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Zebrafish Animal Models
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Zebrafish Animal Models

A special issue of Genes (ISSN 2073-4425). This special issue belongs to the section "Human Genomics and Genetic Diseases".

Deadline for manuscript submissions: closed (30 April 2021) | Viewed by 24707

Special Issue Editor

Prof. Dr. Laura Sánchez

E-Mail Website
Guest Editor
Department of Zoology, Genetics and Physical Anthropology, University of Santiago de Compostela, Santiago de Compostela, 15705 Galicia, Spain
Interests: zebrafish; Danio rerio; animal models for human disease; xenograft of tumor cells; toxicity
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The use of animal models has improved our understanding of many biological processes.

The importance of these models has grown in the last decade, especially those involving zebrafish (Danio rerio)—an emerging star in this group. Zebrafish were the last animal incorporated into the vertebrates, mainly due to their special characteristics. This species has a short generation time, its embryos are very robust and transparent, the number of descendants is very high, and maintenance is not expensive. Zebrafish have been extensively used in toxicological studies, drug screening, cancer research, and in the generation of animal models. Transgenesis and mutagenesis have been the most popular techniques for modeling, and with advances in genetics, the possibilities in this area have increased greatly. The use of CRISPR has created mutated lines that are being used as models for the study of rare and complex diseases. The comparison between morphants, obtained by the use of morpholines, and mutant phenotypes, generated by CRISPR, is contributing to our understanding of the function of genes related to diseases.

For this Special Issue in Genes, we invite the submission of both reviews and original research articles that use Zebrafish as a model in the study of human diseases.

Prof. Laura Sánchez
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Genes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • zebrafish
  • Danio rerio
  • animal models for human diseases
  • transgenesis
  • mutagenesis

Published Papers (7 papers)

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Research

Jump to: Review

21 pages, 6705 KiB  
Article
Conservation of Zebrafish MicroRNA-145 and Its Role during Neural Crest Cell Development
by Tomás J. Steeman, Juan A. Rubiolo, Laura E. Sánchez, Nora B. Calcaterra and Andrea M. J. Weiner
Genes 2021, 12(7), 1023; https://0-doi-org.brum.beds.ac.uk/10.3390/genes12071023 - 30 Jun 2021
Cited by 5 | Viewed by 2709
Abstract
The neural crest is a multipotent cell population that develops from the dorsal neural fold of vertebrate embryos in order to migrate extensively and differentiate into a variety of tissues. A number of gene regulatory networks coordinating neural crest cell specification and differentiation [...] Read more.
The neural crest is a multipotent cell population that develops from the dorsal neural fold of vertebrate embryos in order to migrate extensively and differentiate into a variety of tissues. A number of gene regulatory networks coordinating neural crest cell specification and differentiation have been extensively studied to date. Although several publications suggest a common role for microRNA-145 (miR-145) in molecular reprogramming for cell cycle regulation and/or cellular differentiation, little is known about its role during in vivo cranial neural crest development. By modifying miR-145 levels in zebrafish embryos, abnormal craniofacial development and aberrant pigmentation phenotypes were detected. By whole-mount in situ hybridization, changes in expression patterns of col2a1a and Sry-related HMG box (Sox) transcription factors sox9a and sox9b were observed in overexpressed miR-145 embryos. In agreement, zebrafish sox9b expression was downregulated by miR-145 overexpression. In silico and in vivo analysis of the sox9b 3′UTR revealed a conserved potential miR-145 binding site likely involved in its post-transcriptional regulation. Based on these findings, we speculate that miR-145 participates in the gene regulatory network governing zebrafish chondrocyte differentiation by controlling sox9b expression. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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12 pages, 1203 KiB  
Article
Morphological Abnormalities and Gene Expression Changes Caused by High Incubation Temperatures in Zebrafish Xenografts with Human Cancer Cells
by Pablo Cabezas-Sainz, Carlos Coppel, Alba Pensado-López, Pedro Fernandez, Laura Muinelo-Romay, Rafael López-López, Juan A. Rubiolo and Laura Sánchez
Genes 2021, 12(1), 113; https://0-doi-org.brum.beds.ac.uk/10.3390/genes12010113 - 19 Jan 2021
Cited by 4 | Viewed by 2573
Abstract
Published studies show that most of the human cancer xenograft studies in zebrafish embryos have used incubation temperatures in the range of 32–34 °C for 3–6 days post-injection, trying to find a compromise temperature between the zebrafish embryos (28 °C) and the human [...] Read more.
Published studies show that most of the human cancer xenograft studies in zebrafish embryos have used incubation temperatures in the range of 32–34 °C for 3–6 days post-injection, trying to find a compromise temperature between the zebrafish embryos (28 °C) and the human injected cells (37 °C). While this experimental setup is widely used, a question remains: is possible to overcome the drawbacks caused by a suboptimal temperature for the injected cells? To clarify the effect of temperature and injected cells on the host, in this study, we analyzed the development and health of the last in response to different temperatures in the presence or absence of injected human cancer cells. Comparing different incubation temperatures (28, 34 and 36 °C), we determined morphological abnormalities and developmental effects in injected and non-injected embryos at different time points. Besides this, the expression of selected genes was determined by qPCR to determine temperature affected metabolic processes in the embryos. The results indicate that an incubation temperature of 36 °C during a period of 48 h is suitable for xenotransplantation without morphological or metabolic changes that could be affecting the host or the injected cells, allowing them to proliferate near their optimal temperature. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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19 pages, 7345 KiB  
Article
Alexander Disease Modeling in Zebrafish: An In Vivo System Suitable to Perform Drug Screening
by Simona Candiani, Silvia Carestiato, Andreas F. Mack, Daniele Bani, Matteo Bozzo, Valentina Obino, Michela Ori, Francesca Rosamilia, Miriam De Sarlo, Mario Pestarino, Isabella Ceccherini and Tiziana Bachetti
Genes 2020, 11(12), 1490; https://0-doi-org.brum.beds.ac.uk/10.3390/genes11121490 - 11 Dec 2020
Cited by 5 | Viewed by 3598
Abstract
Alexander disease (AxD) is a rare astrogliopathy caused by heterozygous mutations, either inherited or arising de novo, on the glial fibrillary acid protein (GFAP) gene (17q21). Mutations in the GFAP gene make the protein prone to forming aggregates which, together with heat-shock protein [...] Read more.
Alexander disease (AxD) is a rare astrogliopathy caused by heterozygous mutations, either inherited or arising de novo, on the glial fibrillary acid protein (GFAP) gene (17q21). Mutations in the GFAP gene make the protein prone to forming aggregates which, together with heat-shock protein 27 (HSP27), αB-crystallin, ubiquitin, and proteasome, contribute to form Rosenthal fibers causing a toxic effect on the cell. Unfortunately, no pharmacological treatment is available yet, except for symptom reduction therapies, and patients undergo a progressive worsening of the disease. The aim of this study was the production of a zebrafish model for AxD, to have a system suitable for drug screening more complex than cell cultures. To this aim, embryos expressing the human GFAP gene carrying the most severe p.R239C under the control of the zebrafish gfap gene promoter underwent functional validation to assess several features already observed in in vitro and other in vivo models of AxD, such as the localization of mutant GFAP inclusions, the ultrastructural analysis of cells expressing mutant GFAP, the effects of treatments with ceftriaxone, and the heat shock response. Our results confirm that zebrafish is a suitable model both to study the molecular pathogenesis of GFAP mutations and to perform pharmacological screenings, likely useful for the search of therapies for AxD. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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14 pages, 1614 KiB  
Article
Rad21 Haploinsufficiency Prevents ALT-Associated Phenotypes in Zebrafish Brain Tumors
by Aurora Irene Idilli, Cecilia Pazzi, Francesca dal Pozzolo, Michela Roccuzzo and Maria Caterina Mione
Genes 2020, 11(12), 1442; https://0-doi-org.brum.beds.ac.uk/10.3390/genes11121442 - 30 Nov 2020
Cited by 2 | Viewed by 2622
Abstract
Cohesin is a protein complex consisting of four core subunits responsible for sister chromatid cohesion in mitosis and meiosis, and for 3D genome organization and gene expression through the establishment of long distance interactions regulating transcriptional activity in the interphase. Both roles are [...] Read more.
Cohesin is a protein complex consisting of four core subunits responsible for sister chromatid cohesion in mitosis and meiosis, and for 3D genome organization and gene expression through the establishment of long distance interactions regulating transcriptional activity in the interphase. Both roles are important for telomere integrity, but the role of cohesin in telomere maintenance mechanisms in highly replicating cancer cells in vivo is poorly studied. Here we used a zebrafish model of brain tumor, which uses alternative lengthening of telomeres (ALT) as primary telomere maintenance mechanism to test whether haploinsufficiency for Rad21, a member of the cohesin ring, affects ALT development. We found that a reduction in Rad21 levels prevents ALT-associated phenotypes in zebrafish brain tumors and triggers an increase in tert expression. Despite the rescue of ALT phenotypes, tumor cells in rad21+/− fish exhibit an increase in DNA damage foci, probably due to a reduction in double-strand breaks repair efficiency. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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32 pages, 6100 KiB  
Article
Duplicated dnmt3aa and dnmt3ab DNA Methyltransferase Genes Play Essential and Non-Overlapped Functions on Modulating Behavioral Control in Zebrafish
by Yu-Heng Lai, Gilbert Audira, Sung-Tzu Liang, Petrus Siregar, Michael Edbert Suryanto, Huan-Chau Lin, Omar Villalobos, Oliver B. Villaflores, Erwei Hao, Ken-Hong Lim and Chung-Der Hsiao
Genes 2020, 11(11), 1322; https://0-doi-org.brum.beds.ac.uk/10.3390/genes11111322 - 07 Nov 2020
Cited by 7 | Viewed by 3579
Abstract
DNA methylation plays several roles in regulating neuronal proliferation, differentiation, and physiological functions. The major de novo methyltransferase, DNMT3, controls the DNA methylation pattern in neurons according to environmental stimulations and behavioral regulations. Previous studies demonstrated that knockout of Dnmt3 induced mouse anxiety; [...] Read more.
DNA methylation plays several roles in regulating neuronal proliferation, differentiation, and physiological functions. The major de novo methyltransferase, DNMT3, controls the DNA methylation pattern in neurons according to environmental stimulations and behavioral regulations. Previous studies demonstrated that knockout of Dnmt3 induced mouse anxiety; however, controversial results showed that activation of Dnmt3 causes anxiolytic behavior. Thus, an alternative animal model to clarify Dnmt3 on modulating behavior is crucial. Therefore, we aimed to establish a zebrafish (Danio rerio) model to clarify the function of dnmt3 on fish behavior by behavioral endpoint analyses. We evaluated the behaviors of the wild type, dnmt3aa, and dnmt3ab knockout (KO) fish by the novel tank, mirror biting, predator avoidance, social interaction, shoaling, circadian rhythm locomotor activity, color preference, and short-term memory tests. The results indicated that the dnmt3aa KO fish possessed abnormal exploratory behaviors and less fear response to the predator. On the other hand, dnmt3ab KO fish displayed less aggression, fear response to the predator, and interests to interact with their conspecifics, loosen shoaling formation, and dysregulated color preference index ranking. Furthermore, both knockout fishes showed higher locomotion activity during the night cycle, which is a sign of anxiety. However, changes in some neurotransmitter levels were observed in the mutant fishes. Lastly, whole-genome DNA methylation sequencing demonstrates a potential network of Dnmt3a proteins that is responsive to behavioral alterations. To sum up, the results suggested that the dnmt3aa KO or dnmt3ab KO fish display anxiety symptoms, which supported the idea that Dnmt3 modulates the function involved in emotional control, social interaction, and cognition. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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9 pages, 1737 KiB  
Article
Danio Rerio as Model Organism for Adenoviral Vector Evaluation
by Paola Gulías, Jorge Guerra-Varela, Manuela Gonzalez-Aparicio, Ana Ricobaraza, Africa Vales, Gloria Gonzalez-Aseguinolaza, Rubén Hernandez-Alcoceba and Laura Sánchez
Genes 2019, 10(12), 1053; https://0-doi-org.brum.beds.ac.uk/10.3390/genes10121053 - 17 Dec 2019
Cited by 7 | Viewed by 3535
Abstract
Viral vector use is wide-spread in the field of gene therapy, with new clinical trials starting every year for different human pathologies and a growing number of agents being approved by regulatory agencies. However, preclinical testing is long and expensive, especially during the [...] Read more.
Viral vector use is wide-spread in the field of gene therapy, with new clinical trials starting every year for different human pathologies and a growing number of agents being approved by regulatory agencies. However, preclinical testing is long and expensive, especially during the early stages of development. Nowadays, the model organism par excellence is the mouse (Mus musculus), and there are few investigations in which alternative models are used. Here, we assess the possibility of using zebrafish (Danio rerio) as an in vivo model for adenoviral vectors. We describe how E1/E3-deleted adenoviral vectors achieve efficient transduction when they are administered to zebrafish embryos via intracranial injection. In addition, helper-dependent (high-capacity) adenoviral vectors allow sustained transgene expression in this organism. Taking into account the wide repertoire of genetically modified zebrafish lines, the ethical aspects, and the affordability of this model, we conclude that zebrafish could be an efficient alternative for the early-stage preclinical evaluation of adenoviral vectors. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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Review

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45 pages, 1600 KiB  
Review
Experimental Models to Study Autism Spectrum Disorders: hiPSCs, Rodents and Zebrafish
by Alba Pensado-López, Sara Veiga-Rúa, Ángel Carracedo, Catarina Allegue and Laura Sánchez
Genes 2020, 11(11), 1376; https://0-doi-org.brum.beds.ac.uk/10.3390/genes11111376 - 20 Nov 2020
Cited by 15 | Viewed by 5241
Abstract
Autism Spectrum Disorders (ASD) affect around 1.5% of the global population, which manifest alterations in communication and socialization, as well as repetitive behaviors or restricted interests. ASD is a complex disorder with known environmental and genetic contributors; however, ASD etiology is far from [...] Read more.
Autism Spectrum Disorders (ASD) affect around 1.5% of the global population, which manifest alterations in communication and socialization, as well as repetitive behaviors or restricted interests. ASD is a complex disorder with known environmental and genetic contributors; however, ASD etiology is far from being clear. In the past decades, many efforts have been put into developing new models to study ASD, both in vitro and in vivo. These models have a lot of potential to help to validate some of the previously associated risk factors to the development of the disorder, and to test new potential therapies that help to alleviate ASD symptoms. The present review is focused on the recent advances towards the generation of models for the study of ASD, which would be a useful tool to decipher the bases of the disorder, as well as to conduct drug screenings that hopefully lead to the identification of useful compounds to help patients deal with the symptoms of ASD. Full article
(This article belongs to the Special Issue Zebrafish Animal Models)
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