Analysis of hematopoietic development in the zebra fish book

Recent advances in optical imaging techniques has facilitated tracing of the developmental dynamics of a specific organ, tissue, or even a single cell. Analysis of clonal methylcellulose assays from ism1 morphants also showed. 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. Developing a competitive hematopoietic repopulating assay. 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. Please use this form to recommend updates to the information in zfin. 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. The most interesting of these pathogens is perhaps mycobacterium marinum, a close relative of mycobacterium tuberculosis, and causes fish tuberculosis. Developing a competitive hematopoietic repopulating assay in. These data suggest compensatory regulation of the innate immune system in rag2e450fs mutant zebrafish. Zebrafish has many advantages as a model of human pediatric research.

Zebrafish caudal haematopoietic embryonic stromal tissue. 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. The zebrafish granulocyte colonystimulating factors. Pdf functional analysis of human hematopoietic stem cell gene. Analysis of hematopoietic development during human embryonic ontogenesis. 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.

Flow cytometry data were analyzed using flowjo software version 7. The zebrafish lysozyme c promoter drives myeloidspecific. Zebrafish as a model for leukemia and other hematopoietic. 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. 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. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india, pakistan and other places in asia. Use of the zebrafish danio rerio to define hematopoiesis bahary. Embryonic zebrafish are easily visualized and manipulated through microinjection, chemical treatment, and.

Clearly, if the zebrafish system is to advance the understanding of hematopoiesis at the molecular level, the mutant genes must be isolated. Zebrafish as a model for normal and malignant hematopoiesis. Zebrafish is an ideal vertebrate model system to conduct. 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.

In vivo imaging of hematopoietic stem cell development in the. 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. 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. 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. Methods for assessing drug safety and toxicity wiley. 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. Identification and characterization of these elements should help elucidating transcription regulatory mechanisms of gata2 expression in. The hematopoietic system is in constant contact with stromal cells and endothelial cells during development and in the adult. Involved in several processes, including definitive hemopoiesis. Isthmin 1 ism1 is required for normal hematopoiesis in developing. The zebrafish provides distinct advantages over other vertebrate models during early embryonic development by producing transparent, externally fertilized embryos. A few studies on hematologic toxicology have been undertaken using zebrafish.

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. Recently, several novel regulators of hsc fate decisions have. To facilitate comparison with the lysc transgenic reporter lines generated in this work, we have included a detailed expression analysis of lysc during. In zebrafish, like other vertebrates, hematopoietic development has been. Pdf clonal analysis of hematopoietic progenitor cells in. 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. In vivo imaging is crucial for developmental biology and can further help to follow cell development differentiation in normal and pathological conditions. However, the epigenetic regulation of haematopoiesis regulation via micrornas mirnas remains partially understood. Enrichment of hematopoietic stemprogenitor cells in the zebrafish. Although a substantial number of gene alterations have been identified, the mechanism by which these abnormalities interact has yet to be elucidated.

Because its introduction as a modern experimental model organism. Various experimental infection models have been set up to study both human and fish pathogens in more detail. 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. Use of the zebrafish system to study primitive and definitive. 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. Fluorescenceactivated cell sorting and gene expression analysis ucb or adult bm cd34. Over recent years, zebrafish has been increasingly utilized as a model for a range of human hematopoietic diseases, including malignancies. The zebrafish information network zfin is the database of genetic and genomic data for the zebrafish danio rerio as a model organism. Hematopoietic stem cell development how divergent are. Use of embyronic stem cell in vitro differentiation in a serumfree environment for analysis of mesoderm formation and hematopoietic development.

The zebrafish danio rerio has emerged as a powerful vertebrate genetic and developmental model that is particularly amenable to the study of hematopoiesis. Part vi examines the genetics of hematopoisis, particularly in animal models. The expression pattern of hematopoietic factors during zebra. To investigate the nature of myeloerythroid progenitor cells, we developed clonal methylcellulose assays by using recombinant zebrafish erythropoietin and. Here we identify zebrafish hemogen and show that it is considerably smaller. 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. The bosom map of zebra fish can be evaluate visually hence it is peculiarly susceptible for the perusal of cardiac maps.

Functional comparison of human and zebra fish fkbp52 confirms. Use of zebrafish in drug discovery toxicology chemical. 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. Putative hematopoieticsupportive factors expressed in zebrafish cell. Divergent hemogen genes of teleosts and mammals share. Clonal analysis of hematopoietic progenitor cells in the. In vivo imaging of hematopoietic stem cell development in. The zebrafish granulocyte colonystimulating factors gcsfs. 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.

Hematopoiesis is defined as the formation of all types of blood cells from hematopoietic stem cells hscs orkin and zon, 2008. The hematopoietic system can be assayed in many ways. The zebrafish, one of the favourite animals of developmental biologists, is rapidly gaining ground in infection models. These chapters are meant to be illustrative of the different cell fates, but are not exhaustive.

Research in this area promises to lead to new therapies for cancer, heart disease, diabetes, and a wide variety of other diseases. For example, zebrafish hscs reside in the kidney marrow, which is functionally. In addition, the number of zebra fish cell lines available for in vitro experiments is limited. Analysis of hematopoietic development in the zebrafish. Myeloid malignancies are heterogeneous disorders characterized by uncontrolled proliferation orand blockage of differentiation of myeloid progenitor cells. Dissecting hematopoiesis and disease using the zebrafish. Blood stem cells emerge from aortic endothelium by a novel type of cell transition. 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. Clonal analysis of hematopoietic progenitor cells in the zebra. Thus, our purification method of hscs in the zebrafish kidney is useful to.

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. Zebra fish danio rerio have turn up to be the outstanding theoretical account being in the survey of craniate biological science. B crucial genes during different stages of hematopoietic development in zebrafish. A hematopoiesis in zebrafish occurs in consecutive waves. 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. Interestingly, we detected abnormal cardiac phenotypes and inadequacy of somitegenesis in kdmorphants. Recently, zebrafish has sparked interest in other branches of the biomedical research. Zebrafish as a model for the study of human myeloid. Myelopoiesis in the zebrafish, danio rerio blood american.

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. Cellular and developmental biology, part b, volume 4 3rd edition. The zebrafish model is rapidly gaining prominence in the study of development, hematopoiesis, and disease. In order to isolate novel mutants with hematopoietic defects, largescale mutagenesis screening of zebrafish was performed. Functional analysis of human hematopoietic stem cell gene. Changes in lymphoid progenitor populations during mammalian development. Forward genetic screens are relatively easy, again because of the large number of progeny derived and the speed with which they. We account that knockdown of mir1423p, a hematopoietic specific mirna, in zebrafish occasioned in damage of hematopoiesis during embryonic development.

Use of the zebrafish danio rerio to define hematopoiesis. By scoring specific hematopoietic markers, 52 mutants were identified and then classified into four types based on specific phenotypic traits. A highly conserved regulatory element controls hematopoietic. Regulation of stem cells in the zebra fish hematopoietic system. 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. Development of human lymphohematopoietic stem and progenitor cells defined by expression of cd34 and cd81. The zebrafish is an excellent model for imaging of hematopoiesis due to its transparent.

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. Zebrafish a model system biology essay freebooksummary. These elements, including erythrocytes, granulocytes, monocytes, lymphocytes, and thrombocytes, are formed through complex genetic signaling pathways that are highly conserved throughout phylogeny. The identification of fluorescent zebrafish hscs, coupled with our ability. 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. We appreciate as much detail as possible and references as appropriate. Zfin provides a wide array of expertly curated, organized and crossreferenced zebrafish research data. This unique combination provides the scale, visualization. The zebrafish, danio rerio, is a fast expanding and extremely valuable model system for studying vertebrate developmental biology. Thus, we used a crossspecies comparative approach to assess the residues that are functionally critical for fkbp52 function. 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. Zebrafish are particularly amenable to multigenetic analysis through a. This twovolume reference integrates this exciting area of biology, combining the prerequisites for a general understanding of adult and embryonic.

New discoveries in the field of stem cell research have frequently appeared in the news and in scientific literature. Regulation of stem cells in the zebra fish hematopoietic. Novel insights into the genetic controls of primitive and. Granulocyte colonystimulating factor gcsf drives the proliferation and differentiation of granulocytes, monocytes, and macrophages m. Zebrafish as a developmental model organism for pediatric. Part of the methods in molecular medicine book series mimm, volume 105 log in to check access. Coexpression of lmo2 and gata1 defines a novel hematopoietic population in the pbi. Genetic analysis makes it possible to identify novel genes and, hopefully, genetic pathways that are required for blood formation. Importantly, zebrafish have a strong degree of evolutionary conservation with mammalian hematopoietic development and homeostasis. 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. Mutants of zebra fish effects the peculiar parts of cardiac development. Use of the zebrafish system to study primitive and.

Given the physical and ethical problems with performing experiments on. Early stage zebrafish embryos are particularly suitable for the investigation of hematopoietic development, because they are. The genetic advantages of zebrafish are often considered the strongest. Evidence suggests that endothelial cells and blood cells may arise from a common progenitor, the hemangioblast.

The identification and characterization of zebrafish hematopoietic. 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 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. Embryonic zebrafish are easily visualized and manipulated through microinjection, chemical treatment, and mutagenesis. The development of mature effector cells from upstream hscs, multipotent, oligopotent, and unipotent precursors has served as a paradigm for tissuereplenishing stem cell systems. Comparative analyses of a158 kbp sequence spanning the zebrafish gata2 locus and approximately 400 kbp sequences of gata2 from human, mouse and. Mir145 regulates hematopoiesis during zebrafish development. These studies exemplify the utility of egfptransgenic zebrafish in the analysis of embryonic hematopoietic development. Thus, gata2 and runx1 play pivotal roles in hematopoietic development, affecting mainly the definitive. Dec 29, 2011 the zebrafish model is rapidly gaining prominence in the study of development, hematopoiesis, and disease. The discovery of novel hematopoietic genes, as well as the cloning of zebrafish homologs of known hematopoietic genes, necessitates the use of efficacious and. It is the favoured carnal theoretical account systems in the survey of genetic sciences, toxicology, pharmacological medicine, embryogenesis and cistron map.

Hematopoietic stem cells, hematopoiesis and disease. Zebrafish is a powerful model for the investigation of hematopoiesis. Understanding the genetic controls of hematopoiesis is crucial as perturbations in hematopoiesis lead to diseases such as anemia, thrombocytopenia, or cancers. Analysis of hematopoietic development in the zebrafish springer. 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.

It has been shown that specific expression of gata2 in blood progenitor cells requires distal cisacting regulatory elements. Transgenic zebra fish embryo model for hematopoiesis and. Furthermore, rapid development shows, breaks in less than three days and become mature in 90 days, which makes the search more rapid. 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.

This can confound the analysis of a mutation because more than one. 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 discovery of novel hematopoietic genes, as well as the cloning of zebrafish homologs of known. Mar 16, 2017 importantly, zebrafish have a strong degree of evolutionary conservation with mammalian hematopoietic development and homeostasis. Singlecell transcriptional analysis of normal, aberrant, and. Analysis of the zebrafish genome indicates the presence of 2 gcsf ligands, likely resulting from a duplication event in teleost evolution. Singlecell transcriptional analysis of normal, aberrant. 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.

Over the past decades, zebrafish have become an important model organism, especially in. 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. To functionally assess the progenitor capability of normal and mutant zebra. Strategy to generate a zebrafish lysozyme creporter line. Is expressed in several structures, including head. Methods for assessing drug safety and toxicity offers a practical guide for using zebrafish as a tool for toxicology studies. The zebrafish danio rerio is an especially robust vertebrate system for both. Previous studies demonstrated that the 52kda fk506binding protein fkbp52 prolinerich loop is functionally relevant in the regulation of steroid hormone receptor activity. Clearly, zebrafish scientists studying the development of immune systems have much ground to make up. Functional comparison of human and zebra fish fkbp52. Zebrafish as a model system for the study of hematopoiesis. This complex process involves the coordination of intrinsic master blood transcriptional regulators and signaling molecules from the surrounding tissues huang and zon, 2008. Wholeembryo in situ hybridization analysis showing the localization of hematopoietic factors in zebra. 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.

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. Lessons from transgenic zebrafish expressing the green. Hemogen is a vertebrate transcription factor that performs important functions in erythropoiesis and testicular development and may contribute to neoplasia. 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. Zebrafish hematopoiesis genetic mapping genetic screens vasculogenesis positional cloning. We account that knockdown of mir1423p, a hematopoieticspecific mirna, in zebrafish occasioned in damage of hematopoiesis during embryonic development. Although dispensable in zebra fish, this property allows primitive blood mutants to be studied in the fish because early blood defects in most othervertebratesareembryoniclethal. Aug 20, 2007 gata2 is a transcription factor required for hematopoietic stem cell survival as well as for neuronal development in vertebrates. Largescale forward genetic screening analysis of development. Transcriptome analysis of three distinct hematopoietic cell. 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.