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