Övgüm Çolakoğlu^A., Nida Dereli^E.

Introduction

Model organisms can be utilized to comprehend particular biological processes and gather data that can bring light to how other organisms function. Many fish models have been used in the scientific world. Each fish species has specific benefits and drawbacks¹. For instance, research on growth, stress, immunology, and reproduction has been done using goldfish. The most common species of fish utilized to research genetics, reproduction, and development was the medaka fish. Recently, zebrafish have become more often used as a model organism².

The tropical freshwater fish Danio rerio, popularly known as zebrafish, is most commonly seen in Southeast Asia. In order to prevent attacks from predators, zebrafish are typically located on the bottom of slowly flowing water³. Zebrafish are considered omnivores due to their wide-ranging diets and omnivorous eating behaviors⁴.

Since zebrafish were less complex than mice and thus straightforward for genetic modifications, George Streisinger utilized it as a model organism for the first time in the 1970s⁵. Chuck Kimmel, a colleague of Streisinger, found the use of zebrafish embryos more interesting than the study of the nervous system. The 1990s saw the development of two significant genetic mutations in zebrafish, one by Nobel Prize winner Christiane Nusslein-Volhard and the other by Wolfgang Driever and Mark Fishman. This gave rise to the use of zebrafish as a model organism².

The Utility of Zebrafish as a Model Organism in Scientific Research

Zebrafish have gained popularity as an animal model in many research in recent years. It is widely used in core fields of study, including immunology, cardiology, physiology, nutrition, genetics, neuroscience, and developmental biology. The Zebrafish model has numerous inherent benefits over other species models⁶. This animal’s widespread use may be due to its inexpensive and simple laboratory maintenance. It is far simpler to sustain them in a lab setting than to recreate the circumstances necessary for mammals because of how simple their natural environment is. Their quick generation periods of three to six months accelerate the pace of experimentation⁷.

About 70% of human genes have at least one clear zebrafish orthologue, according to comparisons with the human reference genome⁸. That makes zebrafish suitable for understanding genetic problems and for genetic modifications. For instance, to research specific disorders, even rare ones, mutations can be made to zebrafish genes of interest and can be monitored⁹. Zebrafish embryos are transparent, and their entire growth process takes place outside of the mother’s body, making it an appealing organism¹⁰ (Figure 1)¹¹. Because of this special developmental process, researchers may examine every aspect of development, from beginning to adulthood¹.

Figure 1. A 24-hour-old zebrafish embryo and its morphology¹¹. It is easy to notice the anatomical structure since the embryo is transparent.

On the other hand, zebrafish have certain disadvantages. Due to certain organ differences, it is challenging to utilize zebrafish as a model for human reproduction or respiration. Another restriction is the inability to test some water-soluble medications in zebrafish because of their habitat¹.

The Usage Areas of the Zebrafish Model

1. Cardiovascular Diseases

Zebrafish have been used in several studies to examine the impact of mutations on the electric function of the vertebrate heart because their heart rates are more comparable to those of humans than those of other model species¹². When an adult zebrafish’s electrocardiography (ECG) is analyzed, it shows distinct P, QRS, and T waves with a QT length that suggests a similar repolarization period (Figure 2)¹³. Additionally, the zebrafish ventricular action potential (AP) resembles the human cardiac AP in that it has a prolonged plateau phase and, as a result, a unique QT interval on the ECG. Despite the fact that zebrafish and humans share many similarities, there are a few minor variances in AP dynamics that should be taken into consideration¹⁴.

Figure 2. Analyzing the electrocardiograms (ECG) of humans and zebrafish¹³. Zebrafish adult at age 23. (top) Human male 43 years old with good health (bottom). The same time scale is used to compare the recordings of the ECGs from zebrafish and humans directly.

Furthermore, zebrafish hearts can continue to regenerate for the rest of their lives. The astonishing capacity of zebrafish to regenerate their hearts after damage is one of the most compelling reasons to utilize them to study human cardiac regeneration¹⁵. The pathological process that the human heart goes through after a Myocardial infarction (MI), also known as heart attack, was accurately mimicked by a cryoinjury zebrafish model developed in 2011. Although a collagen scar develops between 14 and 21 days after cryoinjury, the zebrafish heart can remove the scar concurrently with myocardial regeneration, something that mammals cannot do. In zebrafish, as a result of an injury to the heart tissue, cell death, inflammatory responses, and high mechanical stresses come together, leading to the transformation of cells called fibroblasts into myofibroblasts and the production of structural components called collagen and extracellular matrix (ECM) in the wound area¹⁶. These structural components play an important role in maintaining the integrity of the heart wall after the death of heart cells. Interestingly, enzymes called matrix metalloproteinases (MMPs) are released by cells that regulate the inflammatory response, and heart cells break down the components of the ECM over time and dissolve it. MMPs have a significant role in cardiac remodeling and end-stage heart failure¹⁷. Because of that, zebrafish is useful in understanding the development of heart failure after MI and thus preventing it by creating novel therapeutic targets that might block certain MMP activities^16,18.

2. Metabolic Diseases

Zebrafish is a promising animal to investigate metabolic problems since it has several metabolic organs like humans¹. This fish shares the liver, intestine, pancreas, and kidney with humans as well as other metabolic organs¹⁹ (Figure 3)²⁰. In this small and simple model, it is possible to test the levels of insulin, blood glucose, lipids, and cholesterol using a variety of techniques, much like in mammals. To accurately imitate the real diseases of diabetes and obesity, various organizations have created models of metabolic illnesses¹⁹.

Figure 3. Human and zebrafish common organs²⁰. Humans (A), larval (B), and adult (C) zebrafish, together with their corresponding organs and tissues.

Adipocyte hypertrophy and hyperplasia are two main characteristics of obesity. Adult adipocytes have a single big lipid droplet, similar to human white adipose tissue²¹. Lipids are also accumulated in visceral, intramuscular, and subcutaneous adipocytes in zebrafish, offering the chance to comprehend how body fat distribution is regulated in conditions like obesity²². Oka and colleagues published the first diet-induced obese zebrafish model in 2010. For eight weeks, they overfeed the fish. A comparative transcriptome investigation has revealed that the lipid metabolism of the visceral adipose tissue of zebrafish, rats, mice, and humans is identical²³. In order to comprehend the disease in the context of systematic obesity, similar experiments and a variety of additional techniques for producing obesity in zebrafish were used, mirroring the most typical process occurring in persons affected by this condition^22,24.

3. Cancer

The zebrafish model has become a crucial tool for studying human cancer because of the similarity of the genomes of humans and zebrafish. The translucent embryos of zebrafish make it possible to observe the activity of cancer cells, particularly during metastasis, and this provides information on the processes behind the spread of cancer^25,26. Cancer also progresses more quickly in embryos, with tumor growth beginning two days after introduction. However, because adults’ organs are fully formed, they provide a more accurate in vivo model²⁷.

The use of the zebrafish model allows for the study of several molecular pathways for both the initiation and development of human cancer. Several different studies have been performed to generate leukemia, sarcoma, melanoma, neuroblastoma, and germ-cell tumor zebrafish models²⁸ (Figure 4)²⁹. Zebrafish cancer models can be created using a variety of techniques. Making mutant and transgenic lines is one approach. With this approach, it is possible to carefully track the impact of individual genes on the development of tumors²⁶. Another option is to implant human tumor cells into zebrafish, which allows for the concurrent study of many different tumor forms^25,30.

Figure 4. Genetically engineered zebrafish models to mimic human cancers²⁹. The size of the text varies according to how many zebrafish models of that particular cancer kind have been published. MPNST: malignant peripheral nerve sheath tumor.

A new model for cancer research termed xenograft assay involves implanting tumors into zebrafish³¹. Xenograft zebrafish models include introducing tagged cancer cells into several zebrafish embryonic locations to monitor their development, behavior, and interactions with the host’s microenvironment²⁷.

In 2005, Lee et al. showed the significance of zebrafish as a biological tool for cancer research and provided the first evidence of the compatibility of human cells and embryos. They carried out the first xenotransplantation test in zebrafish using dedifferentiated human melanoma cells³². The injection of these cells into zebrafish embryos at the blastula stage was observed throughout time. These findings supported the hypothesis that zebrafish embryos provide the cells with the signals required to selectively integrate into organs^32,33. To provide better, more precise outcomes that are more medically and biologically meaningful, several researchers have enhanced and refined the process, created new strategies, and carried out xenotransplantation using various cell lines³³. For example, researchers revealed that the zebrafish cxcr4b gene regulates neutrophil quantity and movement and that neutrophils collaborate with cancer cells to trigger early metastatic processes³¹. To test this, they introduced MDA-MB-231-B breast tumor cells into the bloodstream of both wild-type zebrafish embryos and zebrafish embryos lacking the cxcr4b gene. The researchers discovered that tumor cells in zebrafish embryos lacking the cxcr4b gene had no effect on neutrophil speed or distance traveled compared to wild-type embryos^31,34.

4. Neurological Diseases

The zebrafish is becoming more well-known as a model organism for translational neuroscience research. When comparing zebrafish brain structure to humans, there are differences, but the overall organization is similar³⁵ (Figure 5)³⁶. The zebrafish brain exhibits a fundamental resemblance with human neuroanatomical and neurochemical pathways. Key areas like the hypothalamus and olfactory bulb are present, and significant neurotransmitter systems like cholinergic, dopaminergic, and noradrenergic pathways are mapped in the brain. Zebrafish have a developing blood-brain barrier that becomes functional around 10 days after fertilization^36,37.

Figure 5. Human and larval zebrafish neuroanatomy comparison³⁶. Sagittal portions of adult human brains (top) and 5-day post-fertilization zebrafish brains (bottom). Despite the enormous contrast in dimension and complexity, homologous sections are colored the same colors to demonstrate the retention of basic vertebrate brain structure from human to larval zebrafish.

The zebrafish has gained widespread acceptance as a model for researching and treating neurological conditions including, schizophrenia, Huntington’s disease (HD), Parkinson’s disease (PD), and Alzheimer’s disease (AD)³⁷. Zebrafish share many of the same genes related to PD and are responsive to drugs associated with PD risk. This has allowed researchers to create various PD models in zebrafish, both genetically and chemically³⁸. Even though zebrafish do not have the same type of dopamine-producing neurons as humans in their midbrain, they do have a similar cluster of dopamine-producing cells in a different part of their brain. Additionally, their serotonin and histamine systems are quite similar to those in mammals³⁹. While unable to recapitulate the full spectrum of the disease, they can be particularly useful for studying movement problems seen in PD. For instance, by disrupting dopamine-producing cells in zebrafish, researchers can mimic one of the main symptoms of PD, which is slow movement (bradykinesia)⁴⁰.

Astroglia in zebrafish are frequently referred to as radial glia rather than astroglia since they resemble mammalian radial glia in many ways. In the process of developing the brain, radial glial cells perform crucial functions in structuring the brain and directing the proliferation of neurons^41,42. Dysfunction of radial glia may cause neurodevelopmental problems. Researchers discover more about the operation of radial glia and how they can be related to neurological disorders by examining them in zebrafish. The studies on adult zebrafish brains have already produced significant new possibilities that potentially increase the proliferation and neurogenesis of mammalian glial cells⁴².

In one of the most recent studies, the researchers studied the impact of Nerve growth factor receptor (Ngfr) signaling on neurogenesis and reactive gliosis in the brain using zebrafish as a model organism. They created a zebrafish adult model of AD and discovered that Ngfr signaling is sufficient to lower Lcn2 levels (a marker upregulated in several brain diseases) and promote a pro-neurogenic state by suppressing the molecular signatures of reactive astrogliosis under AD pathology. Additionally, scientists investigated what would occur if they inhibited the Lcn2 pathway’s Slc22a17, a protein that plays a role in nerve cell growth, and discovered that doing so increased the supporting activity for nerve cell growth. These findings imply that Ngfr signaling may be utilized in AD to improve brain function and reduce reactive astroglia signaling⁴³.

Drug Development and Toxicity

Zebrafish play a crucial role in drug screening due to their alignment with the 3Rs principles: replacement, reduction, and refinement⁴⁴. Certain animal-based toxicity studies can be replaced by zebrafish experiments, particularly those that use their larvae. Zebrafish larvae can be used as the main model for discovering toxic medication candidates. This reduces hazards by ensuring that safer compounds are investigated in mammalian models. The design of animal experiments is improved by using zebrafish embryos and larvae. These embryos are externally fertilized and transparent in the earliest stages, allowing for easy monitoring of harmful effects and potential recovery^44,45. Zebrafish respond to small molecules and drugs within physiological doses. When combined with gene editing technologies and specific reporters, drug activity can be studied at a single-cell level in a whole organism, spanning tissues and timeframes⁴⁶.

Zebrafish embryos are employed as a predictive model to evaluate toxicity in mammals. Comparing the lethal concentration (LC50) of different chemicals in zebrafish embryos and mammals reveals that zebrafish generally have a lower median lethal dose. Moreover, studies on organ toxicity have shown similarities between zebrafish and mammals^47,48. Important drugs have been evaluated using the zebrafish model⁴⁶. For instance, a method was developed to detect small molecule-induced mutagenesis in susceptible zebrafish. The model was also employed to compare developmental toxicity caused by exposure to substances like ethanol or acetaldehyde².

Conclusion

The zebrafish (Danio rerio) has emerged as a powerful and versatile model organism in the realm of scientific research. Its unique features and properties have made it prominent in studies covering a variety of fields, including cardiovascular disease, metabolic disorders, cancer, and neurological conditions^2,3. The zebrafish’s transparent embryos, rapid growth, simple maintenance requirements, and genetic similarity to humans have made it an ideal subject for studying^7,10. Its applicability extends from genetics and developmental biology to disease modeling and drug screening, offering insights that can potentially transform our understanding of biological processes and human health^19,26,37. As scientists continue to delve deeper into the complexities of life, the zebrafish stands as a valuable model, aiding researchers in uncovering fundamental insights that clear the way for innovative treatments and solutions across a broad spectrum of disciplines.

References:

1. Teame, T., Zhang, Z., Ran, C., Zhang, H., Yang, Y., Ding, Q., Xie, M., Gao, C., Ye, Y., Duan, M., & Zhou, Z. (2019). The use of zebrafish (Danio rerio) as biomedical models. Animal Frontiers, 9(3), 68–77. https://doi.org/10.1093/af/vfz020
2. Rahman Khan, F., & Sulaiman Alhewairini, S. (2019). Zebrafish ( Danio rerio) as a Model Organism. In Current Trends in Cancer Management. IntechOpen. https://doi.org/10.5772/intechopen.81517
3. Kandasamy, T., Chandrasekar, S., Pichaivel, M., Pachaiappan, S., Muthusamy, G., & Sumathi, L. (2022). A Review of Zebrafish as an Alternative Animal Model and Its Benefits over Other Animal Models in Various Disease Conditions. Saudi Journal of Biomedical Research, 7(12), 355–359. https://doi.org/10.36348/sjbr.2022.v07i12.005
4. Watts, S. A., & D’Abramo, L. R. (2021). Standardized Reference Diets for Zebrafish: Addressing Nutritional Control in Experimental Methodology. Annual Review of Nutrition, 41(1), 511–527. https://doi.org/10.1146/annurev-nutr-120420-034809
5. Tonelli, F., Bek, J. W., Besio, R., De Clercq, A., Leoni, L., Salmon, P., Coucke, P. J., Willaert, A., & Forlino, A. (2020). Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders. Frontiers in Endocrinology, 11. https://doi.org/10.3389/fendo.2020.00489
6. Nowik, N., Podlasz, P., Jakimiuk, A., Kasica, N., Sienkiewicz, W., & Kaleczyc, J. (2015). Zebrafish: an animal model for research in veterinary medicine. Polish Journal of Veterinary Sciences, 18(3), 663–674. https://doi.org/10.1515/pjvs-2015-0086
7. Hoo, J. Y., Kumari, Y., Shaikh, M. F., Hue, S. M., & Goh, B. H. (2016). Zebrafish: A Versatile Animal Model for Fertility Research. BioMed Research International, 2016, 1–20. https://doi.org/10.1155/2016/9732780
8. Howe, K., Clark, M. D., Torroja, C. F., Torrance, J., Berthelot, C., Muffato, M., Collins, J. E., Humphray, S., McLaren, K., Matthews, L., McLaren, S., Sealy, I., Caccamo, M., Churcher, C., Scott, C., Barrett, J. C., Koch, R., Rauch, G.-J., White, S., … Stemple, D. L. (2013). The zebrafish reference genome sequence and its relationship to the human genome. Nature, 496(7446), 498–503. https://doi.org/10.1038/nature12111
9. Crouzier, L., Richard, E., Sourbron, J., Lagae, L., Maurice, T., & Delprat, B. (2021). Use of Zebrafish Models to Boost Research in Rare Genetic Diseases. International Journal of Molecular Sciences, 22(24), 13356. https://doi.org/10.3390/ijms222413356
10. Yang, L., Ho, N. Y., Alshut, R., Legradi, J., Weiss, C., Reischl, M., Mikut, R., Liebel, U., Müller, F., & Strähle, U. (2009). Zebrafish embryos as models for embryotoxic and teratological effects of chemicals. Reproductive Toxicology, 28(2), 245–253. https://doi.org/10.1016/j.reprotox.2009.04.013
11. von Hellfeld, R., Brotzmann, K., Baumann, L., Strecker, R., & Braunbeck, T. (2020). Adverse effects in the fish embryo acute toxicity (FET) test: a catalogue of unspecific morphological changes versus more specific effects in zebrafish (Danio rerio) embryos. Environmental Sciences Europe, 32(1), 122. https://doi.org/10.1186/s12302-020-00398-3
12. González-Rosa, J. M. (2022). Zebrafish Models of Cardiac Disease: From Fortuitous Mutants to Precision Medicine. Circulation Research, 130(12), 1803–1826. https://doi.org/10.1161/CIRCRESAHA.122.320396
13. Vornanen, M., & Hassinen, M. (2016). Zebrafish heart as a model for human cardiac electrophysiology. Channels, 10(2), 101–110. https://doi.org/10.1080/19336950.2015.1121335
14. Duong, T., Rose, R., Blazeski, A., Fine, N., Woods, C. E., Thole, J. F., Sotoodehnia, N., Soliman, E. Z., Tung, L., McCallion, A. S., & Arking, D. E. (2021). Development and optimization of an in vivo electrocardiogram recording method and analysis program for adult zebrafish. Disease Models & Mechanisms, 14(8). https://doi.org/10.1242/dmm.048827
15. Major, R. J., & Poss, K. D. (2007). Zebrafish heart regeneration as a model for cardiac tissue repair. Drug Discovery Today: Disease Models, 4(4), 219–225. https://doi.org/10.1016/j.ddmod.2007.09.002
16. Beffagna, G. (2019). Zebrafish as a Smart Model to Understand Regeneration After Heart Injury: How Fish Could Help Humans. Frontiers in Cardiovascular Medicine, 6. https://doi.org/10.3389/fcvm.2019.00107
17. VANHOUTTE, D., SCHELLINGS, M., PINTO, Y., & HEYMANS, S. (2006). Relevance of matrix metalloproteinases and their inhibitors after myocardial infarction: A temporal and spatial window. Cardiovascular Research, 69(3), 604–613. https://doi.org/10.1016/j.cardiores.2005.10.002
18. Ross Stewart, K. M., Walker, S. L., Baker, A. H., Riley, P. R., & Brittan, M. (2022). Hooked on heart regeneration: the zebrafish guide to recovery. Cardiovascular Research, 118(7), 1667–1679. https://doi.org/10.1093/cvr/cvab214
19. Ghaddar, B., & Diotel, N. (2022). Zebrafish: A New Promise to Study the Impact of Metabolic Disorders on the Brain. International Journal of Molecular Sciences, 23(10), 5372. https://doi.org/10.3390/ijms23105372
20. Wang, X., Copmans, D., & de Witte, P. A. M. (2021). Using Zebrafish as a Disease Model to Study Fibrotic Disease. International Journal of Molecular Sciences, 22(12), 6404. https://doi.org/10.3390/ijms22126404
21. Flynn, E. J., Trent, C. M., & Rawls, J. F. (2009). Ontogeny and nutritional control of adipogenesis in zebrafish (Danio rerio). Journal of Lipid Research, 50(8), 1641–1652. https://doi.org/10.1194/jlr.M800590-JLR200
22. do Carmo Rodrigues Virote, B., Rodrigues da Cunha Barreto Vianna, A., & David Solis Murgas, L. (2020). Zebrafish as an Experimental Model for the Study of Obesity. In Zebrafish in Biomedical Research. IntechOpen. https://doi.org/10.5772/intechopen.88576
23. Oka, T., Nishimura, Y., Zang, L., Hirano, M., Shimada, Y., Wang, Z., Umemoto, N., Kuroyanagi, J., Nishimura, N., & Tanaka, T. (2010). Diet-induced obesity in zebrafish shares common pathophysiological pathways with mammalian obesity. BMC Physiology, 10(1), 21. https://doi.org/10.1186/1472-6793-10-21
24. Zang, L., Maddison, L. A., & Chen, W. (2018). Zebrafish as a Model for Obesity and Diabetes. Frontiers in Cell and Developmental Biology, 6. https://doi.org/10.3389/fcell.2018.00091
25. Astell, K. R., & Sieger, D. (2020). Zebrafish In Vivo Models of Cancer and Metastasis. Cold Spring Harbor Perspectives in Medicine, 10(8), a037077. https://doi.org/10.1101/cshperspect.a037077
26. Hason, & Bartůněk. (2019). Zebrafish Models of Cancer—New Insights on Modeling Human Cancer in a Non-Mammalian Vertebrate. Genes, 10(11), 935. https://doi.org/10.3390/genes10110935
27. Letrado, P., de Miguel, I., Lamberto, I., Díez-Martínez, R., & Oyarzabal, J. (2018). Zebrafish: Speeding Up the Cancer Drug Discovery Process. Cancer Research, 78(21), 6048–6058. https://doi.org/10.1158/0008-5472.CAN-18-1029
28.  Casey, M. J., & Stewart, R. A. (2020). Pediatric Cancer Models in Zebrafish. Trends in Cancer, 6(5), 407–418. https://doi.org/10.1016/j.trecan.2020.02.006
29. McConnell, A. M., Noonan, H. R., & Zon, L. I. (2021). Reeling in the Zebrafish Cancer Models. Annual Review of Cancer Biology, 5(1), 331–350. https://doi.org/10.1146/annurev-cancerbio-051320-014135
30. Moshal, K. S., Ferri-Lagneau, K. F., Haider, J., Pardhanani, P., & Leung, T. (2011). Discriminating Different Cancer Cells Using a Zebrafish in Vivo Assay. Cancers, 3(4), 4102–4113. https://doi.org/10.3390/cancers3044102
31. Chen, X., Li, Y., Yao, T., & Jia, R. (2021). Benefits of Zebrafish Xenograft Models in Cancer Research. Frontiers in Cell and Developmental Biology, 9. https://doi.org/10.3389/fcell.2021.616551
32. Lee, L. M. J., Seftor, E. A., Bonde, G., Cornell, R. A., & Hendrix, M. J. C. (2005). The fate of human malignant melanoma cells transplanted into zebrafish embryos: Assessment of migration and cell division in the absence of tumor formation. Developmental Dynamics, 233(4), 1560–1570. https://doi.org/10.1002/dvdy.20471
33. Liu, Y., Wu, W., Cai, C., Zhang, H., Shen, H., & Han, Y. (2023). Patient-derived xenograft models in cancer therapy: technologies and applications. Signal Transduction and Targeted Therapy, 8(1), 160. https://doi.org/10.1038/s41392-023-01419-2
34. Tulotta, C., Stefanescu, C., Chen, Q., Torraca, V., Meijer, A. H., & Snaar-Jagalska, B. E. (2019). CXCR4 signaling regulates metastatic onset by controlling neutrophil motility and response to malignant cells. Scientific Reports, 9(1), 2399. https://doi.org/10.1038/s41598-019-38643-2
35. Kalueff, A. V., Stewart, A. M., & Gerlai, R. (2014). Zebrafish as an emerging model for studying complex brain disorders. Trends in Pharmacological Sciences, 35(2), 63–75. https://doi.org/10.1016/j.tips.2013.12.002
36. Burgess, H. A., & Burton, E. A. (2023). A Critical Review of Zebrafish Neurological Disease Models−1. The Premise: Neuroanatomical, Cellular and Genetic Homology and Experimental Tractability. Oxford Open Neuroscience, 2. https://doi.org/10.1093/oons/kvac018
37. Best, J. (2008). Zebrafish: An in vivo model for the study of neurological diseases. Neuropsychiatric Disease and Treatment, 567. https://doi.org/10.2147/NDT.S2056
38. Razali, K., Othman, N., Mohd Nasir, M. H., Doolaanea, A. A., Kumar, J., Ibrahim, W. N., Mohamed Ibrahim, N., & Mohamed, W. M. Y. (2021). The Promise of the Zebrafish Model for Parkinson’s Disease: Today’s Science and Tomorrow’s Treatment. Frontiers in Genetics, 12. https://doi.org/10.3389/fgene.2021.655550
39. Chia, K., Klingseisen, A., Sieger, D., & Priller, J. (2022). Zebrafish as a model organism for neurodegenerative disease. Frontiers in Molecular Neuroscience, 15. https://doi.org/10.3389/fnmol.2022.940484
40. Doyle, J. M., & Croll, R. P. (2022). A Critical Review of Zebrafish Models of Parkinson’s Disease. Frontiers in Pharmacology, 13. https://doi.org/10.3389/fphar.2022.835827
41. Miranda-Negrón, Y., & García-Arrarás, J. E. (2022). Radial glia and radial glia-like cells: Their role in neurogenesis and regeneration. Frontiers in Neuroscience, 16. https://doi.org/10.3389/fnins.2022.1006037
42. Jurisch‐Yaksi, N., Yaksi, E., & Kizil, C. (2020). Radial glia in the zebrafish brain: Functional, structural, and physiological comparison with the mammalian glia. Glia, 68(12), 2451–2470. https://doi.org/10.1002/glia.23849
43. Siddiqui, T., Cosacak, M. I., Popova, S., Bhattarai, P., Yilmaz, E., Lee, A. J., Min, Y., Wang, X., Allen, M., İş, Ö., Atasavum, Z. T., Rodriguez-Muela, N., Vardarajan, B. N., Flaherty, D., Teich, A. F., Santa-Maria, I., Freudenberg, U., Werner, C., Tosto, G., … Kizil, C. (2023). Nerve growth factor receptor (Ngfr) induces neurogenic plasticity by suppressing reactive astroglial Lcn2/Slc22a17 signaling in Alzheimer’s disease. Npj Regenerative Medicine, 8(1), 33. https://doi.org/10.1038/s41536-023-00311-5
44. Cassar, S., Adatto, I., Freeman, J. L., Gamse, J. T., Iturria, I., Lawrence, C., Muriana, A., Peterson, R. T., Van Cruchten, S., & Zon, L. I. (2020). Use of Zebrafish in Drug Discovery Toxicology. Chemical Research in Toxicology, 33(1), 95–118. https://doi.org/10.1021/acs.chemrestox.9b00335
45. Modarresi Chahardehi, A., Arsad, H., & Lim, V. (2020). Zebrafish as a Successful Animal Model for Screening Toxicity of Medicinal Plants. Plants, 9(10), 1345. https://doi.org/10.3390/plants9101345
46. Patton, E. E., Zon, L. I., & Langenau, D. M. (2021). Zebrafish disease models in drug discovery: from preclinical modelling to clinical trials. Nature Reviews Drug Discovery, 20(8), 611–628. https://doi.org/10.1038/s41573-021-00210-8
47. Rosa, J. G. S., Lima, C., & Lopes-Ferreira, M. (2022). Zebrafish Larvae Behavior Models as a Tool for Drug Screenings and Pre-Clinical Trials: A Review. International Journal of Molecular Sciences, 23(12), 6647. https://doi.org/10.3390/ijms23126647
48. Ducharme, N. A., Reif, D. M., Gustafsson, J.-A., & Bondesson, M. (2015). Comparison of toxicity values across zebrafish early life stages and mammalian studies: Implications for chemical testing. Reproductive Toxicology, 55, 3–10. https://doi.org/10.1016/j.reprotox.2014.09.005