Zebrafish will serve as a onestop resource not only for traditional fish physiologists but also for biomedical researchers, toxicologists and developmental biologists. Native to south asia, it is a popular aquarium fish, frequently sold under the trade name zebra danio and thus often called a tropical fish although both tropical and subtropical. In addition, i summarize current knowledge on the function of maternal factors, both before and after zygotic gene activation, in embryonic processes involved in general cellular functions, axis formation, and cell fate specification. A highquality sequence assembly of the zebrafish genome reveals the largest gene set of any vertebrate and provides information on key genomic features, and comparison to the human reference.
Among the model organisms, the zebrafish danio rerio is one of the best leading models to study. Sequencing of the entire genetic makeup of the zebrafish has revealed that 70 per cent of proteincoding human genes are related to genes found in the zebrafish and that 84 per cent of genes known to be associated with human disease have a zebrafish counterpart. Fish were then killed at weekly intervals for 4 weeks. Pdf identification of novel reference genes suitable for. Scientists use a variety of laboratory techniques to investigate the genetic cause of human diseases. All resources generated by the zgc are publicly accessible to the biomedical research community. The team developed a highquality annotated zebrafish genome sequence to compare with the human reference genome. Eef1a had the lowest variation in expression level between samples n 25 and the cq values differed by less than 4 cycles among samples. Toxicity evaluation and biomarker selection with validated.
A reference growth curve for nutritional experiments in. Cytotoxicity and genotoxicity assays with cultures fish cells. Acknowledging materials and services please acknowledge the zebrafish international resource center in all publications resulting from the materials andor services we are providing to you. Jan 04, 2017 experimental conditions are the conditions under which experiments are performed and data are gathered. The zebrafish information network zfin is the database of genetic and genomic data for. Adult zebrafish as a model organism for behavioural. Pdf zebrafish gapdh can be used as a reference gene for. Like other books in the laboratory animal pocket reference series, this guide covers all aspects pertaining to the use of these organisms including their basic biology, humane care and management, husbandry, life support systems, regulatory compliance, technical procedures, veterinary care, and water quality management. A guide for the laboratory use of zebrafish danio rerio. The choice of the reference gene is a crucial element in qpcr analyses and numerous studies have demonstrated the absence of universal reference gene and the necessity to look for the optimal normalizer for each tissue, condition or species gutierrez et al. Pearl river fisheries research institute, chinese academic of fishery sciences, guangzhou 510380, china. Research often utilizes patients cells or tissue samples, but to determine if a mutation in a specific gene can cause a patients symptoms, we often need experimental animal models. Examples of fish that do well with zebra danios, as listed by aquarium fish, are any of the larger.
The main reason was that the 18s rrna gene exhibited a very high transcript abundance. Characterization of housekeeping genes in zebrafish. Tuba1a was the most variable gene and differed by more than 8 cycles between some. Zebrafish as a model vertebrate for investigating chemical. The zebrafish gene collection zgc is an nih initiative that supports the production of cdna libraries, clones and sequences to provide a complete set of fulllength open reading frame sequences and cdna clones of expressed genes for zebrafish. Morpholino antisense technology reduces gene expression. The best road maps to human bodies lie in the bodies of other animals. The subject areas covered in this volume represent key domains of modern physiological research, which include endocrinology, respiration, cardiovascular function, ionic and.
Researchers discovered a gene also found in humans helps zebrafish convert water motion into. As such, zebra danios are compatible with most species of tropical fish, particularly those described as community fish. The wide sharing of techniques, reagents, and fish lines with in the zebrafish research community has facilitated the rapid growth in research using zebrafish, with a dramatic rise in publications using zebrafish in recent years fig. The zebrafish has become one of the most important model organisms to study biological processes within a living body. Worms in the intestinal bulb of adult zebrafish are motile and can be easily seen when the. Identification of novel reference genes suitable for qrtpcr normalization with respect to the zebrafish developmental stage. Identification of novel reference genes suitable for qrtpcr. A mouse hox gene, hox7, was recently isolated that shows a unique pattern of expression compared to the previously described vertebrate hox genes 1,2. Due to their short lifecycles and relatively large clutch sizes, zebrafish are a useful model for genetic studies. Use features like bookmarks, note taking and highlighting while reading the laboratory zebrafish laboratory animal pocket reference. The zebrafish information network zfin is the database of genetic and genomic data for the zebrafish danio rerio as a model organism.
Among these, a couple of reports described the impact of 2,3,7,8tetrachlorodibenzopdioxin tcdd on dna methylation of embryos, larvae, and adult zebrafish 92, 93. The transcripts selected as potential reference genes were all relatively abundant in tilapia oocytes and their mean cq value ranged between 18. A golden fish reveals pigmentation loss in europeans. Here youll find current best sellers in books, new releases in books, deals in books, kindle ebooks, audible audiobooks, and so much more. Please acknowledge the zebrafish international resource center in all publications resulting from the materials andor services we are providing to you. The zebrafish reference genome sequence and its relationship. These are nonhuman animal species which can be used to understand specific biological processes and to obtain informations which can provide an insight into working of other organisms.
Apr 19, 20 zebrafish genome yields significant similarity to human genome. Entire fish were homogenized and dilution plated on selective 7h10. As a vertebrate that has many of the strengths of invertebrate model systems, it offers numerous advantages to researchers interested in many aspects of embryonic development, physiology and disease. Zebrafish genome yields significant similarity to human genome. However, very little work has been done to assess zebrafish as a model species in nutritional studies in aquaculture in order to utilize the methodological toolbox that this species represents.
Recent research has demonstrated the suitability of adult zebrafish to model some aspects of complex behaviour. In the present study, we used our previous rnaseq dataset to identify novel reference genes suitable for gene expression. Environmental epigenetics in zebrafish epigenetics. Animals as model organisms, the silent sentinels, stand watch over the environmental health of the world. Zhiyuan gong and his colleagues at the national university of singapore were working with a gene that encodes the green fluorescent protein gfp, originally extracted from a jellyfish, that naturally produced. The isolation and molecular analysis of zebrafish behavioural mutants is now starting.
Although realtime reverse transcription pcr qpcr, normalized to an internal reference housekeeping gene, is a frequently used method for quantifying gene expression changes in zebrafish, many commonly used housekeeping genes are known to vary with experimental conditions. The zebrafish, a small but flashy aquarium pet, may seem like an unlikely informant on questions of human genetics yet its genome could hold the keys to understanding many diseases and, surprisingly, the genes underlying human skin color. Reference genes to quantify gene expression during. Zebrafish gapdh can be used as a reference gene for expression analysis in crosssubfamily cloned embryos. It was developed by biomedical researchers in the early 1980s to detect and localize the presence or absence of specific dna. Mos have been shown to successfully knockdown gene expression in zebrafish embryos nasevicius and ekker, 2001 by injecting complementary sequences to either the 25 bases 3. Only two other large genomes have been sequenced to this high standard. Each dish was wrapped in parafilm then placed atop a 3inch spacer to achieve a dose rate of 978 cgymin in a 7 cesium source irradiator. This material is from the 4th edition of the zebrafish book. Native to south asia, it is a popular aquarium fish, frequently sold under the trade name zebra danio and thus often called a tropical fish although both tropical and subtropical the zebrafish is an important and widely used vertebrate model organism in scientific research, for.
Shared intronexon structure and gene order slc24a5, myef2, ctxn2, and slc12a5 between fish and mammals further supports the conclusion that the zebrafish golden gene and slc24a5 are orthologs. Zebrafishmycobacterium marinum model for mycobacterial. A few recent studies have suggested that pollutant exposure could induce alteration in dna methylation patterns by disturbing dnmt gene expression during zebrafish embryogenesis. However, many traditional reference genes used in zebrafish early development are not appropriate because of their variable expression levels. It measures three centimeters long and has gold and dark blue stripes. Nov 12, 2008 research using the zebrafish model has experienced a rapid growth in recent years. It was developed by biomedical researchers in the early 1980s to detect and localize the presence or absence of specific dna sequences on chromosomes.
Hardcopies of the 4th edition of the zebrafish book can be obtained for a nominal fee from the zebrafish international resource center, 5274 university of oregon, eugene, or 97403 usa. Fluorescence in situ hybridization fish is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of a nucleic acid sequence with a high degree of sequence complementarity. The laboratory zebrafish crc press book like other books in the laboratory animal pocket reference series, this guide covers all aspects pertaining to the use of these organisms including their basic biology, humane care and management, husbandry, life support systems, regulatory compliance, technical procedures, veterinary care, and wate. However, formatting rules can vary widely between applications and fields of interest or study. Identification of housekeeping genes suitable for gene. The zebrafish book a guide for the laboratory use of zebrafish danio brachydanio rerio by monte westerfield, institute of neuroscience, university of oregon older literature referred to zebrafish as brachydanio rerio. In december 2005, a cancer research team headed by keith cheng at penn state university announced that their studies of the mutant golden zebrafish had. Zfin provides a wide array of expertly curated, organized and crossreferenced zebrafish research data. Zebrafish danio rerio as a model organism intechopen. It turns out that being a paleontologist is a huge advantage in teaching human anatomy.
Studies of reward behaviour, learning and memory, aggression, anxiety and sleep strongly suggest that conserved regulatory processes underlie behaviour in zebrafish and mammals. If this situation were to occur when testing a novel chemical, or when. Identification of novel reference genes suitable for qrt. Wellcome genome campus hinxton, cambridgeshire, cb10 1sa. This book includes subjects relating to zebrafish physiology. Characterization of three zebrafish genes related to hox7. We perform a systematic analysis to identify optimal hk genes in danio rerio zebrafish, to be used in expression analyses conducted on embryoslarvae at. While mice and rats have been common choices for modeling human diseases in the past, the use of zebrafish. Methods for assessing drug safety and toxicity offers a practical guide for using zebrafish as a tool for toxicology studies. However, many traditional reference genes used in zebrafish early development are not appropriate because of their variable expression levels during embryogenesis. Identification of housekeeping genes suitable for gene expression. Pdf characterization of housekeeping genes in zebrafish. Identification of housekeeping genes suitable for gene expression analysis in the zebrafish. Validation of zebrafish danio rerio reference genes for.
Reference genes used in normalizing qrtpcr data are critical for the accuracy of gene expression analysis. A guide for the laboratory use of zebrafish danio rerio, eugene, university of oregon press. The original zebrafish or zebra danio, danio rerio is a native of rivers in india and bangladesh. The role of maternal factors in early zebrafish development. Download it once and read it on your kindle device, pc, phones or tablets. Zebrafish genome yields significant similarity to human. This book not only provides a complete set of instructions that will allow.
Consolidating key protocols and approaches to help researchers navigate the important and evolving field of zebrafish models for toxicity screening, this new title describes the methods for using the zebrafish as a model organism to assess compoundinduced. The laboratory zebrafish laboratory animal pocket reference kindle edition by harper, claudia, lawrence, christian. Zebrafish genome found strikingly similar to humans. The zebrafish danio rerio is a freshwater fish belonging to the minnow family of the order cypriniformes. Reference genes to quantify gene expression during oogenesis. Goulds book of fish is a novel about fish the way mobydick is a novel about whales, or ulysses is a novel about the events of a single day. Zebra danios are peaceful schooling fish tolerant of a wide range of aquarium conditions, according to animal world. Dec 14, 2010 like other books in the laboratory animal pocket reference series, this guide covers all aspects pertaining to the use of these organisms including their basic biology, humane care and management, husbandry, life support systems, regulatory compliance, technical procedures, veterinary care, and water quality management. Early lifestage test in zebra fish versus a growth test in rainbow trout to evaluate toxic effects. The 18s rrna gene was not recommended as a reference gene in the present study, whereas with the least m value in bestkeeper analysis. Apr 17, 20 a highquality sequence assembly of the zebrafish genome reveals the largest gene set of any vertebrate and provides information on key genomic features, and comparison to the human reference. In the relatively brief span of only a few decades. The simplest way to teach students the nerves in the human head is to show them the state of affairs in sharks.
These fish are a valuable tool for examining erythrocyte development and circulation defects in the developing embryo. Methods for assessing drug safety and toxicity wiley. Research using the zebrafish model has experienced a rapid growth in recent years. In december 2005, a cancer research team headed by keith cheng at. Acute irradiation injury was replicated using animal models, and seminal experiments by jacobsen et al 2,3 and lorenz et al 4 showed that shielding or. They can range from standard, as defined in the zebrafish book or generic control nonstandard control treatments such as a vehicle dmso injection to any sort of manipulation of the environment of the fish as in exposure to a chemical or a nonstandard lightdark cycle or. A highquality sequence assembly of the zebrafish genome reveals the largest gene set of any vertebrate and provides information on key genomic features, and comparison to. This cuttingedge resource includes uptodate information on zebrafish physiology and the tools used to study it, not only as a model species for studies of other vertebrates but with application. In this study, to systematically identify the most stably expressed genes for internal reference in zebrafish danio rerio investigations, 37 d. The high sequence similarity among the orthologous sequences from fish and mammals fig. Hox7 is expressed in the cephalic neural crest and its mesectodermal derivatives in the branchial arch region. The earliest events in zebrafish development are driven by maternal factors deposited in the egg during oogenesis that become activated upon fertilization and initiate cascades of events that drive early development.
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