Zebrafish danio rerio is a vertebrate model system widely used for studies of development and disease modeling. Clonal analysis of cell lineage patterns in plant development. Immersion vaccines are used for a variety of aquacultured fish to protect against infectious diseases caused by bacteria and viruses. Primary culture refers to the stage of the culture after the cells are isolated from the tissue and proliferated under the appropriate conditions until they occupy all of the available substrate i. Stachura dl1, svoboda o, lau rp, balla km, zon li, bartunek p, traver d. Clonal analysis of hematopoietic progenitor cells in the zebrafish article pdf available in blood 1185.
The most interesting of these pathogens is perhaps mycobacterium marinum, a close relative of mycobacterium tuberculosis, and causes fish tuberculosis. Recently, numerous transgenic and mutation technologies have been developed for zebrafish. A clonal analysis of spinal cord development in the zebrafish. Fluorescent in situ hybridization fish is a common inmunohistochemical method used to examine the distribution of rnas in tissue samples. Generation of clonal zebrafish lines and transplantable. Drosophila melanogaster, one of the most powerful genetic models, has allowed the precise manipulation of gene function in specific cell. With the increase in the laboratory use of zebrafish, several advantages, such as a high genetic homology to humans and transparent embryos, which allow clear disease. Mosaic analysis has been used to study the clonal basis of the development of the pigmented retina of the zebrafish, brachydanio rerio.
If not fully anesthetized by time of melanocyte contraction, add a. As vertebrate embryos grow and develop into adults, their organs must acquire mass and mature tissue architecture to maintain proper. Generation of clonal zebrafish line by androgenesis. Genetics, genomics, and transcriptomics, volume 5 4th edition. Ex vivo tools for the clonal analysis of zebrafish hematopoiesis article pdf available in nature protocol 115. Zebrafish are a teleost fish that is part of the cyprinidae family, which includes carps, true minnows, barbs and related genera. Wholeorganism clone tracing using singlecell sequencing nature. Clonal analysis of hematopoietic progenitor cells in the zebra. The zebrafish danio rerio is a freshwater fish belonging to the minnow family of the order cypriniformes. The lack of classical secondary responses following repeated immersion vaccination may. Introduction to cell culture thermo fisher scientific us. To overcome this problem, we generated two lines of homozygous diploid clonal zebrafish lines i. Ongoing genetic screens have identified over 7,000 mutations in zebrafish affecting genes that regulate developmental patterning, organogenesis, physiology and behavior.
Here, we sought to develop a novel method for generation of clonal zebrafish by. Numerous and frequentlyupdated resource results are available from this search. They will use the genetic data from the colourful animal to help them decipher the functions of human genes. The virtually transparent embryos of this species, and the ability to accelerate genetic studies by gene knockdown or overexpression, have led to the widespread use of zebrafish in the detailed investigation of vertebrate gene function and increasingly, the study of human genetic disease 35. We used the zebrafish as a model animal host to investigate how a bacterium, aeromonas veronii. Zebrafish as model organism for drug discovery and. Clonal analysis of hematopoietic progenitor cells in the zebrafish. Depending on the experiment, quantitative analysis of activatedcaspase 3 may require further manipulation of embryos, such as flatmounting, to obtain proper orientation for optimal tissue analysis. The zebrafish, one of the favourite animals of developmental biologists, is rapidly gaining ground in infection models. To investigate the nature of myeloerythroid progenitor. The original zebrafish or zebra danio, danio rerio is a native of rivers in india and bangladesh. This protocol describes the cultivation and differentiation of zebrafish hematopoietic stem and progenitor cells ex vivo for use in colony assays. Ex vivo tools for the clonal analysis of zebrafish.
Pdf ex vivo tools for the clonal analysis of zebrafish. A clonal analysis of segment development in oncopeltus. Clonal and lineage analysis of melanocyte stem cells and their progeny in the zebrafish. In this chapter, through classical behavioural traits, such as swimming capacities, foraging, social. Effects of lethal irradiation in zebrafish and rescue by.
Single cell dynamics of embryonic muscle progenitor cells in zebrafish. We appreciate as much detail as possible and references as appropriate. Various experimental infection models have been set up to study both human and fish pathogens in more detail. Transplantable zebrafish tumors are a novel and very promising model in cancer research. Clonal analysis designates the follow up of the fate or the developmental potential of single cells using either in vivo or cell culture techniques. However, further progress in this field has been contained by a lack of true inbred lines in zebrafish. A multiplex fluorescent in situ hybridization protocol for. Rapid transgenesis techniques, usually based on a tol2 transposon system 1, have been combined with improved cloning options for multiple dna fragment assembly 2. 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. Experimental bacterial adaptation to the zebrafish gut. Genedx 207 perry parkway gaithersburg, md 20877 toll free. 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. Melanocytes are visible in the transparent embryo and in the mature fish following metamorphosis, a.
Zebrafish represent a vertebrate model organism that has been widely, and increasingly, employed over the last decade in the study of developmental processes, wound healing, microbehost interactions, and drug screening. Oclcs webjunction has pulled together information and resources to assist library staff as they consider how to handle coronavirus. Reliable information about the coronavirus covid19 is available from the world health organization current situation, international travel. The data were obtained from embryos in which fluorescent lineage tracer was injected into single cells in the neural plate at the twosomite stage. We describe the results of a clonal analysis of spinal cord development in the zebrafish. Although this is a good basic book on breeding freshwater tropical fish, another book with the same title, from the same publisher written by jay hemdal me. Extraction of proteins from zebrafish embryos for sdsgel analysis western blots of. Author summary animals live in a world teeming with microbes. Zebrafish as model organism for drug discovery and toxicity testing. Such an analysis, however, has not yet been achieved for continuously growing adult tissues, but is essential if we wish to understand the architecture of adult organs. Clonal analysis of segment development 685 table 1. Clonal analysis, the technique of marking somatic cells with mutations uncovered by ionizing radiation, circumvents many of these problems because it identifies clonal cell lini received for publication 21 october 1986. Please use this form to recommend updates to the information in zfin. Power tools for gene expression and clonal analysis in.
The study of melanocyte biology in the zebrafish presents a highly tractable system for understanding fundamental principles of developmental biology. Fish cytogenetics stanford university school of medicine. Cichlids exhibit tremendous diversity in features, such as jaws, teeth and. Adult zebra fish can be anesthetized in a onehalf strength tricaine solution 0.
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. We analyzed clone size and composition in 2 to 3monthold fish. Identification of hematopoietic progenitor cells in the zebrafish danio rerio has been hindered by a lack of functional assays to gauge proliferative potential and differentiation capacity. The zebras 2018 distracted driving report reveals that for the first time since the advent of cell phones, car insurance companies are penalizing drivers who text or use their cell phones while driving. However, there are some differences, in particular in the juvenile and maturationrelated regeneration. Approximately 2 weeks before the procedure, place 10 male and 10 female 612monthold zebrafish into separate standard 9l tanks at. Rapid and efficient zebrafish genotyping using pcr with. Powerful techniques allow efficient generation and recovery of zebrafish mutations. We know very little about the selection pressures that contribute to a microbes ability to be host associated i.
Fish is a molecular cytogenetic technique that can detect chromosomal abnormalities that cannot be appreciated by standard chromosomal analysis e. Water, a motif in the film, best exhibited the symbolism of the fish. We focused on the trunk region of zebrafish that includes the pigment cells. Uk scientists who decoded a third of the human genome say their next target will be the zebrafish. Examined here are crucial topics of juvenility, maturation and rejuvenation in clonal propagation of. Overview zebrafish danio rerio has emerged recently as a premiere organism to study vertebrate development and genetics.
In fact, the average insurance penalty is up nearly 8,000% since 2011 and its costing some people hundreds or even thousands of dollars on their annual premium. The critical comments of margaret poethig and abdollah bassiri are gratefully acknowledged. Cobalt zebra cobalt blue mbuna fish species cobalt zebra picture cool free educational resource gallery jpeg photo representations photograph cobalt zebra photo information info size tank conditions popular name feeding diet description fins gills color behaviour habitat range temperament print illustration pic vivid striking. Clonal analysis of stem cells in differentiation and. It measures three centimeters long and has gold and dark blue stripes. This material is from the 4th edition of the zebrafish book. A clonal analysis of zebrafish heart morphogenesis and regeneration. Injected animals were allowed to survive until either 4 days or 2 weeks postfertilization. Thaker department of veterinary pharmacology and toxicology, college of veterinary science and animal husbandry. Clonal and lineage analysis of melanocyte stem cells and. Classification clones of not restricted to one quadrant of the abdomen class i 3 or all 4 quadrants of abdomen class ii 2 dorsal or 2 ventral quadrants class iii 2 left or 2 right quadrants class iv other combinations of 2 quadrants 06 3 2 2 0 68 3 2 14 0 age at irradiation hours a. Clonal analysis of hematopoietic progenitor cells in the. Briefly, metaphase chromosomes or interphase nuclei are denatured. The morphology of zebrafish colonies generated in these clonal assays closely resembles those generated by mouse and human progenitors, showing that colony shape and size is well conserved among zebrafish and mammals.
During immersion vaccination the antigens are taken up by the skin, gills or gut and processed by the immune system, where the resulting response may lead to protection. Multiple times through the movie, it was mentioned how heavily water impacted edwards life. The big fish was the major symbol that reoccurred multiple times throughout the movie. By the time fish are anesthetized melanocytes are typically contracted. Transplantable tumor lines generated in clonal zebrafish. Effects of domestication on fish behaviour intechopen. Clonal origins of cells in the pigmented retina of the zebrafish eye. New discoveries in the field of stem cell research have frequently appeared in the news and in scientific literature.
In recent nextgeneration sequencing analysis, dhs are helpful for. Clonal analysis is helping us understand the dynamics of cell replacement in homeostatic adult tissues simons and clevers, 2011. Clonal analysis to resolve the fate of individual rather than bulk populations of stem cells has revealed the cellular mechanisms by which stem cells sustain a variety of lineages throughout life. The proliferative diversity between tissues and the dynamic and adaptable nature of the cells that sustain them makes defining the term stem cell. The development of twocomponent expression systems in. In that context, behavioural traits may be affected by human selection. Generation of clonal zebrafish will facilitate largescale genetic screening and help us to overcome other biological and biotechnological challenges due. Generation of clonal zebrafish line by androgenesis without egg. A fresh look at zebrafish from the perspective of cancer. In mosaic tissues composed of a mixed population of wildtype and lossof or gainoffunction mutant cells, fish allows comparison of the effect of the perturbation on gene expression patterns in a mutant. Pigment cell progenitors in zebrafish remain multipotent through. Melanocytes are visible in the transparent embryo and in the mature fish following metamorphosis, a physical transformation from the larval to adult form. In addition, the linearity of the analysis will depend on the thickness of the tissue being analyzed.
Zebrafish have become a popular organism for the study of vertebrate gene function 1,2. To investigate the nature of myeloerythroid progenitor cells, we developed clonal methylcellulose assays by using recombinant zebrafish erythropoietin and granulocyte colonystimulating factor. Domestication is a process by which humans select some phenotypes of wild animal species i. A clonal analysis of zebrafish heart morphogenesis and. Production of homozygous clonal lines of zebrafish. Analysis of apoptosis in zebrafish embryos by wholemount. Differential gene expression analysis in myeloid cells revealed that subpopulation 4. 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. There are several methods for obtaining genomic dna from tail clips provided in the zebrafish book. Research in this area promises to lead to new therapies for cancer, heart disease, diabetes, and a wide variety of other diseases. Generation of clonal zebrafish will facilitate largescale genetic. Basic techniques in genetics and biotechnology of other organisms are generally applicable to forest trees. A guide for the laboratory use of zebrafish danio brachydanio rerio.
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