogonial cells of Tg(ddx4:egfp) transgenic zebrafish are transplanted into sterile giant danio larvae, and 22% of recipients (germline chimera) produce donor-derived sperm at sexual maturation. The germline chimera produce approximately three-fold the volume of sperm and 10-fold the spermatozoon concentration of the donor. The donor-derived sperm is functional and gives rise to viable progeny upon fertilization of donor oocytes. We show that the issue of low milt volume can be effectively addressed by employing a larger surrogate parent.
- MeSH
- Cyprinidae * MeSH
- dánio pruhované * genetika MeSH
- sperma MeSH
- spermatogonie MeSH
- spermie MeSH
- transplantace kmenových buněk MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of this study was to assess the impact of sulfamethoxazole (SMX) on oxidative stress indices in zebrafish (Danio rerio). The test was completed after 14 days. The tested concentrations were 50, 100 and 500 µg/L of SMX. Glutathione peroxidase, glutathione reductase, glutathione S-transferase and lipid peroxidation were investigated to determine the effects of SMX on oxidative stress in zebrafish. Lipid peroxidation gradually increased slightly (but non-significantly) at all tested concentrations during the test as compared to the control. The evaluation of oxidative stress biomarkers showed no significant changes in the activity of antioxidant enzymes in any experimental group exposed to SMX as compared to the control. The gradual increase in lipid peroxidation after 3 and 14 days in the SMX treated groups as compared to the control group indicates increasing cell membrane damage.
- Klíčová slova
- Fish, TBARS, antibiotics, antioxidant enzymes, oxidative stress,
- MeSH
- antibakteriální látky toxicita MeSH
- antioxidancia metabolismus MeSH
- časové faktory MeSH
- dánio pruhované metabolismus MeSH
- oxidační stres účinky léků MeSH
- peroxidace lipidů účinky léků MeSH
- sulfamethoxazol toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- antioxidancia MeSH
- sulfamethoxazol MeSH
The physiology of males and females can be vastly different, complicating interpretation of toxicological and physiological data. The objectives of this study were to elucidate the sex differences in the microbiome-gastrointestinal (GI) transcriptome of adult zebrafish. We compared microbial composition and diversity in both males and females fed the same diet and housed in the same environment. There were no sex-specific differences in weight gain nor gastrointestinal morphology based on histopathology. There was no difference in gut microbial diversity, richness (Shannon and Chao1 index) nor predicted functional composition of the microbiome between males and females. Prior to post-hoc correction, male zebrafish showed higher abundance for the bacterial families Erythrobacteraceae and Lamiaceae, both belonging to the phyla Actinobacteria and Proteobacteria. At the genus level, Lamia and Altererythrobacter were more dominant in males and an unidentified genus in Bacteroidetes was more abundant in females. There were 16 unique differentially expressed transcripts in the gastrointestinal tissue between male and female zebrafish (FDR corrected, p < 0.05). Relative to males, the mRNA expression for trim35-9, slc25a48, chchd3b, csad, and hsd17b3 were lower in female GI while cyp2k6, adra2c, and bckdk were higher in the female GI. Immune and lipid-related gene network expression differed between the sexes (i.e., cholesterol export and metabolism) as well as networks related to gastric motility, gastrointestinal system absorption and digestion. Such data provide clues as to putative differences in gastrointestinal physiology between male and female zebrafish. This study identifies host-transcriptome differences that can be considered when interpreting the microgenderome of zebrafish in studies investigating GI physiology and toxicology of fishes.
- Klíčová slova
- Experimental design, Gastrointestinal system, Gene networks, Microgenderome, Sex differences,
- MeSH
- Bacteria MeSH
- dánio pruhované genetika MeSH
- gastrointestinální trakt mikrobiologie MeSH
- mikrobiota * MeSH
- proteiny regulující apoptózu MeSH
- střevní mikroflóra * genetika MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- proteiny regulující apoptózu MeSH
Benzotriazole ultraviolet stabilizers (BUVSs) are widely used additives in industrial materials and personal care products that protect products from ultraviolet damage. Due to their high production volume and potential to bioaccumulate, BUVSs are an environmental pollutant of concern. In this study, juvenile zebrafish (Danio rerio) were exposed to 4 BUVSs (UV-234, UV-326, UV-329, and UV-P) at 10 and 100 μg/L for 28 d. BUVSs induced hepatic vacuolization and nuclei pyknosis in the liver following 100 μg/L UV-234 and UV-329 exposure. Transcriptomic analysis in the liver uncovered pathways related to inflammation that were affected by BUVSs. Based upon these data, we measured the expression levels of 9 genes involved in AHR-IL17/IL22 pathway in zebrafish larvae exposed to each BUVSs at one dose of either 10 or 100 μg/L for 6 days in a second set experiment. Transcript levels of interleukins il17a and il22 were decreased, while il6 mRNA was increased with exposure to UV-234, UV-329, and UV-P. No change to targeted transcripts was observed with UV-326 treatments. Moreover, cyp1a1 and ahr2 levels were increased in larvae treated with 100 μg/L UV-329 or UV-P. Consistent with expression data, protein abundance of IL22 was decreased by 29% with exposure to 100 μg/L UV-P. Taken together, these results demonstrate that exposure to different benzotriazole congeners may be associated with immunotoxicity in zebrafish through the AHR-IL17/IL22 pathway, and this may be associated with hepatic damage with prolonged exposures. This study provides new insight into unique pathways perturbed by specific BUVSs congeners.
- Klíčová slova
- AHR-IL17/IL22 pathway, Benzotriazole ultraviolet stabilizers, Hepatic damage, Immunotoxicology, Liver transcriptome,
- MeSH
- dánio pruhované * MeSH
- larva MeSH
- triazoly MeSH
- ultrafialové záření * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- benzotriazole MeSH Prohlížeč
- triazoly MeSH
OBJECTIVES: We tested the toxicity of ethinylestradiol, a semisynthetic estrogen used in oral contraceptives, on all-male triploid zebrafish using commercial feeds and three different doses concentrations. We aimed to determine whether ethinylestradiol peroral administration resulted in vitellogenin production and whether all-male triploid zebrafish could serve as a model species for xenoestrogen testing. METHODS: The actual concentrations of 17α-ethinylestradiol were 0.0035 (low); 0.0315 (medium) and 0.365 (high) µg/g. Positive control represented commercial feeds containing 0.0465 µg/g of β-estradiol. The experiment lasted 8 weeks. RESULTS: Our results indicate that 17α-ethinylestradiol consumption does induce vitellogenin production in triploid zebrafish. CONCLUSIONS: The simple presence of vitellogenin is a definite symptom indicative of the potential for such changes due to the action of estrogenic substances. As such, this experiment has shown that the use of all-male triploid zebrafish populations, rather than the mixed-sex populations of other species previously used, could serve as a suitable alternative model population for controlled testing of the effects of xenoestrogens on fish.
- MeSH
- chemické látky znečišťující vodu * MeSH
- dánio pruhované * genetika MeSH
- estrogeny farmakologie MeSH
- ethinylestradiol toxicita MeSH
- triploidie MeSH
- vitelogeniny genetika MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chemické látky znečišťující vodu * MeSH
- estrogeny MeSH
- ethinylestradiol MeSH
- vitelogeniny MeSH
In this work, we carried out neurochemical and behavioral analysis of zebrafish (Danio rerio) treated with rotenone, an agent used to chemically induce a syndrome resembling Parkinson's disease (PD). Dopamine release, measured with fast-scan cyclic voltammetry (FSCV) at carbon-fiber electrodes in acutely harvested whole brains, was about 30% of that found in controls. Uptake, represented by the first order rate constant (k) and the half-life (t1/2) determined by nonlinear regression modeling of the stimulated release plots, was also diminished. Behavioral analysis revealed that rotenone treatment increased the time required for zebrafish to reach a reward within a maze by more than 50% and caused fish to select the wrong pathway, suggesting that latent learning was impaired. Additionally, zebrafish treated with rotenone suffered from diminished locomotor activity, swimming shorter distances with lower mean velocity and acceleration. Thus, the neurochemical and behavioral approaches, as applied, were able to resolve rotenone-induced differences in key parameters. This approach may be effective for screening therapies in this and other models of neurodegeneration.
- MeSH
- dánio pruhované metabolismus MeSH
- dopamin metabolismus MeSH
- kognice MeSH
- modely nemocí na zvířatech MeSH
- Parkinsonova nemoc * MeSH
- rotenon * farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- dopamin MeSH
- rotenon * MeSH