The zebrafish spi1 promoter drives myeloid-specific expression in stable transgenic fish

Ward, Alister C.; McPhee, Dora; Condron, Melanie; Varma, Sony; Cody, Stephen H.; Onnebo, Sara M.N.; Paw, Barry H.; Zon, Leonard I.; Lieschke, Graham J.

doi:10.1182/blood-2003-03-0966

Cited by 89 publications

(74 citation statements)

References 23 publications

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“…Interestingly, a clear cell subpopulation structure emerged ( Fig. 2A) showing three distinct cell groups with distinct expression of known immune markers such as cd8 and cd4 for T cells, nitr and dicp for teleost fish NK-like cells , and spi1b for myeloid cells (see Methods) (Ward et al 2003). Next, we performed differential expression analysis of the cells expressing these markers to derive broader zebrafish T-cell, NK-like, and myeloid-like cell signatures (see Methods) (see Supplemental Table S1) and determine whether absence of expression of specific markers in many cells was mainly due to technical limitations of single-cell RNA-seq technology (i.e., high transcript dropout rates).…”

Section: Conservation Analysis Of Mammalian T- B- Nk- and Myeloidcmentioning

confidence: 99%

Single-cell transcriptome analysis of fish immune cells provides insight into the evolution of vertebrate immune cell types

et al. 2017

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The immune system of vertebrate species consists of many different cell types that have distinct functional roles and are subject to different evolutionary pressures. Here, we first analyzed conservation of genes specific for all major immune cell types in human and mouse. Our results revealed higher gene turnover and faster evolution of trans-membrane proteins in NK cells compared with other immune cell types, and especially T cells, but similar conservation of nuclear and cytoplasmic protein coding genes. To validate these findings in a distant vertebrate species, we used single-cell RNA sequencing of lck:GFP cells in zebrafish and obtained the first transcriptome of specific immune cell types in a nonmammalian species. Unsupervised clustering and single-cell TCR locus reconstruction identified three cell populations, T cells, a novel type of NK-like cells, and a smaller population of myeloid-like cells. Differential expression analysis uncovered new immunecell-specific genes, including novel immunoglobulin-like receptors, and neofunctionalization of recently duplicated paralogs. Evolutionary analyses confirmed the higher gene turnover of trans-membrane proteins in NK cells compared with T cells in fish species, suggesting that this is a general property of immune cell types across all vertebrates.

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Section: Conservation Analysis Of Mammalian T- B- Nk- and Myeloidcmentioning

confidence: 99%

Single-cell transcriptome analysis of fish immune cells provides insight into the evolution of vertebrate immune cell types

et al. 2017

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“…The development of transgenic zebrafish lines with fluorescently labeled neutrophils and/or macrophages has revolutionized our ability to dissect the molecular mechanisms that regulate leukocyte motility and trafficking during normal homeostasis and following acute or chronic inflammation (summarized in Table 2) (Lawson and Weinstein, 2002;Ward et al, 2003;Hsu et al, 2004;Mathias et al, 2006;Renshaw et al, 2006;Hall et al, 2007;Mathias et al, 2009b;Feng et al, 2010;Ellett et al, 2011;Gray et al, 2011;Wittamer et al, 2011). During zebrafish development, neutrophils comprise two distinct populations: a relatively stationary population in the caudal hematopoietic tissue, and a spontaneously highly motile population in the rostral mesenchyme .…”

Section: Neutrophil and Macrophage Migration In Zebrafish Embryosmentioning

confidence: 99%

Leukocyte migration from a fish eye's view

Deng

Huttenlocher

2012

Journal of Cell Science

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SummaryIn the last five years, the zebrafish (Danio rerio) has rapidly gained popularity as a model system for studying leukocyte migration and trafficking in vivo. The optical clarity of zebrafish embryos, as well as the potential for genetic manipulation and the development of tools for live imaging, have provided new insight into how leukocytes migrate in response to directional cues in live animals. This Commentary discusses recent progress in our understanding of how leukocytes migrate in vivo, including the role of intracellular signaling through phosphatidylinositol 3-kinase (PI3K) in both random and directed migration. The importance of leukocyte reverse migration in the resolution of inflammation will also be discussed. Finally, we will highlight how zebrafish models have helped to provide new insight into leukocyte migration and the way in which migration is altered in disease.

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“…6,14 These primitive macrophages can be visualized in vivo using a variety of transgenic lines in which green fluorescent protein (GFP) expression is driven by myeloid-specific promoter sequences, including pu.1:eGFP, lyz:eGFP, and mpx:eGFP. [15][16][17][18][19] However, these promoters are only specific to macrophages during a short, defined window of embryogenesis; each is also subsequently expressed in granulocytic cells, precluding the prospective isolation of mononuclear phagocytes after 48 hpf. Although a novel macrophage-specific reporter line was recently generated using the mpeg1 promoter, it appears that the transgene is active only in the embryo and larvae, precluding its use for the study of macrophages in adult fish.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the mononuclear phagocyte system in zebrafish

Wittamer

Gutschow

Traver

2011

Blood

187

180

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The evolutionarily conserved immune system of the zebrafish (Danio rerio), in combination with its genetic tractability, position it as an excellent model system in which to elucidate the origin and function of vertebrate immune cells. We recently reported the existence of antigenpresenting mononuclear phagocytes in zebrafish, namely macrophages and dendritic cells (DCs), but have been impaired in further characterizing the biology of these cells by the lack of a specific transgenic reporter line. Using regulatory elements of a class II major histocompatibility gene, we generated a zebrafish reporter line expressing green fluorescent protein (GFP) in all APCs, macrophages, DCs, and B lymphocytes. Examination of mhc2dab:GFP; cd45:DsRed doubletransgenic animals demonstrated that kidney mhc2dab:GFP hi ; cd45:DsRed hi cells were exclusively mature monocytes/macrophages and DCs, as revealed by morphologic and molecular analyses. Mononuclear phagocytes were found in all hematolymphoid organs, but were most abundant in the intestine and spleen, where they up-regulate the expression of inflammatory cytokines upon bacterial challenge. Finally, mhc2dab:GFP and cd45:DsRed transgenes mark mutually exclusive cell subsets in the lymphoid fraction, enabling the delineation of the major hematopoietic lineages in the adult zebrafish. These findings suggest that mhc2dab:GFP and cd45:DsRed transgenic lines will be instrumental in elucidating the immune response in the zebrafish. (Blood. 2011;117(26):7126-7135) IntroductionIn recent years, the zebrafish (Danio rerio) has proven to be a unique vertebrate model for the study of hematopoiesis. 1 The use of the zebrafish to study the ontogeny of leukocyte subsets, 2 immune cell migration, 3,4 and host-pathogen interactions 5 has provided new insights into our understanding of innate immunity in the developing vertebrate embryo. A major focus of previous studies was on the neutrophil response, because several transgenic reporter lines have been generated that mark this granulocyte subset. Whereas neutrophils generally constitute the first line of defense against invading pathogens, the role of other immune cell subsets in the innate immune response has received less attention. In addition to neutrophils, macrophages are key in the response to pathogen challenge. In the zebrafish embryo, primitive macrophages have been demonstrated to be capable of clearing injected bacteria by phagocytosis. 6 However, the absence of markers specific to macrophages has limited the study of this myeloid cell subset in the zebrafish.The mononuclear phagocyte system (MPS) comprises monocytes, tissue macrophages, and dendritic cells (DCs), as well as their lineage-committed progenitors. 7 The primary function of mature MPS cells is the clearance of pathogens by phagocytosis. This activity is crucial during immune challenge to clear invasive pathogens. Mononuclear phagocytes also play an important role in the removal of apoptotic cell corpses, especially during embryonic development. In mice, embryonic macrop...

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The zebrafish spi1 promoter drives myeloid-specific expression in stable transgenic fish

Cited by 89 publications

References 23 publications

Single-cell transcriptome analysis of fish immune cells provides insight into the evolution of vertebrate immune cell types

Single-cell transcriptome analysis of fish immune cells provides insight into the evolution of vertebrate immune cell types

Leukocyte migration from a fish eye's view

Characterization of the mononuclear phagocyte system in zebrafish

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