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Zebrafish 3.0: A Model for Toxicological Research
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Zebrafish 3.0: A Model for Toxicological Research

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Toxicology".

Deadline for manuscript submissions: closed (31 October 2021) | Viewed by 35300

Special Issue Editor

Dr. Luca Del Giacco

E-Mail Website
Guest Editor
Dipartimento di Bioscienze, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy
Interests: zebrafish; neurodevelopment/neurodegeneration (Parkinson’s disease, amyotrophic lateral sclerosis); ear development/hearing loss; ciliogenesis and ciliopathies; acute myeloid leukemia; zebrafish embryo xenografts; ecotoxicology
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Special Issue Information

Dear Colleagues,

The zebrafish (Danio rerio) has long been used as a model species for developmental biology studies. Zebrafish, however, offer considerably more as a model species and are now involved in many different research fields including, neurobiology, immunology, cancer, just to name a few. In addition, the employment of zebrafish embryos has enormously facilitated different sort of chemical testing, from pollutants toxicity assay to high-throughput drug screening. The relevant genetic homology with humans, the applicability of genome editing and transgenesis technologies, together with easy and low-cost protocols to expose the embryos to different compounds, make zebrafish a powerful tool for modeling human diseases and better understanding cellular and molecular mechanisms of toxicants and new therapeutics. Indeed, the potential of zebrafish is becoming increasingly evident, and the availability of these models allows for increased understanding of the role of chemical exposure in human disease. In this Special Issue, we welcome contributions that show recent developments in zebrafish toxicity testing and drug discovery, such as novel endpoints, assays and testing strategies. Articles covering aspects of cellular and molecular mechanisms of action to effects on individual fish as well as multigenerational effects and population level impacts are invited. Contributions on high-throughput screening, monitoring and safety assessment of chemicals are invited, as well as new insights on the effects in chemical exposure in zebrafish models of human diseases. Translational studies that employ molecular methods to enhance our understanding of human health or environmental risk assessment are also of interest.

Dr. Luca Del Giacco
Guest Editor

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Keywords

  • Toxicology
  • Ecotoxicology
  • Pharmacology
  • Toxicity testing
  • Developmental biology
  • Human health
  • Disease genetics
  • Epigenetics
  • Zebrafish
  • Vertebrate
  • Hazard assessment
  • Risk assessment
  • Molecular mechanisms
  • High throughput screening
  • Multigenerational
  • Systems biology
  • Translational study

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Published Papers (6 papers)

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Research

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12 pages, 2217 KiB  
Article
Inner Ear and Muscle Developmental Defects in Smpx-Deficient Zebrafish Embryos
by Anna Ghilardi, Alberto Diana, Renato Bacchetta, Nadia Santo, Miriam Ascagni, Laura Prosperi and Luca Del Giacco
Int. J. Mol. Sci. 2021, 22(12), 6497; https://doi.org/10.3390/ijms22126497 - 17 Jun 2021
Cited by 5 | Viewed by 3065
Abstract
The last decade has witnessed the identification of several families affected by hereditary non-syndromic hearing loss (NSHL) caused by mutations in the SMPX gene and the loss of function has been suggested as the underlying mechanism. In the attempt to confirm this hypothesis [...] Read more.
The last decade has witnessed the identification of several families affected by hereditary non-syndromic hearing loss (NSHL) caused by mutations in the SMPX gene and the loss of function has been suggested as the underlying mechanism. In the attempt to confirm this hypothesis we generated an Smpx-deficient zebrafish model, pointing out its crucial role in proper inner ear development. Indeed, a marked decrease in the number of kinocilia together with structural alterations of the stereocilia and the kinocilium itself in the hair cells of the inner ear were observed. We also report the impairment of the mechanotransduction by the hair cells, making SMPX a potential key player in the construction of the machinery necessary for sound detection. This wealth of evidence provides the first possible explanation for hearing loss in SMPX-mutated patients. Additionally, we observed a clear muscular phenotype consisting of the defective organization and functioning of muscle fibers, strongly suggesting a potential role for the protein in the development of muscle fibers. This piece of evidence highlights the need for more in-depth analyses in search for possible correlations between SMPX mutations and muscular disorders in humans, thus potentially turning this non-syndromic hearing loss-associated gene into the genetic cause of dysfunctions characterized by more than one symptom, making SMPX a novel syndromic gene. Full article
(This article belongs to the Special Issue Zebrafish 3.0: A Model for Toxicological Research)
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9 pages, 6523 KiB  
Article
Toxicity of Jegosaponins A and B from Styrax japonica Siebold et al. Zuccarini in Prostate Cancer Cells and Zebrafish Embryos Resulting from Increased Membrane Permeability
by Moe Nishimura, Hiroyuki Fuchino, Kaoru Takayanagi, Hitomi Kawakami, Hiroko Nakayama, Nobuo Kawahara and Yasuhito Shimada
Int. J. Mol. Sci. 2021, 22(12), 6354; https://doi.org/10.3390/ijms22126354 - 14 Jun 2021
Cited by 10 | Viewed by 2976
Abstract
(1) Background: Screening of medicinal herbs is one of the most powerful approaches to identifying novel therapeutic molecules against many human diseases. To avoid potential harmful effects during medicinal use, toxicity testing is necessary in the early stages of drug discovery. The objective [...] Read more.
(1) Background: Screening of medicinal herbs is one of the most powerful approaches to identifying novel therapeutic molecules against many human diseases. To avoid potential harmful effects during medicinal use, toxicity testing is necessary in the early stages of drug discovery. The objective of this study was to identify the cytotoxic mechanisms of jegosaponin A and B from Styrax japonica Siebold et al. Zuccarini; (2) Methods: We screened Japanese medicinal herb extracts using PC-3 prostate cancer cells and found that a methanol extract isolated from the unripe fruit of Styrax japonica Siebold et al. Zuccarini (SJSZ) had an inhibitory effect on cell viability. We further performed fractionation assays with PC-3 cells and identified the bioactive compounds using LC/MS and NMR analysis. We clarified the toxic mechanisms of these compounds using PC-3 cells and zebrafish embryos; (3) Results: We identified two active molecules, jegosaponin A and jegosaponin B, in the inhibitory fractions of the methanol extract. These jegosaponins are toxic to zebrafish embryos during the early developmental stage. Jegosaponin A and B showed strong haemolytic activity in sheep defibrinated blood (EC50 = 2.1 μM, and 20.2 μM, respectively) and increased the cell membrane permeability in PC-3 cells and zebrafish embryos, which were identified using a membrane non-permeable DRAQ7, a fluorescent nucleus staining dye; (4) We identified the cytotoxic compounds jegosaponin A and B from SJSZ, which we showed to exhibit cell membrane disruptive properties using cell- and zebrafish-based testing. Full article
(This article belongs to the Special Issue Zebrafish 3.0: A Model for Toxicological Research)
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11 pages, 1826 KiB  
Article
Acute Toxicity of Cu-MOF Nanoparticles (nanoHKUST-1) towards Embryos and Adult Zebrafish
by Natalia Abramenko, Gregory Deyko, Evgeny Abkhalimov, Vera Isaeva, Lyubov Pelgunova, Eugeny Krysanov and Leonid Kustov
Int. J. Mol. Sci. 2021, 22(11), 5568; https://doi.org/10.3390/ijms22115568 - 25 May 2021
Cited by 31 | Viewed by 4524
Abstract
Metal-organic frameworks (MOFs) demonstrate unique properties, which are prospective for drug delivery, catalysis, and gas separation, but their biomedical applications might be limited due to their obscure interactions with the environment and humans. It is important to understand their toxic effect on nature [...] Read more.
Metal-organic frameworks (MOFs) demonstrate unique properties, which are prospective for drug delivery, catalysis, and gas separation, but their biomedical applications might be limited due to their obscure interactions with the environment and humans. It is important to understand their toxic effect on nature before their wide practical application. In this study, HKUST-1 nanoparticles (Cu-nanoMOF, Cu3(btc)2, btc = benzene-1,3,5-tricarboxylate) were synthesized by the microwave (MW)-assisted ionothermal method and characterized by X-ray powder diffraction (XRD) and transmission electron microscopy (TEM) techniques. The embryotoxicity and acute toxicity of HKUST-1 towards embryos and adult zebrafish were investigated. To gain a better understanding of the effects of Cu-MOF particles towards Danio rerio (D. rerio) embryos were exposed to HKUST-1 nanoparticles (NPs) and Cu2+ ions (CuSO4). Cu2+ ions showed a higher toxic effect towards fish compared with Cu-MOF NPs for D. rerio. Both forms of fish were sensitive to the presence of HKUST-1 NPs. Estimated LC50 values were 2.132 mg/L and 1.500 mg/L for zebrafish embryos and adults, respectively. During 96 h of exposure, the release of copper ions in a stock solution and accumulation of copper after 96 h were measured in the internal organs of adult fishes. Uptake examination of the major internal organs did not show any concentration dependency. An increase in the number of copper ions in the test medium was found on the first day of exposure. Toxicity was largely restricted to copper release from HKUST-1 nanomaterials structure into solution. Full article
(This article belongs to the Special Issue Zebrafish 3.0: A Model for Toxicological Research)
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15 pages, 5974 KiB  
Article
Use of Zebrafish Embryo Assay to Evaluate Toxicity and Safety of Bioreactor-Grown Exopolysaccharides and Endopolysaccharides from European Ganoderma applanatum Mycelium for Future Aquaculture Applications
by Wan Abd Al Qadr Imad Wan-Mohtar, Zul Ilham, Adi Ainurzaman Jamaludin and Neil Rowan
Int. J. Mol. Sci. 2021, 22(4), 1675; https://doi.org/10.3390/ijms22041675 - 7 Feb 2021
Cited by 30 | Viewed by 7716
Abstract
Natural mycelial exopolysaccharide (EPS) and endopolysaccharide (ENS) extracted from bioreactor-cultivated European Ganoderma applanatum mushrooms are of potential high commercial value for both food and adjacent biopharmaceutical industries. In order to evaluate their potential toxicity for aquaculture application, both EPS (0.01–10 mg/mL) and ENS [...] Read more.
Natural mycelial exopolysaccharide (EPS) and endopolysaccharide (ENS) extracted from bioreactor-cultivated European Ganoderma applanatum mushrooms are of potential high commercial value for both food and adjacent biopharmaceutical industries. In order to evaluate their potential toxicity for aquaculture application, both EPS (0.01–10 mg/mL) and ENS (0.01–10 mg/mL) extracts were tested for Zebrafish Embryo Toxicity (ZFET); early development effects on Zebrafish Embryos (ZE) were also analyzed between 24 and 120 h post-fertilization (HPF). Both EPS and ENS are considered non-toxic with LC50 of 1.41 mg/mL and 0.87 mg/mL respectively. Both EPS and ENS did not delay hatching and teratogenic defect towards ZE with <1.0 mg/mL, respectively. No significant changes in the ZE heart rate were detected following treatment with the two compounds tested (EPS: 0.01–10 mg/mL: 176.44 ± 0.77 beats/min and ENS: 0.01–10 mg/mL: 148.44 ± 17.75 beats/min) compared to normal ZE (120–180 beats/min). These initial findings support future pre-clinical trials in adult fish models with view to safely using EPS and ENS as potential feed supplements for supplements for development of the aquaculture industry. Full article
(This article belongs to the Special Issue Zebrafish 3.0: A Model for Toxicological Research)
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Review

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23 pages, 917 KiB  
Review
Zebrafish Embryos and Larvae as Alternative Animal Models for Toxicity Testing
by Benedikt Bauer, Angela Mally and Daniel Liedtke
Int. J. Mol. Sci. 2021, 22(24), 13417; https://doi.org/10.3390/ijms222413417 - 14 Dec 2021
Cited by 64 | Viewed by 11222
Abstract
Prerequisite to any biological laboratory assay employing living animals is consideration about its necessity, feasibility, ethics and the potential harm caused during an experiment. The imperative of these thoughts has led to the formulation of the 3R-principle, which today is a pivotal scientific [...] Read more.
Prerequisite to any biological laboratory assay employing living animals is consideration about its necessity, feasibility, ethics and the potential harm caused during an experiment. The imperative of these thoughts has led to the formulation of the 3R-principle, which today is a pivotal scientific standard of animal experimentation worldwide. The rising amount of laboratory investigations utilizing living animals throughout the last decades, either for regulatory concerns or for basic science, demands the development of alternative methods in accordance with 3R to help reduce experiments in mammals. This demand has resulted in investigation of additional vertebrate species displaying favourable biological properties. One prominent species among these is the zebrafish (Danio rerio), as these small laboratory ray-finned fish are well established in science today and feature outstanding biological characteristics. In this review, we highlight the advantages and general prerequisites of zebrafish embryos and larvae before free-feeding stages for toxicological testing, with a particular focus on cardio-, neuro, hepato- and nephrotoxicity. Furthermore, we discuss toxicokinetics, current advances in utilizing zebrafish for organ toxicity testing and highlight how advanced laboratory methods (such as automation, advanced imaging and genetic techniques) can refine future toxicological studies in this species. Full article
(This article belongs to the Special Issue Zebrafish 3.0: A Model for Toxicological Research)
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22 pages, 805 KiB  
Review
Zebrafish as a Model for the Study of Lipid-Lowering Drug-Induced Myopathies
by Magda Dubińska-Magiera, Marta Migocka-Patrzałek, Damian Lewandowski, Małgorzata Daczewska and Krzysztof Jagla
Int. J. Mol. Sci. 2021, 22(11), 5654; https://doi.org/10.3390/ijms22115654 - 26 May 2021
Cited by 11 | Viewed by 4349
Abstract
Drug-induced myopathies are classified as acquired myopathies caused by exogenous factors. These pathological conditions develop in patients without muscle disease and are triggered by a variety of medicaments, including lipid-lowering drugs (LLDs) such as statins, fibrates, and ezetimibe. Here we summarise the current [...] Read more.
Drug-induced myopathies are classified as acquired myopathies caused by exogenous factors. These pathological conditions develop in patients without muscle disease and are triggered by a variety of medicaments, including lipid-lowering drugs (LLDs) such as statins, fibrates, and ezetimibe. Here we summarise the current knowledge gained via studies conducted using various models, such as cell lines and mammalian models, and compare them with the results obtained in zebrafish (Danio rerio) studies. Zebrafish have proven to be an excellent research tool for studying dyslipidaemias as a model of these pathological conditions. This system enables in-vivo characterization of drug and gene candidates to further the understanding of disease aetiology and develop new therapeutic strategies. Our review also considers important environmental issues arising from the indiscriminate use of LLDs worldwide. The widespread use and importance of drugs such as statins and fibrates justify the need for the meticulous study of their mechanism of action and the side effects they cause. Full article
(This article belongs to the Special Issue Zebrafish 3.0: A Model for Toxicological Research)
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