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1
Kiess, W., A. Körner, and K. Landgraf. "Der Zebrafisch als in vivo-Modellsystem für Adipositas und assoziierte Erkrankungen." Adipositas - Ursachen, Folgeerkrankungen, Therapie 12, no. 04 (December 2018): 198–203. http://dx.doi.org/10.1055/s-0038-1676678.
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ZusammenfassungViele Erkenntnisse über Mechanismen, die für die Entstehung von Adipositas, und den damit einhergehenden Adipositas-assoziierten Erkrankungen, relevant sind, sind mit Hilfe der Modellorganismen Maus oder Ratte erarbeitet worden. In den letzten Jahren hat sich der Zebrafisch als weiteres, sehr geeignetes in-vivo-Modellsystem etabliert, da er verschiedene Vorteile gegenüber Maus und Ratte und anderen Modellorganismen aufweist. Zudem sind wesentliche Aspekte der Regulation des Energiemetabolismus, welche bei der Entstehung von Adipositas und Adipositas-assoziierten Folgeerkrankungen beim Menschen eine Rolle spielen, im Zebrafisch konserviert. Dies beinhaltet unter anderem Mechanismen der zentralen Regulation des Sättigungsgefühls, der Fettzellentwicklung, der ernährungsbedingten Anhäufung von Fettgewebe sowie der Körperfettverteilung. Aufgrund dessen stellt der Zebrafisch ein geeignetes in vivo-Modellsystem für die Untersuchung von Prozessen dar, welche in die Entstehung von Adipositas und deren Folgeerkrankungen involviert sind.
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Westhoff, J. H., and B. Tönshoff. "Nephronneogenese durch renale Progenitorzellen im adulten Zebrafisch." Der Nephrologe 6, no. 5 (August 31, 2011): 435–36. http://dx.doi.org/10.1007/s11560-011-0553-3.
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Schredelseker, Johann, and Gerhard Krumschnabel. "Der Zebrafisch als vielseitiges Modellsystem. Vom Zierfisch zum Forschungsobjekt." Biologie in unserer Zeit 39, no. 6 (December 2009): 389–97. http://dx.doi.org/10.1002/biuz.200910406.
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Norton, William H. J., and Laure Bally-Cuif. "Die Zebrafisch fgfr1a-Mutante zeigt eine genetische Basis der Aggression." BIOspektrum 18, no. 1 (February 2012): 46–48. http://dx.doi.org/10.1007/s12268-012-0143-6.
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Jörgens, K., K. Schäker, and J. Kroll. "Der Modellorganismus Zebrafisch in der biomedizinischen Grundlagenforschung: Anwendungen und Perspektiven in der vaskulären Biologie und Medizin." DMW - Deutsche Medizinische Wochenschrift 136, no. 37 (September 6, 2011): 1865–68. http://dx.doi.org/10.1055/s-0031-1286357.
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Konantz, Martina, Martijn H. Brugman, In-Hyun Park, George Q. Daley, Christiane Nuesslein-Volhard, Christopher Baum, and Claudia Lengerke. "The Zebrafish Homologue of the Murine Ecotropic Viral Integration Site-1 (. Evi-1) gene Regulates Zebrafish Embryonic Blood Development." Blood 114, no. 22 (November 20, 2009): 1461. http://dx.doi.org/10.1182/blood.v114.22.1461.1461.
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Abstract Abstract 1461 Poster Board I-484 The ecotropic viral integration site-1 (Evi-1) locus was originally identified as a common site of retroviral integration in murine myeloid tumors and was later shown to be one of the most potent oncogenenes associated with murine and human myeloid leukemia. More recent data suggest involvement of Evi-1 in embryonic hematopoiesis (Goyama et al, Cell Stem Cell 2008; Yuasa et al, EMBO J, 2005), yet the precise role and molecular regulation of Evi-1 during blood development remains poorly understood. The zebrafish model offers powerful tools for genetic and embryonic studies. Here, we study zebrafish embryonic development and human pluripotent stem cells to understand how evi-1 modulates early hematopoietic development. Loss-of-function studies were performed in vivo by injecting Morpholino oligonucleotides in zebrafish zygotes to inhibit evi-1 pre-mRNA splicing. To control for off-target effects, two separate morpholinos were designed and injected. N=100 zebrafish were analysed pro experiment in each group. Inhibition of evi-1 was confirmed by quantitative PCR comparison in morpholino-injected and control embryos. Hematopoietic development was followed in both morphants and wild-type embryos by simple microscopy and in situ hybridizations using known hematopoeitic markers in order to investigate the developmental time-point in which evi-1 regulates blood development. evi-1 morpholino injected zebrafisch embryo showed severely reduced numbers of circulating blood cells, consistent with the phenotype observed in Evi-1−/− mice. Additionally, hemorrhages were observed, suggesting concomittant defects of the endothelial lineage in evi-1 deficient fish. In situ hybridization analysis on 11-12 somite stage embryos revealed strong reduction of myeloid embryonic hematopoiesis (measured by pu.1 expression in the anterior lateral plate mesoderm), while no change was observed in primitive erythroid progenitor cells (monitored by gata1 expression) or overall in blood and endothelial precursors in the posterior lateral plate mesoderm (as monitored by scl expression). Taken together, our studies demonstrate a strong impact of evi-1 on zebrafish blood development, confirming the results from Evi-1−/− mice. As gata1 expression and therefore erythroid precursor cells in the posterior blood islands are unaffected in evi-1 morphants, our results support the hypothesis that the reduction of primitive yolk-sac erythrocytes in mutant mice was caused from hemorrhages from pericardial effusions. Since erythroid and myeloid cells derive from a common precursor, but gata1 expression was unaffected in knock-down embryos, we anticipate that evi-1 plays a specific role in the myeloid lineage, as shown by abolished pu.1 expression in the anterior LPM. evi-1 therefore probably affects differentiation, survival or proliferation of myeloid cells. Previous reports in adult hematopoietic cells show that evi-1 can interact with both gata1 and pu.1. However, our data suggest that this is not the case during embryonic development, since gata1 expression remained unaltered in morpholino-injected embryos. Furthermore, data in mice suggest that Evi-1 may modulate embryonic hematopoiesis by affecting hematopoietic stem cell proliferation through regulation of Gata2. Currently ongoing experiments in our laboratories focus on characterization of genetic interactions between evi-1, gata2 and pu.1 during zebrafish blood development. Amongst other, gata2 and respectively pu.1 mRNA are co-injected in evi-1 morphants to analyse whether they can rescue the blood phenotype. Moreover, selected findings in zebrafish embryonic development will be verified in the human using using in vitro differentiating human induced pluripotent stem (iPS) cells. First expression data generated by real-time PCR analysis showed differential expression of EVI-1 in embryoid bodies generated from human iPS cells, confirming our hypothesis that EVI-1 has specific effects during human blood development. Disclosures No relevant conflicts of interest to declare.
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You-Jin, Jeon. "Antioxidant Effect of Fucoidan from Miyeokgui, Marine Alga in Zebrafish Model." Journal of Chitin and Chitosan 20, no. 2 (June 30, 2015): 123–30. http://dx.doi.org/10.17642/jcc.20.2.7.
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Jiang, Yanjun, Ye Rong, Ruixue Wu, Yi Wen, and Liu Hu. "A Zebrafish Thrombosis Model for Assessing Antiplatelet Drugs." International Journal of Pharma Medicine and Biological Sciences 9, no. 1 (January 2020): 38–42. http://dx.doi.org/10.18178/ijpmbs.9.1.38-42.
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Singh, Chandra Bhushan, and Badre Alam Ansari. "Bioassay of Profenofos and Cypermethrin on Zebrafish, Daniorerio." Scholars Academic Journal of Biosciences 4, no. 7 (July 2016): 578–82. http://dx.doi.org/10.21276/sajb.2016.4.7.5.
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Oh, Junyoung, Eun-Jin Park, Seongeun Kang, and Seungheon Lee. "Exposure to Dithiopyr Alters Swimming Performance Parameters in Zebrafish." Journal of Life Science 26, no. 2 (February 25, 2016): 181–89. http://dx.doi.org/10.5352/jls.2016.26.2.181.
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He, Junling, Yi Ding, Natalia Nowik, Charel Jager, Muhamed N. H. Eeza, A. Alia, Hans J. Baelde, and Herman P. Spaink. "Leptin deficiency affects glucose homeostasis and results in adiposity in zebrafish." Journal of Endocrinology 249, no. 2 (May 2021): 125–34. http://dx.doi.org/10.1530/joe-20-0437.
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Leptin is a hormone which functions in the regulation of energy homeostasis via suppression of appetite. In zebrafish, there are two paralogous genes encoding leptin, called lepa and lepb. In a gene expression study, we found that the lepb gene, not the lepa gene, was significantly downregulated under the state of insulin-resistance in zebrafish larvae, suggesting that the lepb plays a role in glucose homeostasis. In the current study, we characterised lepb-deficient (lepb−/−) adult zebrafish generated via a CRISPR-CAS9 gene editing approach by investigating whether the disruption of the lepb gene would result in the development of type 2 diabetes mellitus (T2DM) and diabetic complications. We observed that lepb−/− adult zebrafish had an increase in body weight, length and visceral fat accumulation, compared to age-matched control zebrafish. In addition, lepb−/− zebrafish had significantly higher blood glucose levels compared to control zebrafish. These data collectively indicate that lepb−/− adult zebrafish display the features of T2DM. Furthermore, we showed that lepb−/− adult zebrafish had glomerular hypertrophy and thickening of the glomerular basement membrane, compared to control zebrafish, suggesting that lepb−/− adult zebrafish develop early signs of diabetic nephropathy. In conclusion, our results demonstrate that lepb regulates glucose homeostasis and adiposity in zebrafish, and suggest that lepb−/− mutant zebrafish are a promising model to investigate the role of leptin in the development of T2DM and are an attractive model to perform mechanistic and therapeutic research in T2DM and its complications.
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Mwaffo, Violet, Peng Zhang, Sebastián Romero Cruz, and Maurizio Porfiri. "Zebrafish swimming in the flow: a particle image velocimetry study." PeerJ 5 (November 14, 2017): e4041. http://dx.doi.org/10.7717/peerj.4041.
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Zebrafish is emerging as a species of choice for the study of a number of biomechanics problems, including balance development, schooling, and neuromuscular transmission. The precise quantification of the flow physics around swimming zebrafish is critical toward a mechanistic understanding of the complex swimming style of this fresh-water species. Although previous studies have elucidated the vortical structures in the wake of zebrafish swimming in placid water, the flow physics of zebrafish swimming against a water current remains unexplored. In an effort to illuminate zebrafish swimming in a dynamic environment reminiscent of its natural habitat, we experimentally investigated the locomotion and hydrodynamics of a single zebrafish swimming in a miniature water tunnel using particle image velocimetry. Our results on zebrafish locomotion detail the role of flow speed on tail beat undulations, heading direction, and swimming speed. Our findings on zebrafish hydrodynamics offer a precise quantification of vortex shedding during zebrafish swimming and demonstrate that locomotory patterns play a central role on the flow physics. This knowledge may help clarify the evolutionary advantage of burst and cruise swimming movements in zebrafish.
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Hardianti, Mira, Ari Yuniarto, and Patonah Hasimun. "Review: Zebrafish (Danio Rerio) Sebagai Model Obesitas dan Diabetes Melitus Tipe 2." Jurnal Sains Farmasi & Klinis 8, no. 2 (August 6, 2021): 69. http://dx.doi.org/10.25077/jsfk.8.2.69-79.2021.
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Zebrafish (Danio rerio) yang sebelumnya disebut Brachydanio rerio merupakan spesies ikan air tawar tropis yang berukuran kecil yang berasal dari Asia Selatan dan Sungai Gangga di India. Zebrafish bisa diaplikasikan pada model obesitas dan diabetes melitus tipe 2 (DMT2). Obesitas merupakan pemicu terjadinya DMT2. Tujuan dari review artikel ini dapat memberikan informasi dan meningkatkan pemahaman pembaca bahwa zebrafish merupakan model yang ideal untuk obesitas dan DMT2. Metode penulisan review artikel ini dilakukan dengan penelusuran artikel ilmiah terpublikasi bertaraf nasional dan internasional dengan menggunakan kata kunci berupa “Obesity, zebrafish, model”, “Hyperglycemia, zebrafish, model” atau “zebrafish for hyperglycemia” dan “Diabetes melitus type 2, zebrafish, model”. Zebrafish model obesitas dan DMT2 yang diinduksi makanan dan obat menunjukkan keunggulan menjadi obesitas seperti pada manusia dan mamalia lain. Selain itu, zebrafish yang diinduksi dengan metode perendaman glukosa diperoleh hasil meningkatnya kadar glukosa darah dan gangguan respon terhadap insulin eksogen. Pemberian alternatif terapi dari bahan alam seperti GTE, palmaria mollis, diosgenin, bubuk kayu manis pada zebrafish memberikan hasil yang cukup baik sehingga bisa dipertimbangkan sebagai alternatif model hewan lain.
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Vorjohann, Silja, Frédérique Béna, Alexandre Fort, Richard J. Fish, and Marguerite Neerman-Arbez. "Developmental expression and organisation of fibrinogen genes in the zebrafish." Thrombosis and Haemostasis 107, no. 01 (2012): 158–66. http://dx.doi.org/10.1160/th11-04-0221.
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SummaryThe zebrafish is a model organism for studying vertebrate development and many human diseases. Orthologues of the majority of human coagulation factors are present in zebrafish, including fibrinogen. As a first step towards using zebrafish to model human fibrinogen disorders, we cloned the zebrafish fibrinogen cDNAs and made in situ hybridisations and quantitative reverse transcription-polymerase chain reactions (qRT-PCR) to detect zebrafish fibrinogen mRNAs. Prior to liver development or blood flow we detected zebrafish fibrinogen expression in the embryonic yolk syncytial layer and then in the early cells of the developing liver. While human fibrinogen is encoded by a three-gene, 50 kilobase (kb) cluster on chromosome 4 (FGB-FGA-FGG), recent genome assemblies showed that the zebrafish fgg gene appears distanced from fga and fgb, which we confirmed by in situ hybridisation. The zebrafish fibrinogen Bβ and γ protein chains are conserved at over 50% of amino acid positions, compared to the human polypeptides. The zebrafish Aα chain is less conserved and its C-terminal region is nearly 200 amino acids shorter than human Aα. We generated transgenic zebrafish which express a green fluorescent protein reporter gene under the control of a 1.6 kb regulatory region from zebrafish fgg. Transgenic embryos showed strong fluorescence in the developing liver, mimicking endogenous fibrinogen expression. This regulatory sequence can now be used for overexpression of transgenes in zebrafish hepatocytes. Our study is a proof-of-concept step towards using zebrafish to model human disease linked to fibrinogen gene mutations.
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Jin, Eun-Jung, and Giselle Thibaudeau. "Involvement of a LiCl-Induced Phosphoprotein in Pigmentation of the Embryonic Zebrafish (Danio rerio)." Journal of Life Science 18, no. 9 (September 30, 2008): 1219–24. http://dx.doi.org/10.5352/jls.2008.18.9.1219.
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Mao, Yousheng, Kwang-Heum Hong, Weifang Liao, Li Li, Seong-Jin Kim, Yinyi Xiong, In-Koo Nam, Seong-Kyu Choe, and Seong-Ae Kwak. "Generation of a Novel Transgenic Zebrafish for Studying Adipocyte Development and Metabolic Control." International Journal of Molecular Sciences 22, no. 8 (April 13, 2021): 3994. http://dx.doi.org/10.3390/ijms22083994.
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Zebrafish have become a popular animal model for studying various biological processes and human diseases. The metabolic pathways and players conserved among zebrafish and mammals facilitate the use of zebrafish to understand the pathological mechanisms underlying various metabolic disorders in humans. Adipocytes play an important role in metabolic homeostasis, and zebrafish adipocytes have been characterized. However, a versatile and reliable zebrafish model for long-term monitoring of adipose tissues has not been reported. In this study, we generated stable transgenic zebrafish expressing enhanced green fluorescent protein (EGFP) in adipocytes. The transgenic zebrafish harbored adipose tissues that could be detected using GFP fluorescence and the morphology of single adipocyte could be investigated in vivo. In addition, we demonstrated the applicability of this model to the long-term in vivo imaging of adipose tissue development and regulation based on nutrition. The transgenic zebrafish established in this study may serve as an excellent tool to advance the characterization of white adipose tissue in zebrafish, thereby aiding the development of therapeutic interventions to treat metabolic diseases in humans.
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Bourque, Caitlin, and Yariv Houvras. "Hooked on zebrafish: insights into development and cancer of endocrine tissues." Endocrine-Related Cancer 18, no. 5 (June 1, 2011): R149—R164. http://dx.doi.org/10.1530/erc-11-0099.
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Zebrafish is emerging as a unique model organism for studying cancer genetics and biology. For several decades zebrafish have been used to study vertebrate development, where they have made important contributions to understanding the specification and differentiation programs in many tissues. Recently, zebrafish studies have led to important insights into thyroid development, and have been used to model endocrine cancer. Zebrafish possess a unique set of attributes that make them amenable to forward and reverse genetic approaches. Zebrafish embryos develop rapidly and can be used to study specific cell lineages or the effects of chemicals on pathways or tissue development. In this review, we highlight the structure and function of endocrine organs in zebrafish and outline the major achievements in modeling cancer. Our goal is to familiarize readers with the zebrafish as a genetic model system and propose opportunities for endocrine cancer research in zebrafish.
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Oakes, James A., Lise Barnard, Karl-Heinz Storbeck, Vincent T. Cunliffe, and Nils P. Krone. "11β-Hydroxylase loss disrupts steroidogenesis and reproductive function in zebrafish." Journal of Endocrinology 247, no. 2 (November 2020): 197–212. http://dx.doi.org/10.1530/joe-20-0160.
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The roles of androgens in male reproductive development and function in zebrafish are poorly understood. To investigate this topic, we employed CRISPR/Cas9 to generate cyp11c1 (11β-hydroxylase) mutant zebrafish lines. Our study confirms recently published findings from a different cyp11c1−/− mutant zebrafish line, and also reports novel aspects of the phenotype caused by loss of Cyp11c1 function. We report that Cyp11c1-deficient zebrafish display predominantly female secondary sex characteristics, but may possess either ovaries or testes. Moreover, we observed that cyp11c1−/− mutant male zebrafish are profoundly androgen- and cortisol-deficient. These results provide further evidence that androgens are dispensable for testis formation in zebrafish, as has been demonstrated previously in androgen-deficient and androgen-resistant zebrafish. Herein, we show that the testes of cyp11c1−/− mutant zebrafish exhibit a disorganised tubular structure; and for the first time demonstrate that the spermatic ducts, which connect the testes to the urogenital orifice, are severely hypoplastic in androgen-deficient zebrafish. Furthermore, we show that spermatogenesis and characteristic breeding behaviours are impaired in cyp11c1−/− mutant zebrafish. Expression of nanos2, a type A spermatogonia marker, was significantly increased in the testes of Cyp11c1-deficient zebrafish, whereas expression of markers for later stages of spermatogenesis was significantly decreased. These observations indicate that in zebrafish, production of type A spermatogonia is androgen-independent, but differentiation of type A spermatogonia is an androgen-dependent process. Overall, our results demonstrate that whilst androgens are not required for testis formation, they play important roles in determining secondary sexual characteristics, proper organisation of seminiferous tubules, and differentiation of male germ cells.
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McGonnell, I. M., and R. C. Fowkes. "Fishing for gene function – endocrine modelling in the zebrafish." Journal of Endocrinology 189, no. 3 (June 2006): 425–39. http://dx.doi.org/10.1677/joe.1.06683.
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The use of zebrafish (Danio rerio) in scientific research is growing rapidly. It initially became popular as a model of vertebrate development because zebrafish embryos develop rapidly and are transparent. In the past 5 years, the sequencing of the zebrafish genome has increased the profile of zebrafish research even further, expanding into other areas such as pharmacology, cancer research and drug discovery. The use of zebrafish in endocrine research has mainly been confined to the study of the development of endocrine organs. However, it is likely to be a useful model in other areas of endocrinology, as there are a wide variety of both forward and reverse genetic techniques that can be employed in the zebrafish to decipher gene function in disease states. In this review, we compare the endocrine system of the zebrafish to mouse and human, demonstrating that the systems are sufficiently similar for zebrafish to be employed as a model for endocrine research. We subsequently review the repertoire of genetic techniques commonly employed in the zebrafish model to understand gene function in vertebrate development and disease. We anticipate that the use of these techniques will make the zebrafish a prominent model in endocrine research in the coming years.
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Wiggenhauser, Lucas Moritz, and Jens Kroll. "Vascular Damage in Obesity and Diabetes: Highlighting Links Between Endothelial Dysfunction and Metabolic Disease in Zebrafish and Man." Current Vascular Pharmacology 17, no. 5 (August 1, 2019): 476–90. http://dx.doi.org/10.2174/1570161116666181031101413.
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Endothelial dysfunction is an initial pathophysiological mechanism of vascular damage and is further recognized as an independent predictor of negative prognosis in diabetes-induced micro- and macrovascular complications. Insight into the capability of zebrafish to model metabolic disease like obesity and type II diabetes has increased and new evidence on the induction of vascular pathologies in zebrafish through metabolic disease is available. Here, we raise the question, if zebrafish can be utilized to study the initial impairments of vascular complications in metabolic disorders. In this review, we focus on the advances made to develop models of obesity and type II diabetes in zebrafish, discuss the key points and characteristics of these models, while highlighting the available information linked to the development of endothelial dysfunction in zebrafish and man. We show that larval and adult zebrafish develop metabolic dysregulation in the settings of obesity and diabetes, exhibiting pathophysiological mechanisms, which mimic the human condition. The most important genes related to endothelial dysfunction are present in zebrafish and further display similar functions as in mammals. Several suggested contributors to endothelial dysfunction found in these models, namely hyperinsulinaemia, hyperglycaemia, hyperlipidaemia and hyperleptinaemia are highlighted and the available data from zebrafish are summarised. Many underlying processes of endothelial dysfunction in obesity and diabetes are fundamentally present in zebrafish and provide ground for the assumption, that zebrafish can develop endothelial dysfunction. Conservation of basic biological mechanisms is established for zebrafish, but focused investigation on the subject is now needed as validation and particularly more research is necessary to understand the differences between zebrafish and man. The available data demonstrate the relevance of zebrafish as a model for metabolic disease and their ability to become a proponent for the investigation of vascular damage in the settings of obesity and diabetes.
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Cheng, Ruey-Kuang, Suresh J. Jesuthasan, and Trevor B. Penney. "Zebrafish forebrain and temporal conditioning." Philosophical Transactions of the Royal Society B: Biological Sciences 369, no. 1637 (March 5, 2014): 20120462. http://dx.doi.org/10.1098/rstb.2012.0462.
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The rise of zebrafish as a neuroscience research model organism, in conjunction with recent progress in single-cell resolution whole-brain imaging of larval zebrafish, opens a new window of opportunity for research on interval timing. In this article, we review zebrafish neuroanatomy and neuromodulatory systems, with particular focus on identifying homologies between the zebrafish forebrain and the mammalian forebrain. The neuroanatomical and neurochemical basis of interval timing is summarized with emphasis on the potential of using zebrafish to reveal the neural circuits for interval timing. The behavioural repertoire of larval zebrafish is reviewed and we demonstrate that larval zebrafish are capable of expecting a stimulus at a precise time point with minimal training. In conclusion, we propose that interval timing research using zebrafish and whole-brain calcium imaging at single-cell resolution will contribute to our understanding of how timing and time perception originate in the vertebrate brain from the level of single cells to circuits.
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Liu, Yunxing, Fang Liang, Zijiong Dong, Song Li, Jianmin Ye, and Wei Qin. "Genome Editing in Zebrafish by ScCas9 Recognizing NNG PAM." Cells 10, no. 8 (August 16, 2021): 2099. http://dx.doi.org/10.3390/cells10082099.
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The CRISPR/Cas9 system has been widely used for gene editing in zebrafish. However, the required NGG protospacer adjacent motif (PAM) of Streptococcus pyogenes Cas9 (SpCas9) notably restricts the editable range of the zebrafish genome. Recently, Cas9 from S. canis (ScCas9), which has a more relaxed 5′-NNG-3′ PAM, was reported to have activities in human cells and plants. However, the editing ability of ScCas9 has not been tested in zebrafish. Here we characterized and optimized the activity of ScCas9 in zebrafish. Delivered as a ribonucleoprotein complex, ScCas9 can induce mutations in zebrafish. Using the synthetic modified crRNA:tracrRNA duplex instead of in vitro-transcribed single guide RNA, the low activity at some loci were dramatically improved in zebrafish. As far as we know, our work is the first report on the evaluation of ScCas9 in animals. Our work optimized ScCas9 as a new nuclease for targeting relaxed NNG PAMs for zebrafish genome editing, which will further improve genome editing in zebrafish.
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Gart, Michael S., Ramy A. Shoela, Joanna P. Tomaszewski, Tatiana Favelevic, Jolanta M. Topczewska, and Arun K. Gosain. "Zebrafish." Plastic and Reconstructive Surgery 134 (October 2014): 2–3. http://dx.doi.org/10.1097/01.prs.0000455315.23008.3f.
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Raby, Ludivine, Pamela Völkel, Xuefen Le Bourhis, and Pierre-Olivier Angrand. "Genetic Engineering of Zebrafish in Cancer Research." Cancers 12, no. 8 (August 4, 2020): 2168. http://dx.doi.org/10.3390/cancers12082168.
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Zebrafish (Danio rerio) is an excellent model to study a wide diversity of human cancers. In this review, we provide an overview of the genetic and reverse genetic toolbox allowing the generation of zebrafish lines that develop tumors. The large spectrum of genetic tools enables the engineering of zebrafish lines harboring precise genetic alterations found in human patients, the generation of zebrafish carrying somatic or germline inheritable mutations or zebrafish showing conditional expression of the oncogenic mutations. Comparative transcriptomics demonstrate that many of the zebrafish tumors share molecular signatures similar to those found in human cancers. Thus, zebrafish cancer models provide a unique in vivo platform to investigate cancer initiation and progression at the molecular and cellular levels, to identify novel genes involved in tumorigenesis as well as to contemplate new therapeutic strategies.
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Huang, Shih-Hao, Chia-Wei Tsao, and Yan-Hung Fang. "A Miniature Intermittent-Flow Respirometry System with a 3D-Printed, Palm-Sized Zebrafish Treadmill for Measuring Rest and Activity Metabolic Rates." Sensors 20, no. 18 (September 7, 2020): 5088. http://dx.doi.org/10.3390/s20185088.
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Zebrafish are a preferred vertebrate model for evaluating metabolism during development, and for toxicity studies. However, commercially available intermittent-flow respirometry systems (IFRS) do not provide a suitable zebrafish-scaled swimming tunnel with a low water volume and proper flow velocities. We developed a miniature IFRS (mIFRS) with a 3D-printed, palm-sized zebrafish treadmill for measuring the swimming ability and metabolic rate of a single one- or three-month-old zebrafish with and without toxicity treatment. The 3D-printed zebrafish treadmill consists of discrete components assembled together which enables the provision of a temporary closed circulating water flow. The results showed that three-month-old zebrafish of normal physiological status had higher energetic efficiency and could swim at a higher critical swimming speed (Ucrit) of 16.79 cm/s with a lower cost of transport (COTopt) of 0.11 μmol g−1m−1. However, for a single three-month-old zebrafish treated with an antibacterial agent, Ucrit decreased to 45% of normal zebrafish and the COTopt increased to 0.24 μmol g−1m−1, due to the impairment of mitochondria. Our mIFRS provides a low-cost, portable, and readily adaptable tool for studying the swimming performance and energetic metabolism of zebrafish.
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Zhang, Mingming, Wei Sun, Jianan Du, Yawei Gou, Lingling Liu, Ruonan Wang, and Xuesong Xu. "Protective Effect of Metformin on Sepsis Myocarditis in Zebrafish." Dose-Response 18, no. 3 (July 1, 2020): 155932582093854. http://dx.doi.org/10.1177/1559325820938543.
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Purpose: We found in previous study that metformin could treat sepsis myocarditis in a mouse model. We employed the zebrafish model organism to investigate the effect of metformin on sepsis myocarditis. Methods and Results: Wild-type zebrafish was used to establish a sepsis myocarditis model and combined with image software analysis and cytokine detection, the protective dose of metformin was determined. The results showed that immersion with Escherichia coli could cause 75% mortality in zebrafish and make larvae appear as characteristics of severe sepsis myocarditis. Pretreatment with 10 mM metformin for 3 hours could effectively reduce heart congestion and swelling in zebrafish with sepsis myocarditis and increased the heart rate. It could reduce the mortality and prolong the survival time of zebrafish with sepsis myocarditis; Tg(mpx: EGFP) transgenic zebrafish were adopted to explore the number of neutrophils in zebrafish heart before and after metformin protection, and metformin could maintain the number of neutrophils in zebrafish heart; quantitative real-time reverse transcription–polymerase chain reaction showed that metformin could reduce the expression of pro-inflammatory factors, tumor necrosis factor-α and interleukin (IL)-6, and could promote the anti-inflammatory factor, transforming growth factor-β and IL-10 expression. Conclusion: We established a zebrafish sepsis myocarditis model and applied metformin in advance to provide a protective effect on the zebrafish heart.
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Chevrette, Mario, Lucille Joly, Patricia Tellis, and Marc Ekker. "Contribution of zebrafish-mouse cell hybrids to the mapping of the zebrafish genome." Biochemistry and Cell Biology 75, no. 5 (October 1, 1997): 641–49. http://dx.doi.org/10.1139/o97-082.
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The zebrafish, Danio rerio, is becoming an increasingly popular model for the study of vertebrate development. Indeed, the biology of the fish offers great advantages for such studies. The life cycle of the zebrafish is relatively short (2-3 months) and the embryos develop outside the mother, facilitating the visualization of any mutated phenotype. At present, more than 1000 embryonic mutations have been reported. However, until recently, there was no physical or genetic map for this organism. In an effort to generate such a map, we have produced and characterized a panel of zebrafish-mouse cell hybrids. We have used whole-cell fusion to transfer zebrafish chromosomes from two different zebrafish cell lines into mouse recipient cells, thus generating more than 100 hybrids. Using fluorescence in situ hybridization and polymerase chain reaction analysis, we have determined the zebrafish chromosome composition of these hybrids. Here we report that elements from the 25 linkage groups of the zebrafish genome are present in our hybrids. These hybrids could identify the chromosomal location of genes affected in zebrafish mutants.
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Spitsbergen, Jan M., and Michael L. Kent. "The State of the Art of the Zebrafish Model for Toxicology and Toxicologic Pathology Research—Advantages and Current Limitations." Toxicologic Pathology 31, no. 1_suppl (January 2003): 62–87. http://dx.doi.org/10.1080/01926230390174959.
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The zebrafish (Danio rerio ) is now the pre-eminent vertebrate model system for clarification of the roles of specific genes and signaling pathways in development. The zebrafish genome will be completely sequenced within the next 1—2 years. Together with the substantial historical database regarding basic developmental biology, toxicology, and gene transfer, the rich foundation of molecular genetic and genomic data makes zebrafish a powerful model system for clarifying mechanisms in toxicity. In contrast to the highly advanced knowledge base on molecular developmental genetics in zebrafish, our database regarding infectious and noninfectious diseases and pathologic lesions in zebrafish lags far behind the information available on most other domestic mammalian and avian species, particularly rodents. Currently, minimal data are available regarding spontaneous neoplasm rates or spontaneous aging lesions in any of the commonly used wild-type or mutant lines of zebrafish. Therefore, to fully utilize the potential of zebrafish as an animal model for understanding human development, disease, and toxicology we must greatly advance our knowledge on zebrafish diseases and pathology.
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Markovich, Daniel, Alessandro Romano, Carlo Storelli, and Tiziano Verri. "Functional and structural characterization of the zebrafish Na+-sulfate cotransporter 1 (NaS1) cDNA and gene (slc13a1)." Physiological Genomics 34, no. 3 (August 2008): 256–64. http://dx.doi.org/10.1152/physiolgenomics.90234.2008.
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Sulfate plays an essential role during growth, development and cellular metabolism. In this study, we characterized the function and structure of the zebrafish ( Danio rerio) Na+-sulfate cotransporter 1 (NaS1) cDNA and gene ( slc13a1). Zebrafish NaS1 encodes a protein of 583 amino acids with 13 putative transmembrane domains. Expression of zebrafish NaS1 protein in Xenopus oocytes led to Na+-sulfate cotransport, which was significantly inhibited by thiosulfate, selenate, molybdate, and tungstate. Zebrafish NaS1 transport kinetics were: Vmax = 1,731.670 ± 92.853 pmol sulfate/oocyte·hour and Km = 1.414 ± 0.275 mM for sulfate and Vmax = 307.016 ± 32.992 pmol sulfate/oocyte·hour, Km = 24.582 ± 4.547 mM and n (Hill coefficient) = 1.624 ± 0.354 for sodium. Zebrafish NaS1 mRNA is developmentally expressed in embryos from day 1 postfertilization and in the intestine, kidney, brain, and eye of adult zebrafish. The zebrafish NaS1 gene slc13a1 contains 15 exons spanning 8,716 bp. Characterization of the zebrafish NaS1 contributes to a greater understanding of sulfate transporters in a well-defined genetic model and will allow the elucidation of evolutionary and functional relationships among vertebrate sulfate transporters.
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Morikane, Daizo, Liqing Zang, and Norihiro Nishimura. "Evaluation of the Percutaneous Absorption of Drug Molecules in Zebrafish." Molecules 25, no. 17 (August 31, 2020): 3974. http://dx.doi.org/10.3390/molecules25173974.
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In recent decades, zebrafish (Danio rerio) has become a widely used vertebrate animal model for studying development and human diseases. However, studies on skin medication using zebrafish are rare. Here, we developed a novel protocol for percutaneous absorption of molecules via the zebrafish tail skin, by applying a liquid solution directly, or using a filter paper imbibed with a chemical solution (coating). Human skin is capable of absorbing felbinac and loxoprofen sodium hydrate (LSH), but not glycyrrhetinic acid (GA) and terbinafine hydrochloride (TH). To evaluate the possibility and the quality of transdermal absorption in zebrafish, we transdermally administered these four drugs to zebrafish. Pharmacokinetics showed that felbinac was present in the blood of zebrafish subjected to all administration methods. Felbinac blood concentrations peaked at 2 h and disappeared 7 h after administration. GA was not detected following transdermal administrations, but was following exposure. LSH was not found in the circulatory system after transdermal administration, but TH was. A dose-response correlation was observed for felbinac blood concentration. These findings suggest that zebrafish are capable of absorbing drug molecules through their skin. However, the present data cannot demonstrate that zebrafish is a practical model to predict human skin absorption. Further systemic studies are needed to observe the correlations in percutaneous absorption between humans and zebrafish.
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Lin, Jen-Chieh, Shing Hu, Pei-Hung Ho, Hwei-Jan Hsu, John H. Postlethwait, and Bon-chu Chung. "Two Zebrafish hsd3b Genes Are Distinct in Function, Expression, and Evolution." Endocrinology 156, no. 8 (August 1, 2015): 2854–62. http://dx.doi.org/10.1210/en.2014-1584.
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Abstract HSD3B catalyzes the synthesis of δ4 steroids such as progesterone in the adrenals and gonads. Individuals lacking HSD3B2 activity experience congenital adrenal hyperplasia with imbalanced steroid synthesis. To develop a zebrafish model of HSD3B deficiency, we characterized 2 zebrafish hsd3b genes. Our phylogenetic and conserved synteny analyses showed that the tandemly duplicated human HSD3B1 and HSD3B2 genes are coorthologs of zebrafish hsd3b1 on chromosome 9 (Dre9), whereas the gene called hsd3b2 resides on Dre20 in an ancestral chromosome segment, from which its ortholog was lost in the tetrapod lineage. Zebrafish hsd3b1(Dre 9) was expressed in adult gonads and headkidney, which contains interrenal glands, the zebrafish counterpart of the tetrapod adrenal. Knockdown of hsd3b1(Dre 9) caused the interrenal and anterior pituitary to expand and pigmentation to increase, resembling human HSD3B2 deficiency. The zebrafish hsd3b2(Dre 20) gene was expressed in zebrafish early embryos as maternal transcripts that disappeared 1 day after fertilization. Morpholino inactivation of hsd3b2(Dre 20) led to embryo elongation, which was rescued by the injection of hsd3b2 mRNA. Thus, zebrafish hsd3b2(Dre 20) evolved independently of hsd3b1(Dre 9) with a morphogenetic function during early embryogenesis. Zebrafish hsd3b1(Dre 9), on the contrary, functions like mammalian HSD3B2, whose deficiency leads to congenital adrenal hyperplasia.
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Baek, Ji Sun, Longhou Fang, Andrew C. Li, and Yury I. Miller. "Ezetimibe and Simvastatin Reduce Cholesterol Levels in Zebrafish Larvae Fed a High-Cholesterol Diet." Cholesterol 2012 (May 30, 2012): 1–5. http://dx.doi.org/10.1155/2012/564705.
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Cholesterol-fed zebrafish is an emerging animal model to study metabolic, oxidative, and inflammatory vascular processes relevant to pathogenesis of human atherosclerosis. Zebrafish fed a high-cholesterol diet (HCD) develop hypercholesterolemia and are characterized by profound lipoprotein oxidation and vascular lipid accumulation. Using optically translucent zebrafish larvae has the advantage of monitoring vascular pathology and assessing the efficacy of drug candidates in live animals. Thus, we investigated whether simvastatin and ezetimibe, the principal drugs used in management of hypercholesterolemia in humans, would also reduce cholesterol levels in HCD-fed zebrafish larvae. We found that ezetimibe was well tolerated by zebrafish and effectively reduced cholesterol levels in HCD-fed larvae. In contrast, simvastatin added to water was poorly tolerated by zebrafish larvae and, when added to food, had little effect on cholesterol levels in HCD-fed larvae. Combination of low doses of ezetimibe and simvastatin had an additive effect in reducing cholesterol levels in zebrafish. These results suggest that ezetimibe exerts in zebrafish a therapeutic effect similar to that in humans and that the hypercholesterolemic zebrafish can be used as a low-cost and informative model for testing new drug candidates and for investigating mechanisms of action for existing drugs targeting dyslipidemia.
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Biga, P. R., and F. W. Goetz. "Zebrafish and giant danio as models for muscle growth: determinate vs. indeterminate growth as determined by morphometric analysis." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 291, no. 5 (November 2006): R1327—R1337. http://dx.doi.org/10.1152/ajpregu.00905.2005.
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The zebrafish has become an important genetic model, but their small size makes them impractical for traditional physiological studies. In contrast, the closely related giant danio is larger and can be utilized for physiological studies that can also make use of the extensive zebrafish genomic resources. In addition, the giant danio and zebrafish appear to exhibit different growth types, indicating the potential for developing a comparative muscle growth model system. Therefore, the present study was conducted to compare and characterize the muscle growth pattern of zebrafish and giant danio. Morphometric analyses demonstrated that giant danio exhibit an increased growth rate compared with zebrafish, starting as early as 2 wk posthatch. Total myotome area, mean fiber area, and total fiber number all exhibited positive correlations with larvae length in giant danio but not in zebrafish. Morphometric analysis of giant danio and zebrafish larvae demonstrated faster, more efficient growth in giant danio larvae. Similar to larger teleosts, adult giant danio exhibited increased growth rates in response to growth hormone, suggesting that giant danio exhibit indeterminate growth. In contrast, adult zebrafish do not exhibit mosaic hyperplasia, nor do they respond to growth hormone, suggesting they exhibit determinate growth like mammals. These results demonstrate that giant danio and zebrafish can be utilized as a direct comparative model system for muscle growth studies, with zebrafish serving as a model organism for determinate growth and giant danio for indeterminate growth.
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Delomas, Thomas A., and Konrad Dabrowski. "Zebrafish embryonic development is induced by carp sperm." Biology Letters 12, no. 11 (November 2016): 20160628. http://dx.doi.org/10.1098/rsbl.2016.0628.
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Haploid gynogenetic screens increase the efficiency of forward genetic screens and linkage analysis in fish. Typically, UV-irradiated zebrafish sperm is used to activate zebrafish oocytes for haploid screens. We describe the use of UV-irradiated common carp sperm to activate haploid gynogenetic zebrafish development. Carp × zebrafish hybrids are shown to have a characteristic set of features during embryonic development and exhibit functional development of several tissues (muscle, heart and nervous system). Hybrids become inviable past the embryonic stages. This technique eliminates the possibility of incompletely irradiated zebrafish spermatozoa contaminating haploid progenies. While developing this protocol, one unique zebrafish female was identified which, upon insemination with UV-irradiated carp spermatozoa, repeatedly displayed spontaneous diploidization of the maternal chromosomes in her offspring.
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Chao, Chun-Cheih, Po-Chen Hsu, Chung-Feng Jen, I.-Hui Chen, Chieh-Huei Wang, Hau-Chien Chan, Pei-Wen Tsai, et al. "Zebrafish as a Model Host for Candida albicans Infection." Infection and Immunity 78, no. 6 (March 22, 2010): 2512–21. http://dx.doi.org/10.1128/iai.01293-09.
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ABSTRACTIn this work, the zebrafish model organism was developed to obtain a minivertebrate host system for aCandida albicansinfection study. We demonstrated thatC. albicanscan colonize and invade zebrafish at multiple anatomical sites and kill the fish in a dose-dependent manner. Inside zebrafish, we monitored the progression of theC. albicansyeast-to-hypha transition by tracking morphogenesis, and we monitored the corresponding gene expression of the pathogen and the early host immune response. We performed a zebrafish survival assay with differentC. albicansstrains (SC5314, ATCC 10231, anhgc1mutant, and acph1/efg1double mutant) to determine each strain's virulence, and the results were similar to findings reported in previous mouse model studies. Finally, using zebrafish embryos, we monitoredC. albicansinfection and visualized the interaction between pathogen and host myelomonocytic cellsin vivo. Taken together, the results of this work demonstrate that zebrafish can be a useful host model to studyC. albicanspathogenesis, and they highlight the advantages of using the zebrafish model in future invasive fungal research.
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Kumar, Madan Mohan Udaya, Boon Chuan Low, and Zhiyuan Gong. "Signal Transduction in Hepatocellular Carcinoma: Insights from Zebrafish." Proceedings of the Singapore National Academy of Science 14, no. 01 (September 2020): 47–58. http://dx.doi.org/10.1142/s2591722620400049.
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The zebrafish has been an increasingly popular animal model for human diseases as it offers the combined advantages compared to various animal models and cell based assays; in particular, the feasibility of high throughput studies as an economical vertebrate model. In this past decade, we and several other laboratories have developed various hepatocellular carcinoma (HCC) models using the zebrafish and demonstrated the conservation of HCC between zebrafish and human at both histopathological and molecular levels. In this review, we focus on the conservation of signal transductions during hepatocarcinogenesis between zebrafish and human. Based on existing zebrafish HCC models, indeed many alterations of signal pathways that cause human liver cancers can also result in HCC in zebrafish, such as Ras pathway, EGFR pathway, Wng/β-catenin pathway, TGF-β pathway, PI3K/AKT pathway, JAK/STAT pathway, Hippo pathway, src tyrosine kinase pathway, etc. In future, zebrafish may be used for better quantification of signaling molecules and thus to aid development of more effective therapeutic methods.
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Belyaeva, N. F., V. N. Kashirtseva, N. V. Medvedeva, Yu Yu Khudoklinova, O. M. Ipatova, and A. I. Archakov. "Zebrafish as a model system for biomedical studies." Biomeditsinskaya Khimiya 56, no. 1 (January 2010): 120–31. http://dx.doi.org/10.18097/pbmc20105601120.
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Zebrafish (Danio rerio) are now firmly established as a powerful research model for many areas of biology and medicine. Here, we review some achievements of zebrafish - based assays for modeling human diseases and for drug discovery and development. For drug discovery, zebrafish are especially valuable in the earlier stages of research as they provide a model organism to demonstrate a new treatment's efficacy and toxicity before more costly mammalian models are used. This review provides examples of compounds known to be toxic to humans that have been demonstrated to functional similarly in zebrafish. Major advantages of zebrafish embryons are that they are readily permeable to small molecules added to their incubation medium and the transparent chorion enables the easy observation of development. Assay of acute toxicity (LC50 estimation) in embryos can also include the screening for developmental disorders as an indicator of teratogenic effects. We used zebrafish for toxicity testing of new drugs on the base of phospholipid nanoparticles. The organization of the genome and the pathways controlling signal transduction appear to be highly conserved between zebrafish and humans that allow using zebrafish for modeling of human diseases some examples of which are illustrated in this paper.
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Yin, Heng, Jianwei Wang, Mao Wu, Yong Ma, Shanfu Wang, and Qiuju Su. "Preventive Effects of Evodiamine on Dexamethasone-Induced Osteoporosis in Zebrafish." BioMed Research International 2019 (January 22, 2019): 1–6. http://dx.doi.org/10.1155/2019/5859641.
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The aim of this study was to investigate the effect of evodiamine (EV) on dexamethasone-induced osteoporosis in zebrafish. Zebrafish larvae were exposed to different concentrations of dexamethasone to obtain the osteoporosis in zebrafish. Calcium, phosphorus, and alizarin red staining determination were performed to evaluate the effects of EV on bone mineralization. Alkaline phosphatase (ALP), hydroxyproline (HP), and tartrate resistant acid phosphatase (TRAP) were also measured by commercial kits. The expression of MMP3-OPN-MAPK pathway in zebrafish was measured by Western blot. RT-PCR was used to determine mRNA levels of MMP3, OPN, and MAPK. EV could significantly increase the content of calcium and phosphorus. The results of alizarin red staining showed that EV could significantly increase the calcium sink of horse fish, increasing the area of bone formation. EV could increase the content of hydroxyproline in zebrafish. EV also increased ALP and TRAP in zebrafish. Western blot and RT-PCR results showed that EV restored the MMP3-OPN-MAPK pathway in zebrafish. In conclusion, we found that EV can alleviate dexamethasone-induced osteoporosis in zebrafish. The mechanism is related to activating MMP3-OPN-MAPK pathway and then activating bone remodeling.
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Chng, Hui Ting, Han Kiat Ho, Chun Wei Yap, Siew Hong Lam, and Eric Chun Yong Chan. "An Investigation of the Bioactivation Potential and Metabolism Profile of Zebrafish versus Human." Journal of Biomolecular Screening 17, no. 7 (May 29, 2012): 974–86. http://dx.doi.org/10.1177/1087057112447305.
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The zebrafish model has been increasingly explored as an alternative model for toxicity screening of pharmaceutical drugs. However, little is understood about the bioactivation of drug to reactive metabolite and phase I and II metabolism of chemical in zebrafish as compared with human. The primary aim of our study was to establish the bioactivation potential of zebrafish using acetaminophen as a probe substrate. Our secondary aim was to perform metabolite profiling experiments on testosterone, a CYP3A probe substrate, in zebrafish and compare the metabolite profiles with that of human. The glutathione trapping assay of N-acetyl- p-benzoquinone imine demonstrated that zebrafish generates the same reactive metabolite as humans from the bioactivation of acetaminophen. Zebrafish possesses functional CYP3A4/5-like and UDP-glucuronosyltransferase metabolic activities on testosterone. Differential testosterone metabolism was observed among the two species. In silico docking studies suggested that the zebrafish CYP3A65 was responsible for the bioactivation of acetaminophen and phase I hydroxylation of testosterone. Our findings reinforce the need to further characterize the drug metabolism phenotype of zebrafish before the model can fully achieve its potential as an alternative toxicity screening model in drug research.
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Ye, Yongbin, Xiaojun Xu, and Qifa Liu. "Establishment of a Xenotransplantation Acute Myeloid Leukemia Model Using By Zebrafish." Blood 128, no. 22 (December 2, 2016): 5770. http://dx.doi.org/10.1182/blood.v128.22.5770.5770.
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Abstract Objecitve: To investigate the feasibility ofxenotransplant leukemia model using by zebrafish, and lay the foundation for the study of acute myeloid leukemia and the screening of the drugs for targeting therapy of acute myeloid leukemia. Methods: Acute myeloid leukemia cell line KG1a was labeled with red fluorescent dye-MitoRed, then, the labeled cells was injected into the yolk sac of zebrafish embryos. Morphology observation, cell counting and histopathological detection were used to analysis the infiltration and metastasis of KG-1a cells in zebrafish. Results: KG1a cells could proliferate and gradually spread to the entire abdominal cavity of the zebrafish After KG1a cells were injected into the yolk sac during 1-7 dpi, the results of cell counting in vitro also proved a signification proliferation of KG1a cells in zebrafish, suggesting that the implanted leukemia stem cells could survive, proliferate and spread in zebrafish. Further study detected the implanted cells could transfered to the liver of zebrafish, these cells showed the signature of KG1a cells by hematoxy -lin -eosin(HE) staining. Conclusions: Human acute myeloid leukemia cells KG1a could survive, proliferate and migrate in zebrafish, suggesting xenotransplant leukemia model using by zebrafish is feasible. This model may provide benefit for the study of acute myeloid leukemia and the screening of the drugs for targeting therapy of acute myeloid leukemia. Disclosures No relevant conflicts of interest to declare.
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Siregar, Petrus, Stevhen Juniardi, Gilbert Audira, Yu-Heng Lai, Jong-Chin Huang, Kelvin H. C. Chen, Jung-Ren Chen, and Chung-Der Hsiao. "Method Standardization for Conducting Innate Color Preference Studies in Different Zebrafish Strains." Biomedicines 8, no. 8 (August 3, 2020): 271. http://dx.doi.org/10.3390/biomedicines8080271.
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The zebrafish has a tetrachromatic vision that is able to distinguish ultraviolet (UV) and visible wavelengths. Recently, zebrafish color preferences have gained much attention because of the easy setup of the instrument and its usefulness to screen behavior-linked stimuli. However, several published papers dealing with zebrafish color preferences have contradicting results that underscore the importance of method standardization in this field. Different laboratories may report different results because of variations in light source, color intensity, and other parameters such as age, gender, container size, and strain of fish. In this study, we aim to standardize the color preference test in zebrafish by measuring light source position, light intensity, gender, age, animal size to space ratio, and animal strain. Our results showed that color preferences for zebrafish are affected by light position, age, strain, and social interaction of the fish, but not affected by fish gender. We validated that ethanol can significantly induce color preference alteration in zebrafish which may be related to anxiety and depression. We also explored the potential use of the optimized method to examine color preference ranking and index differences in various zebrafish strains and species, such as the tiger barb and glass catfish. In conclusion, zebrafish color preference screening is a powerful tool for high-throughput neuropharmacological applications and the standardized protocol established in this study provides a useful reference for the zebrafish research community.
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Lu, Xing, Yong Long, Xixi Li, Lang Zhang, Qing Li, Hua Wen, Shan Zhong, and Zongbin Cui. "Generation of Knockout and Transgenic Zebrafish to Characterize Abcc4 Functions in Detoxification and Efflux of Lead." International Journal of Molecular Sciences 22, no. 4 (February 19, 2021): 2054. http://dx.doi.org/10.3390/ijms22042054.
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Lead (Pb) is one of the major heavy metals that are toxic to vertebrates and usually considered as environmental pollutants. ABCC4/MRP4 is an organic anion transporter that mediates cellular efflux of a wide range of exogenous and endogenous compounds such as cyclic nucleotides and anti-cancer drugs; however, it remains unclear whether ABCC4 and its orthologs function in the detoxification and excretion of toxic lead. In this study, we found that the transcriptional and translational expression of zebrafish abcc4 was significantly induced under lead exposure in developing zebrafish embryos and adult tissues. Overexpression of zebrafish Abcc4 markedly decreased the cytotoxicity and accumulation of lead in pig renal proximal tubule cell line (LLC-PK1 cells). To further understand the functions of zebrafish Abcc4 in lead detoxification, the clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system was used to create an abcc4−/− mutant zebrafish line. In comparison with the wild-type (WT) zebrafish, the abcc4−/− mutants showed a higher death rate and lead accumulation upon exposure to lead. Furthermore, a stable abcc4-transgenic zebrafish line was successfully generated, which exerted stronger ability to detoxify and excrete lead than WT zebrafish. These findings indicate that zebrafish Abcc4 plays a crucial role in lead detoxification and cellular efflux and could be used as a potential biomarker to monitor lead contamination in a water environment.
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Phelan, Peter E., Meagan E. Pressley, P. Eckhard Witten, Mark T. Mellon, Sharon Blake, and Carol H. Kim. "Characterization of Snakehead Rhabdovirus Infection in Zebrafish (Danio rerio)." Journal of Virology 79, no. 3 (February 1, 2005): 1842–52. http://dx.doi.org/10.1128/jvi.79.3.1842-1852.2005.
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ABSTRACT The zebrafish, Danio rerio, has become recognized as a valuable model for the study of development, genetics, and toxicology. Recently, the zebrafish has been recognized as a useful model for infectious disease and immunity. In this study, the pathogenesis and antiviral immune response of zebrafish to experimental snakehead rhabdovirus (SHRV) infection was characterized. Zebrafish 24 h postfertilization to 30 days postfertilization were susceptible to infection by immersion in 106 50% tissue culture infective doses (TCID50) of SHRV/ml, and adult zebrafish were susceptible to infection by intraperitoneal (i.p.) injection of 105 TCID50 of SHRV/ml. Mortalities exceeded 40% in infected fish, and clinical presentation of infection included petechial hemorrhaging, redness of the abdomen, and erratic swim behavior. Virus reisolation and reverse transcription-PCR analysis of the viral nucleocapsid gene confirmed the presence of SHRV. Histological sections of moribund embryonic and juvenile fish revealed necrosis of the pharyngeal epithelium and liver, in addition to congestion of the swim bladder by cell debris. Histopathology in adult fish injected i.p. was confined to the site of injection. The antiviral response in zebrafish was monitored by quantitative real-time PCR analysis of zebrafish interferon (IFN) and Mx expression. IFN and Mx levels were elevated in zebrafish exposed to SHRV, although expression and intensity differed with age and route of infection. This study is the first to examine the pathogenesis of SHRV infection in zebrafish. Furthermore, this study is the first to describe experimental infection of zebrafish embryos with a viral pathogen, which will be important for future experiments involving targeted gene disruption and forward genetic screens.
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Zhao, Zhenkai, Gen Li, Qing Xiao, Hui-Rong Jiang, Gabriel Mbuta Tchivelekete, Xinhua Shu, and Hao Liu. "Quantification of the influence of drugs on zebrafish larvae swimming kinematics and energetics." PeerJ 8 (January 8, 2020): e8374. http://dx.doi.org/10.7717/peerj.8374.
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The use of zebrafish larvae has aroused wide interest in the medical field for its potential role in the development of new therapies. The larvae grow extremely quickly and the embryos are nearly transparent which allows easy examination of its internal structures using fluorescent imaging techniques. Medical treatment of zebrafish larvae can directly influence its swimming behaviours. These behaviour changes are related to functional changes of central nervous system and transformations of the zebrafish body such as muscle mechanical power and force variation, which cannot be measured directly by pure experiment observation. To quantify the influence of drugs on zebrafish larvae swimming behaviours and energetics, we have developed a novel methodology to exploit intravital changes based on observed zebrafish locomotion. Specifically, by using an in-house MATLAB code to process the recorded live zebrafish swimming video, the kinematic locomotion equation of a 3D zebrafish larvae was obtained, and a customised Computational Fluid Dynamics tool was used to solve the fluid flow around the fish model which was geometrically the same as experimentally tested zebrafish. The developed methodology was firstly verified against experiment, and further applied to quantify the fish internal body force, torque and power consumption associated with a group of normal zebrafish larvae vs. those immersed in acetic acid and two neuroactive drugs. As indicated by our results, zebrafish larvae immersed in 0.01% acetic acid display approximately 30% higher hydrodynamic power and 10% higher cost of transport than control group. In addition, 500 μM diphenylhydantoin significantly decreases the locomotion activity for approximately 50% lower hydrodynamic power, whereas 100 mg/L yohimbine has not caused any significant influences on 5 dpf zebrafish larvae locomotion. The approach has potential to evaluate the influence of drugs on the aquatic animal’s behaviour changes and thus support the development of new analgesic and neuroactive drugs.
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Young, Suzanne R. L., Christine Doyle, James Marrs, and David G. Skalnik. "CpG Binding Protein Is Required for Hematopoiesis in Zebrafish." Blood 104, no. 11 (November 16, 2004): 3552. http://dx.doi.org/10.1182/blood.v104.11.3552.3552.
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Abstract CpG binding protein (CGBP) is a transcriptional activator that binds unmethylated CpG motifs. CGBP is ubiquitously expressed, and deletion of the CGBP gene in mice results in an embryonic lethal phenotype (E3.5–E6) indicating that CGBP is required for embryonic mammalian development. CGBP-null murine embryonic stem (ES) cells are viable but unable to differentiate and have a 60% decrease in CpG methylation compared to wild type ES cells. Based upon these results, it is hypothesized that CGBP is required for normal stem cell function. RT-PCR was used to identify a homologue of CGBP in zebrafish which is ~70% identical to the mouse CGBP. It was determined that CGBP is expressed in the zebrafish as early as 2 hpf. Morpholino oligonucleotides were designed to the zebrafish CGBP and injected into 1–2 cell embryos. Approximately 80% of the zebrafish treated with the CGBP morpholino oligonucleotides had little or no circulating red blood cells and had abnormal yolk sac morphology at 48 hpf. More than half of the CGBP treated zebrafish also exhibited cardiac edema, and ~14% were dead at 24 hpf. A similar phenotype was observed with a second morpholino designed to an independent site of the zebrafish CGBP cDNA. Zebrafish treated with a control morpholino oligonucleotide at the 1–2 cell stage exhibited normal yolk sac morphology and normal red blood cell circulation at 48 hpf. The control treated zebrafish did not have cardiac edema, and only 2–3% of these fish were dead at 24 hpf. The specificity of phenotypes observed following treatment with the CGBP morpholino oligonucleotide was assessed by co-injection of mouse CGBP mRNA with the CGBP morpholino oligonucleotide at the 1–2 cell stage. Only 37% of these fish exhibited a decrease in circulating red blood cells compared to 80% of zebrafish treated with CGBP morpholino alone. While 14% of the CGBP morpholino treated zebrafish were dead at 24 hpf, only 4% of the co-injected zebrafish were dead at 24 hpf. This experiment demonstrates that the mouse CGBP is able to rescue the phenotypes induced by a reduction of the endogenous zebrafish CGBP. These data indicate that CGBP is essential for normal zebrafish hematopoiesis and establishes the importance of CGBP in post-gastrulation development.
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Nowik, N., P. Podlasz, A. Jakimiuk, N. Kasica, W. Sienkiewicz, and J. Kaleczyc. "Zebrafish: an animal model for research in veterinary medicine." Polish Journal of Veterinary Sciences 18, no. 3 (September 1, 2015): 663–74. http://dx.doi.org/10.1515/pjvs-2015-0086.
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Abstract The zebrafish (Danio rerio) has become known as an excellent model organism for studies of vertebrate biology, vertebrate genetics, embryonal development, diseases and drug screening. Nevertheless, there is still lack of detailed reports about usage of the zebrafish as a model in veterinary medicine. Comparing to other vertebrates, they can lay hundreds of eggs at weekly intervals, externally fertilized zebrafish embryos are accessible to observation and manipulation at all stages of their development, which makes possible to simplify the research techniques such as fate mapping, fluorescent tracer time-lapse lineage analysis and single cell transplantation. Although zebrafish are only 2.5 cm long, they are easy to maintain. Intraperitoneal and intracerebroventricular injections, blood sampling and measurement of food intake are possible to be carry out in adult zebrafish. Danio rerio is a useful animal model for neurobiology, developmental biology, drug research, virology, microbiology and genetics. A lot of diseases, for which the zebrafish is a perfect model organism, affect aquatic animals. For a part of them, like those caused by Mycobacterium marinum or Pseudoloma neutrophila, Danio rerio is a natural host, but the zebrafish is also susceptible to the most of fish diseases including Itch, Spring viraemia of carp and Infectious spleen and kidney necrosis. The zebrafish is commonly used in research of bacterial virulence. The zebrafish embryo allows for rapid, non-invasive and real time analysis of bacterial infections in a vertebrate host. Plenty of common pathogens can be examined using zebrafish model: Streptococcus iniae, Vibrio anguillarum or Listeria monocytogenes. The steps are taken to use the zebrafish also in fungal research, especially that dealing with Candida albicans and Cryptococcus neoformans. Although, the zebrafish is used commonly as an animal model to study diseases caused by external agents, it is also useful in studies of metabolic disorders including fatty liver disease and diabetes. The zebrafish is also a valuable tool as a model in behavioral studies connected with feeding, predator evasion, habituation and memory or lateralized control of behavior. The aim of the present article is to familiarize the reader with the possibilities of Danio rerio as an experimental model for veterinary medicine.
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Jørgensen, Louise von Gersdorff. "Zebrafish as a Model for Fish Diseases in Aquaculture." Pathogens 9, no. 8 (July 27, 2020): 609. http://dx.doi.org/10.3390/pathogens9080609.
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The use of zebrafish as a model for human conditions is widely recognized. Within the last couple of decades, the zebrafish has furthermore increasingly been utilized as a model for diseases in aquacultured fish species. The unique tools available in zebrafish present advantages compared to other animal models and unprecedented in vivo imaging and the use of transgenic zebrafish lines have contributed with novel knowledge to this field. In this review, investigations conducted in zebrafish on economically important diseases in aquacultured fish species are included. Studies are summarized on bacterial, viral and parasitic diseases and described in relation to prophylactic approaches, immunology and infection biology. Considerable attention has been assigned to innate and adaptive immunological responses. Finally, advantages and drawbacks of using the zebrafish as a model for aquacultured fish species are discussed.
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Tsachaki, Maria, Arne Meyer, Benjamin Weger, Denise V. Kratschmar, Janina Tokarz, Jerzy Adamski, Heinz-Georg Belting, Markus Affolter, Thomas Dickmeis, and Alex Odermatt. "Absence of 11-keto reduction of cortisone and 11-ketotestosterone in the model organism zebrafish." Journal of Endocrinology 232, no. 2 (February 2017): 323–35. http://dx.doi.org/10.1530/joe-16-0495.
Full textAbstract:
Zebrafish are widely used as model organism. Their suitability for endocrine studies, drug screening and toxicity assessements depends on the extent of conservation of specific genes and biochemical pathways between zebrafish and human. Glucocorticoids consist of inactive 11-keto (cortisone and 11-dehydrocorticosterone) and active 11β-hydroxyl forms (cortisol and corticosterone). In mammals, two 11β-hydroxysteroid dehydrogenases (11β-HSD1 and 11β-HSD2) interconvert active and inactive glucocorticoids, allowing tissue-specific regulation of glucocorticoid action. Furthermore, 11β-HSDs are involved in the metabolism of 11-oxy androgens. As zebrafish and other teleost fish lack a direct homologue of 11β-HSD1, we investigated whether they can reduce 11-ketosteroids. We compared glucocorticoid and androgen metabolism between human and zebrafish using recombinant enzymes, microsomal preparations and zebrafish larvae. Our results provide strong evidence for the absence of 11-ketosteroid reduction in zebrafish. Neither human 11β-HSD3 nor the two zebrafish 11β-HSD3 homologues, previously hypothesized to reduce 11-ketosteroids, converted cortisone and 11-ketotestosterone (11KT) to their 11β-hydroxyl forms. Furthermore, zebrafish microsomes were unable to reduce 11-ketosteroids, and exposure of larvae to cortisone or the synthetic analogue prednisone did not affect glucocorticoid-dependent gene expression. Additionally, a dual-role of 11β-HSD2 by inactivating glucocorticoids and generating the main fish androgen 11KT was supported. Thus, due to the lack of 11-ketosteroid reduction, zebrafish and other teleost fish exhibit a limited tissue-specific regulation of glucocorticoid action, and their androgen production pathway is characterized by sustained 11KT production. These findings are of particular significance when using zebrafish as a model to study endocrine functions, stress responses and effects of pharmaceuticals.
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YABU, Takeshi, Shuji KISHI, Toshiro OKAZAKI, and Michiaki YAMASHITA. "Characterization of zebrafish caspase-3 and induction of apoptosis through ceramide generation in fish fathead minnow tailbud cells and zebrafish embryo." Biochemical Journal 360, no. 1 (November 8, 2001): 39–47. http://dx.doi.org/10.1042/bj3600039.
Full textAbstract:
Caspase-3 was cloned from zebrafish embryos and its properties were characterized to identify the biological implications of caspase in embryogenesis and apoptosis in zebrafish, which is a model organism in vertebrate developmental biology and genetics. The predicted amino acid sequence, totalling 282 amino acid residues, consisted of the prodomain and large and small subunits. Phylogenetic analysis showed that the cloned zebrafish caspase was a member of the caspase-3 subfamily with approx. 60% identity with caspase-3 from Xenopus, chicken and mammals. In addition, recombinant zebrafish caspase hydrolysed acetyl-Asp-Glu-Val-Asp-4-methyl-coumaryl-7-amide, and exhibited similar substrate specificity to the mammalian caspase-3 subfamily. Therefore this caspase was designated zebrafish caspase-3. Overexpression of zebrafish caspase-3 induced apoptosis and increased ceramide levels in fish fathead minnow tailbud cells and zebrafish embryos. Both ceramide generation and apoptosis induction were inhibited by treatment with a caspase inhibitor, benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone. Moreover, zebrafish caspase-3 mRNA was present in early embryos up to the 1000-cell stage as a maternal factor, and was then expressed throughout the body after the gastrula stage by zygotic expression. These findings indicate that the isolated caspase-3 plays an important role in the induction of ceramide generation as well as apoptosis in fish cells and the zebrafish embryo, and suggest that caspase-3 functions as a modulator of the pro-apoptotic signal in development.
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Swaim, Laura E., Lynn E. Connolly, Hannah E. Volkman, Olivier Humbert, Donald E. Born, and Lalita Ramakrishnan. "Mycobacterium marinum Infection of Adult Zebrafish Causes Caseating Granulomatous Tuberculosis and Is Moderated by Adaptive Immunity." Infection and Immunity 74, no. 11 (November 2006): 6108–17. http://dx.doi.org/10.1128/iai.00887-06.
Full textAbstract:
ABSTRACT The zebrafish, a genetically tractable model vertebrate, is naturally susceptible to tuberculosis caused by Mycobacterium marinum, a close genetic relative of the causative agent of human tuberculosis, Mycobacterium tuberculosis. We previously developed a zebrafish embryo-M. marinum infection model to study host-pathogen interactions in the context of innate immunity. Here, we have constructed a flowthrough fish facility for the large-scale longitudinal study of M. marinum-induced tuberculosis in adult zebrafish where both innate and adaptive immunity are operant. We find that zebrafish are exquisitely susceptible to M. marinum strain M. Intraperitoneal injection of five organisms produces persistent granulomatous tuberculosis, while the injection of ∼9,000 organisms leads to acute, fulminant disease. Bacterial burden, extent of disease, pathology, and host mortality progress in a time- and dose-dependent fashion. Zebrafish tuberculous granulomas undergo caseous necrosis, similar to human tuberculous granulomas. In contrast to mammalian tuberculous granulomas, zebrafish lesions contain few lymphocytes, calling into question the role of adaptive immunity in fish tuberculosis. However, like rag1 mutant mice infected with M. tuberculosis, we find that rag1 mutant zebrafish are hypersusceptible to M. marinum infection, demonstrating that the control of fish tuberculosis is dependent on adaptive immunity. We confirm the previous finding that M. marinum ΔRD1 mutants are attenuated in adult zebrafish and extend this finding to show that ΔRD1 predominantly produces nonnecrotizing, loose macrophage aggregates. This observation suggests that the macrophage aggregation defect associated with ΔRD1 attenuation in zebrafish embryos is ongoing during adult infection.