Analysis of hematopoietic development in the zebra fish book

Fluorescenceactivated cell sorting and gene expression analysis ucb or adult bm cd34. The zebrafish granulocyte colonystimulating factors. Use of the zebrafish danio rerio to define hematopoiesis. While genetically distant from humans, nonetheless the vertebrate zebrafish has comparable organs and tissues that make it the model organism for study of vertebrate development. Zebrafish as a model for leukemia and other hematopoietic. Recently, several novel regulators of hsc fate decisions have. Analysis of clonal methylcellulose assays from ism1 morphants also showed. Early indications are that many of the hematopoietic mutations in zebrafish may correspond to novel genes, because few of the blood mutants correspond to the 30 mapped candidate genes leonard zon, unpublished data. Because its introduction as a modern experimental model organism. Please use this form to recommend updates to the information in zfin. We account that knockdown of mir1423p, a hematopoieticspecific mirna, in zebrafish occasioned in damage of hematopoiesis during embryonic development. Changes in lymphoid progenitor populations during mammalian development. For example, zebrafish hscs reside in the kidney marrow, which is functionally. Use of embyronic stem cell in vitro differentiation in a serumfree environment for analysis of mesoderm formation and hematopoietic development.

Interestingly, we detected abnormal cardiac phenotypes and inadequacy of somitegenesis in kdmorphants. Zebrafish is an ideal vertebrate model system to conduct. New discoveries in the field of stem cell research have frequently appeared in the news and in scientific literature. B crucial genes during different stages of hematopoietic development in zebrafish.

Use of the zebrafish danio rerio to define hematopoiesis bahary. Although the kidney is the site of hematopoiesis in the adult fish, the predecessors of the stem cells responsible for definitive hematopoiesis first appear in a region of the trunk called the aortagonadmesonephros agm at about 24 hpf, whence they move to another temporary hematopoietic site in the ventral tail called the caudal. Research in this area promises to lead to new therapies for cancer, heart disease, diabetes, and a wide variety of other diseases. The zebrafish model is rapidly gaining prominence in the study of development, hematopoiesis, and disease. The development of mature effector cells from upstream hscs, multipotent, oligopotent, and unipotent precursors has served as a paradigm for tissuereplenishing stem cell systems. Various experimental infection models have been set up to study both human and fish pathogens in more detail.

The zebrafish danio rerio is an especially robust vertebrate system for both. Zebra fish danio rerio have turn up to be the outstanding theoretical account being in the survey of craniate biological science. In vivo imaging is crucial for developmental biology and can further help to follow cell development differentiation in normal and pathological conditions. Genetic analysis makes it possible to identify novel genes and, hopefully, genetic pathways that are required for blood formation. This complex process involves the coordination of intrinsic master blood transcriptional regulators and signaling molecules from the surrounding tissues huang and zon, 2008. Use of zebrafish in drug discovery toxicology chemical. This twovolume reference integrates this exciting area of biology, combining the prerequisites for a general understanding of adult and embryonic. Functional comparison of human and zebra fish fkbp52 confirms. Using a double transgenic fish that has a gata1 promoter driving dsred and globin lcr driving gfp, the stages of erythroid development can be studied. Regulation of stem cells in the zebra fish hematopoietic.

The genetic advantages of zebrafish are often considered the strongest. Part vi examines the genetics of hematopoisis, particularly in animal models. Dissecting hematopoiesis and disease using the zebrafish. Clearly, zebrafish scientists studying the development of immune systems have much ground to make up. These data suggest compensatory regulation of the innate immune system in rag2e450fs mutant zebrafish. Zebrafish as a model for normal and malignant hematopoiesis. Zfin provides a wide array of expertly curated, organized and crossreferenced zebrafish research data. Hematopoiesis is defined as the formation of all types of blood cells from hematopoietic stem cells hscs orkin and zon, 2008. In vivo imaging of hematopoietic stem cell development in the. Zebrafish a model system biology essay freebooksummary. The hematopoietic system is in constant contact with stromal cells and endothelial cells during development and in the adult. The results of in situ hybridization experiments using zebrafish homologues of 3 early mammalian hematopoietic genes, scl, lmo2, and gata2, indicate analogous expression patterns during zebrafish hematopoiesis. Here, in this study we show that mirna145 participates in the haematopoiesis regulation and vascular development of the early stages of erythropoiesis during zebra fish early developmental stages. The zebrafish danio rerio has emerged as a powerful vertebrate genetic and developmental model that is particularly amenable to the study of hematopoiesis.

Use of the zebrafish system to study primitive and definitive. The zebrafish lysozyme c promoter drives myeloidspecific. Analysis of hematopoietic development during human embryonic ontogenesis. We account that knockdown of mir1423p, a hematopoietic specific mirna, in zebrafish occasioned in damage of hematopoiesis during embryonic development. The discovery of novel hematopoietic genes, as well as the cloning of zebrafish homologs of known. Zebrafish danio rerio is an excellent model of vertebrate embryonic development, since it has many advantages such as its small size, large number of progeny, transparent embryos, and short life cycle amatruda and zon, 1999. A few studies on hematologic toxicology have been undertaken using zebrafish.

Putative hematopoieticsupportive factors expressed in zebrafish cell. The hematopoietic system can be assayed in many ways. The zebrafish has emerged as an ideal organism for the study of hematopoiesis, the process by which all the cellular elements of the blood are formed. This unique combination provides the scale, visualization. To functionally assess the progenitor capability of normal and mutant zebra. Methods for assessing drug safety and toxicity offers a practical guide for using zebrafish as a tool for toxicology studies. Zebrafish has many advantages as a model of human pediatric research. Hematopoiesis is a dynamic process where initiation and maintenance of hematopoietic stem cells, as well as their differentiation into erythroid, myeloid and lymphoid lineages, are tightly regulated by a network of transcription factors. Aug 01, 2001 the results of in situ hybridization experiments using zebrafish homologues of 3 early mammalian hematopoietic genes, scl, lmo2, and gata2, indicate analogous expression patterns during zebrafish hematopoiesis. Early stage zebrafish embryos are particularly suitable for the investigation of hematopoietic development, because they are. Thus, gata2 and runx1 play pivotal roles in hematopoietic development, affecting mainly the definitive. Due to its prolific reproduction and the external development of the transparent embryo, the zebrafish is the prime model for genetic and developmental studies, as well as research in genomics. The developmental processes and molecular mechanisms involved in zebrafish hematopoiesis are conserved with higher vertebrates, and the genetic and experimental accessibility of the fish and the optical transparency of its embryos and larvae make it ideal for in vivo analysis of hematopoietic development. Clearly, if the zebrafish system is to advance the understanding of hematopoiesis at the molecular level, the mutant genes must be isolated.

In zebrafish, like other vertebrates, hematopoietic development has been. The development of clonal in vitro cultures by metcalf and colleagues in the 1960s enabled the growth of murine bone marrow progenitors 1 and the study and quantitation of progenitor number during hematologic disease 2 and exposure to irradiation. Methods for assessing drug safety and toxicity wiley. Functional comparison of human and zebra fish fkbp52. The identification of fluorescent zebrafish hscs, coupled with our ability.

To investigate the nature of myeloerythroid progenitor cells, we developed clonal methylcellulose assays by using recombinant zebrafish erythropoietin and. Part of the methods in molecular medicine book series mimm, volume 105 log in to check access. Blood stem cells emerge from aortic endothelium by a novel type of cell transition. Zebrafish as a model for the study of human myeloid. Granulocyte colonystimulating factor gcsf drives the proliferation and differentiation of granulocytes, monocytes, and macrophages m. Analysis of egfp expression in larvae and adult transgenic zebrafish the major site of definitive hematopoiesis in the zebrafish beginning at 710 dpf and continuing through adulthood lies within the interstitium of the. Mir145 regulates hematopoiesis during zebrafish development. Hematopoietic stem cells, hematopoiesis and disease. The identification and characterization of zebrafish hematopoietic.

The discovery of novel hematopoietic genes, as well as the cloning of zebrafish homologs of known hematopoietic genes, necessitates the use of efficacious and. Zebrafish is a powerful vertebrate system to study myelination using both forward and reverse genetic approaches, in vivo analysis during development and repair, and for performing drug screens to uncover novel drivers of myelination and remyelination. Recently, zebrafish has sparked interest in other branches of the biomedical research. The zebrafish granulocyte colonystimulating factors gcsfs. Evidence suggests that endothelial cells and blood cells may arise from a common progenitor, the hemangioblast. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india, pakistan and other places in asia.

These chapters are meant to be illustrative of the different cell fates, but are not exhaustive. Cellular and developmental biology, part b, volume 4 3rd edition. Here we identify zebrafish hemogen and show that it is considerably smaller. The zebrafish embryo develops externally and its optical clarity allows the number and morphology of circulating blood cells to be visualized using a dissecting microscope. Isthmin 1 ism1 is required for normal hematopoiesis in developing.

This book, one of two new volumes in the reliable lab solutions series dealing with zebrafish, brings together a robust and uptodate collection of timetested. However, with the development of new technologies such as omics approaches at singlecell resolution, recent studies in vitro and in vivo have suggested that heterogeneity is a common. Forward genetic screens are relatively easy, again because of the large number of progeny derived and the speed with which they. Largescale forward genetic screening analysis of development. Although a substantial number of gene alterations have been identified, the mechanism by which these abnormalities interact has yet to be elucidated. In addition, the zebra fish reproduces rapidly a single pair can produce hundreds of embryos per week and mass mating strategies can produce tens of thousands of synchronized offspring.

Analysis of hematopoietic development in the zebrafish. Clonal analysis of hematopoietic progenitor cells in the. Divergent hemogen genes of teleosts and mammals share. These studies exemplify the utility of egfptransgenic zebrafish in the analysis of embryonic hematopoietic development. Embryonic zebrafish are easily visualized and manipulated through microinjection, chemical treatment, and mutagenesis. Identification and characterization of these elements should help elucidating transcription regulatory mechanisms of gata2 expression in. Zebrafish as a developmental model organism for pediatric. Flow cytometry data were analyzed using flowjo software version 7. This can confound the analysis of a mutation because more than one.

Download citation analysis of hematopoietic development in the zebrafish the zebrafish danio rerio has emerged as a powerful vertebrate genetic and developmental model that is particularly. By scoring specific hematopoietic markers, 52 mutants were identified and then classified into four types based on specific phenotypic traits. Myeloid malignancies are heterogeneous disorders characterized by uncontrolled proliferation orand blockage of differentiation of myeloid progenitor cells. The zebrafish information network zfin is the database of genetic and genomic data for the zebrafish danio rerio as a model organism. Developing a competitive hematopoietic repopulating assay in. Over recent years, zebrafish has been increasingly utilized as a model for a range of human hematopoietic diseases, including malignancies. Singlecell transcriptional analysis of normal, aberrant. Over the past decades, zebrafish have become an important model organism, especially in. Recent advances in optical imaging techniques has facilitated tracing of the developmental dynamics of a specific organ, tissue, or even a single cell. Zebrafish is a powerful model for the investigation of hematopoiesis. Dec 29, 2011 the zebrafish model is rapidly gaining prominence in the study of development, hematopoiesis, and disease. Aug 04, 2011 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. In vivo imaging of hematopoietic stem cell development in. It has been shown that specific expression of gata2 in blood progenitor cells requires distal cisacting regulatory elements.

Novel insights into the genetic controls of primitive and. The bosom map of zebra fish can be evaluate visually hence it is peculiarly susceptible for the perusal of cardiac maps. Comparative analyses of a158 kbp sequence spanning the zebrafish gata2 locus and approximately 400 kbp sequences of gata2 from human, mouse and. Transcriptome analysis of three distinct hematopoietic cell. Furthermore, rapid development shows, breaks in less than three days and become mature in 90 days, which makes the search more rapid. Developing a competitive hematopoietic repopulating assay in zebrafish project narrative hematopoietic stem cells hscs residing in the bone marrow normally expand and differentiate to produce all the different types of blood cell types that an organism needs during its lifetime, a tightly regulated process that is poorly understood.

The kidney is the adult hematopoietic organ in zebra fish equivalent to mammalian bone marrow, and likethecht,cmyb,scl,runx1,andikarosareexpressed in this region murayama et al. Development of human lymphohematopoietic stem and progenitor cells defined by expression of cd34 and cd81. Coexpression of lmo2 and gata1 defines a novel hematopoietic population in the pbi. However, the epigenetic regulation of haematopoiesis regulation via micrornas mirnas remains partially understood. Zebrafish as a model system for the study of hematopoiesis. Pdf clonal analysis of hematopoietic progenitor cells in. The expression pattern of hematopoietic factors during zebra. Proliferative capacity of murine hematopoietic stem cells. Lessons from transgenic zebrafish expressing the green.

May 21, 2011 in vivo imaging is crucial for developmental biology and can further help to follow cell development differentiation in normal and pathological conditions. Zebrafish is an established model for the study of vertebrate development, and is especially amenable for investigating hematopoiesis, where there is strong conservation of key lineages, genes, and developmental processes with humans. The zebrafish provides distinct advantages over other vertebrate models during early embryonic development by producing transparent, externally fertilized embryos. Embryonic zebrafish are easily visualized and manipulated through microinjection, chemical treatment, and. Involved in several processes, including definitive hemopoiesis. The lysc promoter region was selected for generation of a reporter line, as expression of the lysc gene had been described as being specific to the zebrafish macrophage lineage. Zebrafish are particularly amenable to multigenetic analysis through a. We use cookies to offer you a better experience, personalize content, tailor advertising, provide social media features, and better understand the use of our services. Previous studies demonstrated that the 52kda fk506binding protein fkbp52 prolinerich loop is functionally relevant in the regulation of steroid hormone receptor activity.

A hematopoiesis in zebrafish occurs in consecutive waves. We appreciate as much detail as possible and references as appropriate. The bosom whipping begins precisely at 22hpf and peristaltic moving ridge at first at 36 hpf for the atrium and ventricle as a coordinate contractions. Strategy to generate a zebrafish lysozyme creporter line. Understanding the genetic controls of hematopoiesis is crucial as perturbations in hematopoiesis lead to diseases such as anemia, thrombocytopenia, or cancers. The zebrafish is an excellent model for imaging of hematopoiesis due to its transparent. In addition, the number of zebra fish cell lines available for in vitro experiments is limited. Zebrafish caudal haematopoietic embryonic stromal tissue. Methods for assessing drug safety and toxicity offers a practical guide for using zebrafish as a tool for toxicology studies 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. In the past two decades, zebrafish has become one of the preferred in vivo model organisms for studying diverse processes like embryogenesis 8, 9, autophagy, behaviour 11, 12 and sleep. The classical roadmap of hematopoietic hierarchy has been proposed for nearly 20 years and has become a dogma of stem cell research for most types of adult stem cells, including hematopoietic stem cells hscs. Hematopoietic stem cell development how divergent are. Functional analysis of human hematopoietic stem cell gene.

It is the favoured carnal theoretical account systems in the survey of genetic sciences, toxicology, pharmacological medicine, embryogenesis and cistron map. The expression patterns of gata2 in hematopoietic and neuronal tissues are conserved in vertebrates, suggesting that the arrangement and sequence of the gata2 genomic locus may be highly conserved. To facilitate comparison with the lysc transgenic reporter lines generated in this work, we have included a detailed expression analysis of lysc during. Importantly, zebrafish have a strong degree of evolutionary conservation with mammalian hematopoietic development and homeostasis. Thus, our purification method of hscs in the zebrafish kidney is useful to. Embryonic primitive hematopoiesis yellow starts at around 11 hours post fertilization hpf when hemangioblasts which have the potential to become either endothelial vascular cells or hematopoietic cells, shown in white appear in the anterior lateral mesoderm alm and posterior lateral. Use of the zebrafish system to study primitive and. Given the physical and ethical problems with performing experiments on. Aug 20, 2007 gata2 is a transcription factor required for hematopoietic stem cell survival as well as for neuronal development in vertebrates. Transgenic zebra fish embryo model for hematopoiesis and. Myelopoiesis in the zebrafish, danio rerio blood american. Analysis of hematopoietic development in the zebrafish springer. Analysis of the zebrafish genome indicates the presence of 2 gcsf ligands, likely resulting from a duplication event in teleost evolution. The hematopoietic phenotypes may result from mutations in either novel genes or in the zebrafish homologs of genes known to be required for hematopoiesis in higher vertebrates.

Mutants of zebra fish effects the peculiar parts of cardiac development. Dr fkbp52 contains all of the analogous domains and residues previously identified as critical for fkbp52 potentiation of receptor activity, it fails to potentiate activity. Singlecell transcriptional analysis of normal, aberrant, and. The zebrafish, danio rerio, is a fast expanding and extremely valuable model system for studying vertebrate developmental biology. Although dispensable in zebra fish, this property allows primitive blood mutants to be studied in the fish because early blood defects in most othervertebratesareembryoniclethal. The zebrafish as a model for hostpathogen interactions has now matured to the point that we can reflect on what it truly has to offer, where it is helpful, and how it complements other models. We and others have successfully shown that dramatically increased hematopoietic development can be modeled in zebrafish, as is the case for knockdown of the bmpantagonist chd chordin and the corresponding dino mutant 41,42. The most interesting of these pathogens is perhaps mycobacterium marinum, a close relative of mycobacterium tuberculosis, and causes fish tuberculosis. Hemogen is a vertebrate transcription factor that performs important functions in erythropoiesis and testicular development and may contribute to neoplasia. 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. Clonal analysis of hematopoietic progenitor cells in the zebra.

Pdf functional analysis of human hematopoietic stem cell gene. The transparency of the zebra fish embryo facilitates the studies in genetic development programs, because it is possible to monitor and manipulate its development without difficulties. Wholeembryo in situ hybridization analysis showing the localization of hematopoietic factors in zebra. Mar 16, 2017 importantly, zebrafish have a strong degree of evolutionary conservation with mammalian hematopoietic development and homeostasis. Regulation of stem cells in the zebra fish hematopoietic system. The zebrafish, one of the favourite animals of developmental biologists, is rapidly gaining ground in infection models. These elements, including erythrocytes, granulocytes, monocytes, lymphocytes, and thrombocytes, are formed through complex genetic signaling pathways that are highly conserved throughout phylogeny. Zebrafish hematopoiesis genetic mapping genetic screens vasculogenesis positional cloning. Thus, we used a crossspecies comparative approach to assess the residues that are functionally critical for fkbp52 function.