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The Zebrafish Model in Animal and Human Health Research

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: closed (30 September 2024) | Viewed by 13546

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Dr. Marta Anna Szychlinska

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Department of Biomedicine, Neuroscience and Advanced Diagnostics (BiND), Human Anatomy Section, University of Palermo, 90127 Palermo, Italy
Interests: articular cartilage engineering; chondrogenesis; mesenchymal stem cells; aging; hypertrophy; morphology; pathophysiology; osteoarthritis
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Dr. Antonella Marino Gammazza

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Department of Biomedicine, Neuroscience and Advanced Diagnostics Institute of Human Anatomy and Histology, University of Palermo, Palermo, Italy
Interests: anatomy; oxidative stress; molecular chaperones; cell signaling; cell biology; bioinformatics
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Special Issue Information

Dear Colleagues,

The zebrafish (Danio rerio) has become a widely used vertebrate model for biomedical and ecotoxicological research. Due to its high fertility rate, small size, rapid development and optical transparency during early development as well as its strong genetic similarity with humans, the zebrafish is particularly useful both to model human diseases and to address several questions about the effects of environmental contaminants on human development and health. Zebrafish are used as a model in all areas of environmental toxicology involving the study of the biological effects of toxic compounds in the natural environment, including their effects on humans and animals. Zebrafish are increasingly being employed in predictive toxicology, in the screening of contaminants and in the assessment of environmental matrices, permitting monitoring and, finally, allowing researchers to find adequate preventive strategies to protect life on planet Earth.

In this Special Issue of the International Journal of Molecular Sciences, we focus our attention on the latest discoveries, methods and technology advancements as well as environmental toxicant assessments in zebrafish models which can provide knowledge which can be used to monitor and preserve animal and human health. We aim to provide a comprehensive update regarding the literature accessible to scientists within this field. Thus, we wish to invite investigators from basic, clinical and translational biomedical and ecotoxicological research or closely related disciplines to contribute with original articles, reviews, communications and concept papers.

Dr. Marta Anna Szychlinska
Dr. Antonella Marino Gammazza
Guest Editors

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Keywords

zebrafish
environmental toxicology
predictive toxicology

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Published Papers (8 papers)

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Research

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22 pages, 6138 KiB

Open AccessArticle

Genetic and Environmental Factors Co-Contributing to Behavioral Abnormalities in adnp/adnp2 Mutant Zebrafish

by Yongxin Wang, Xiaoyun Sun, Bo Xiong, Ming Duan and Yuhua Sun

Int. J. Mol. Sci. 2024, 25(17), 9469; https://doi.org/10.3390/ijms25179469 - 30 Aug 2024

Viewed by 978

Abstract

Human mutations of ADNP and ADNP2 are known to be associated with neural developmental disorders (NDDs), including autism spectrum disorders (ASDs) and schizophrenia (SZ). However, the underlying mechanisms remain elusive. In this study, using CRISPR/Cas9 gene editing technology, we generated adnp and adnp2 mutant zebrafish models, which exhibited developmental delays, brain deficits, and core behavioral features of NDDs. RNA sequencing analysis of adnpa⁻^/⁻; adnpb^−/⁻ and adnp2a^−/⁻; adnp2b^−/⁻ larval brains revealed altered gene expression profiles affecting synaptic transmission, autophagy, apoptosis, microtubule dynamics, hormone signaling, and circadian rhythm regulation. Validation using whole-mount in situ hybridization (WISH) and real-time quantitative PCR (qRT-PCR) corroborated these findings, supporting the RNA-seq results. Additionally, loss of adnp and adnp2 resulted in significant downregulation of pan-neuronal HuC and neuronal fiber network α-Tubulin signals. Importantly, prolonged low-dose exposure to environmental endocrine disruptors (EEDs) aggravated behavioral abnormalities in adnp and adnp2 mutants. This comprehensive approach enhances our understanding of the complex interplay between genetic mutations and environmental factors in NDDs. Our findings provide novel insights and experimental foundations into the roles of adnp and adnp2 in neurodevelopment and behavioral regulation, offering a framework for future preclinical drug screening aimed at elucidating the pathogenesis of NDDs and related conditions. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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20 pages, 6906 KiB

Open AccessArticle

Altered Morpho-Functional Features of Neurogenesis in Zebrafish Embryos Exposed to Non-Combustion-Derived Magnetite

by Pietro Cacialli, Serena Ricci, Giulia Pia Servetto, Valeria Franceschini, Francisco Ruiz-Zepeda and Ruggero Vigliaturo

Int. J. Mol. Sci. 2024, 25(12), 6459; https://doi.org/10.3390/ijms25126459 - 12 Jun 2024

Viewed by 1043

Abstract

Neurogenesis is the process by which new brain cells are formed. This crucial event emerges during embryonic life and proceeds in adulthood, and it could be influenced by environmental pollution. Non-combustion-derived magnetite represents a portion of the coarse particulate matter (PM) contributing to air and water pollution in urban settings. Studies on humans have reported that magnetite and other iron oxides have significant damaging effects at a central level, where these particles accumulate and promote oxidative stress. Similarly, magnetite nanoparticles can cross the placenta and damage the embryo brain during development, but the impact on neurogenesis is still unknown. Furthermore, an abnormal Fe cation concentration in cells and tissues might promote reactive oxygen species (ROS) generation and has been associated with multiple neurodegenerative conditions. In the present study, we used zebrafish as an in vivo system to analyze the specific effects of magnetite on embryonic neurogenesis. First, we characterized magnetite using mineralogical and spectroscopic analyses. Embryos treated with magnetite at sub-lethal concentrations showed a dose–response increase in ROS in the brain, which was accompanied by a massive decrease in antioxidant genes (sod2, cat, gsr, and nrf2). In addition, a higher number of apoptotic cells was observed in embryos treated with magnetite. Next, interestingly, embryos exposed to magnetite displayed a decrease in neural staminal progenitors (nestin, sox2, and pcna markers) and a neuronal marker (elavl3). Finally, we observed significative increases in apoeb (specific microglia marker) and interleukin-1b (il1b), confirming a status of inflammation in the brain embryos treated with magnetite. Our study represents the very first in vivo evidence concerning the effects of magnetite on brain development. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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12 pages, 2812 KiB

Open AccessArticle

High NaCl Concentrations in Water Are Associated with Developmental Abnormalities and Altered Gene Expression in Zebrafish

by Denis A. Seli, Andrew Prendergast, Yagmur Ergun, Antariksh Tyagi and Hugh S. Taylor

Int. J. Mol. Sci. 2024, 25(7), 4104; https://doi.org/10.3390/ijms25074104 - 7 Apr 2024

Cited by 2 | Viewed by 1317

Abstract

Salt is frequently introduced in ecosystems, where it acts as a pollutant. This study examined how changes in salinity affect the survival and development of zebrafish from the two-cell to the blastocyst stage and from the blastocyst to the larval stage. Control zebrafish embryos were cultured in E3 medium containing 5 mM Sodium Chloride (NaCl), 0.17 mM Potassium Chloride (KCL), 0.33 mM Calcium Chloride (CaCl₂), and 0.33 mM Magnesium Sulfade (MgSO₄). Experiments were conducted using increasing concentrations of each individual salt at 5×, 10×, 50×, and 100× the concentration found in E3 medium. KCL, CaCl₂, and MgSO₄ did not result in lethal abnormalities and did not affect early embryo growth at any of the concentrations tested. Concentrations of 50× and 100× NaCl caused embryonic death in both stages of development. Concentrations of 5× and 10× NaCl resulted in uninflated swim bladders in 12% and 65% of larvae, compared to 4.2% of controls, and caused 1654 and 2628 genes to be differentially expressed in blastocysts, respectively. The ATM signaling pathway was affected, and the Sonic Hedgehog pathway genes Shh and Ptc1 implicated in swim bladder development were downregulated. Our findings suggest that increased NaCl concentrations may alter gene expression and cause developmental abnormalities in animals found in affected ecosystems. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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17 pages, 3164 KiB

Open AccessArticle

Antioxidant, Pro-Survival and Pro-Regenerative Effects of Conditioned Medium from Wharton’s Jelly Mesenchymal Stem Cells on Developing Zebrafish Embryos

by Chiara Reina, Clara Cardella, Margot Lo Pinto, Gaia Pucci, Santina Acuto, Aurelio Maggio and Vincenzo Cavalieri

Int. J. Mol. Sci. 2023, 24(17), 13191; https://doi.org/10.3390/ijms241713191 - 25 Aug 2023

Cited by 3 | Viewed by 1578

Abstract

Conditioned media harvested from stem cell culturing have the potential to be innovative therapeutic tools against various diseases, due to their high content of growth, trophic and protective factors. The evaluation in vivo of the effects and biosafety of these products is essential, and zebrafish provides an ideal platform for high-throughput toxicological analysis, concurrently allowing the minimization of the use of mammalian models without losing reliability. In this study, we assessed the biological effects elicited by the exposure of zebrafish embryos to a conditioned medium derived from Wharton’s jelly mesenchymal stem cells. By a multiparametric investigation combining molecular, embryological, behavioural and in vivo imaging techniques, we found that exposure to a conditioned medium at a non-toxic/non-lethal dosage triggers antioxidant, anti-apoptotic and pro-regenerative effects, by upregulation of a set of genes involved in antioxidant defence (nrf2, brg1, sirt1, sirt6, foxO3a, sod2 and cat), glycolysis (ldha) and cell survival (bcl2l1, mcl1a and bim), coupled to downregulation of pro-apoptotic markers (baxa, caspase-3a and caspase-8). To our knowledge, this is the first study comprehensively addressing the effects of a conditioned medium on a whole organism from a developmental, molecular and behavioural perspective, and we are fairly confident that it will pave the way for future therapeutic application. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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19 pages, 4451 KiB

Open AccessArticle

Transcriptome Analysis Reveals the Molecular Basis of Overfeeding-Induced Diabetes in Zebrafish

by Guodong Ge, Jing Ren, Guili Song, Qing Li and Zongbin Cui

Int. J. Mol. Sci. 2023, 24(15), 11994; https://doi.org/10.3390/ijms241511994 - 26 Jul 2023

Cited by 2 | Viewed by 1889

Abstract

Diabetes has gradually become a serious disease that threatens human health. It can induce various complications, and the pathogenesis of diabetes is quite complex and not yet fully elucidated. The zebrafish has been widely acknowledged as a useful model for investigating the mechanisms underlying the pathogenesis and therapeutic interventions of diabetes. However, the molecular basis of zebrafish diabetes induced by overfeeding remains unknown. In this study, a zebrafish diabetes model was established by overfeeding, and the molecular basis of zebrafish diabetes induced by overfeeding was explored. Compared with the control group, the body length, body weight, and condition factor index of zebrafish increased significantly after four weeks of overfeeding. There was a significant elevation in the fasting blood glucose level, accompanied by a large number of lipid droplets accumulated within the liver. The levels of triglycerides and cholesterol in both the serum and liver exhibited a statistically significant increase. Transcriptome sequencing was employed to investigate changes in the livers of overfed zebrafish. The number of up-regulated and down-regulated differentially expressed genes (DEGs) was 1582 and 2404, respectively, in the livers of overfed zebrafish. The DEGs were subjected to KEGG and GO enrichment analyses, and the hub signaling pathways and hub DEGs were identified. The results demonstrate that sixteen genes within the signal pathway associated with fatty acid metabolism were found to be significantly up-regulated. Specifically, these genes were found to mainly participate in fatty acid transport, fatty acid oxidation, and ketogenesis. Furthermore, thirteen genes that play a crucial role in glucose metabolism, particularly in the pathways of glycolysis and glycogenesis, were significantly down-regulated in the livers of overfed zebrafish. These results indicate insulin resistance and inhibition of glucose entry into liver cells in the livers of overfed zebrafish. These findings elucidate the underlying molecular basis of zebrafish diabetes induced by overfeeding and provide a model for further investigation of the pathogenesis and therapeutics of diabetes. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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18 pages, 4953 KiB

Open AccessArticle

Simple Coumarins from Peucedanum luxurians Fruits: Evaluation of Anxiolytic Activity and Influence on Gene Expression Related to Anxiety in Zebrafish Model

by Jarosław Widelski, Natalia Kasica, Monika Maciąg, Simon Vlad Luca, Barbara Budzyńska, Dafina Fondai, Piotr Podlasz and Krystyna Skalicka-Woźniak

Int. J. Mol. Sci. 2023, 24(10), 8693; https://doi.org/10.3390/ijms24108693 - 12 May 2023

Cited by 1 | Viewed by 1689

Abstract

Anxiety is one of the most common central nervous system disorders, affecting at least one-quarter of the worldwide population. The medications routinely used for the treatment of anxiety (mainly benzodiazepines) are a cause of addiction and are characterized by many undesirable side effects. Thus, there is an important and urgent need for screening and finding novel drug candidates that can be used in the prevention or treatment of anxiety. Simple coumarins usually do not show side effects, or these effects are much lower than in the case of synthetic drugs acting on the central nervous system (CNS). This study aimed to evaluate the anxiolytic activity of three simple coumarins from Peucedanum luxurians Tamamsch, namely officinalin, stenocarpin isobutyrate, and officinalin isobutyrate, in a 5 dpf larval zebrafish model. Moreover, the influence of the tested coumarins on the expression of genes involved in the neural activity (c-fos, bdnf) or dopaminergic (th1), serotoninergic (htr1Aa, htr1b, htr2b), GABA-ergic (gabarapa, gabarapb), enkephalinergic (penka, penkb), and galaninergic (galn) neurotransmission was assessed by quantitative PCR. All tested coumarins showed significant anxiolytic activity, with officinalin as the most potent compound. The presence of a free hydroxyl group at position C-7 and the lack of methoxy moiety at position C-8 might be key structural features responsible for the observed effects. In addition, officinalin and its isobutyrate upregulated the expression of genes involved in neurotransmission and decreased the expression of genes connected with neural activity. Therefore, the coumarins from P. luxurians might be considered as promising drug candidates for the therapy of anxiety and related disorders. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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Review

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21 pages, 1735 KiB

Open AccessReview

Speeding up Glioblastoma Cancer Research: Highlighting the Zebrafish Xenograft Model

by Giusi Alberti, Maria Denise Amico, Celeste Caruso Bavisotto, Francesca Rappa, Antonella Marino Gammazza, Fabio Bucchieri, Francesco Cappello, Federica Scalia and Marta Anna Szychlinska

Int. J. Mol. Sci. 2024, 25(10), 5394; https://doi.org/10.3390/ijms25105394 - 15 May 2024

Viewed by 1806

Abstract

Glioblastoma multiforme (GBM) is a very aggressive and lethal primary brain cancer in adults. The multifaceted nature of GBM pathogenesis, rising from complex interactions between cells and the tumor microenvironment (TME), has posed great treatment challenges. Despite significant scientific efforts, the prognosis for GBM remains very poor, even after intensive treatment with surgery, radiation, and chemotherapy. Efficient GBM management still requires the invention of innovative treatment strategies. There is a strong necessity to complete cancer in vitro studies and in vivo studies to properly evaluate the mechanisms of tumor progression within the complex TME. In recent years, the animal models used to study GBM tumors have evolved, achieving highly invasive GBM models able to provide key information on the molecular mechanisms of GBM onset. At present, the most commonly used animal models in GBM research are represented by mammalian models, such as mouse and canine ones. However, the latter present several limitations, such as high cost and time-consuming management, making them inappropriate for large-scale anticancer drug evaluation. In recent years, the zebrafish (Danio rerio) model has emerged as a valuable tool for studying GBM. It has shown great promise in preclinical studies due to numerous advantages, such as its small size, its ability to generate a large cohort of genetically identical offspring, and its rapid development, permitting more time- and cost-effective management and high-throughput drug screening when compared to mammalian models. Moreover, due to its transparent nature in early developmental stages and genetic and anatomical similarities with humans, it allows for translatable brain cancer research and related genetic screening and drug discovery. For this reason, the aim of the present review is to highlight the potential of relevant transgenic and xenograft zebrafish models and to compare them to the traditionally used animal models in GBM research. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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19 pages, 1484 KiB

Open AccessReview

A Short Review of the Toxicity of Dentifrices—Zebrafish Model as a Useful Tool in Ecotoxicological Studies

by Piotr Stachurski, Wojciech Świątkowski, Andrzej Ciszewski, Katarzyna Sarna-Boś and Agnieszka Michalak

Int. J. Mol. Sci. 2023, 24(18), 14339; https://doi.org/10.3390/ijms241814339 - 20 Sep 2023

Cited by 1 | Viewed by 1715

Abstract

This review aims to summarize the literature data regarding the effects of different toothpaste compounds in the zebrafish model. Danio rerio provides an insight into the mechanisms of the ecotoxicity of chemicals as well as an assessment of their fate in the environment to determine long-term environmental impact. The regular use of adequate toothpaste with safe active ingredients possessing anti-bacterial, anti-inflammatory, anti-oxidant, and regenerative properties is one of the most effective strategies for oral healthcare. In addition to water, a typical toothpaste consists of a variety of components, among which three are of predominant importance, i.e., abrasive substances, fluoride, and detergents. These ingredients provide healthy teeth, but their environmental impact on living organisms are often not well-known. Each of them can influence a higher level of organization: subcellular, cellular, tissue, organ, individual, and population. Therefore, it is very important that the properties of a chemical are detected before it is released into the environment to minimize damage. An important part of a chemical risk assessment is the estimation of the ecotoxicity of a compound. The zebrafish model has unique advantages in environmental ecotoxicity research and has been used to study vertebrate developmental biology. Among others, the advantages of this model include its external, visually accessible development, which allows for providing many experimental manipulations. The zebrafish has a significant genetic similarity with other vertebrates. Nevertheless, translating findings from zebrafish studies to human risk assessment requires careful consideration of these differences. Full article

(This article belongs to the Special Issue The Zebrafish Model in Animal and Human Health Research)

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