Exploring erythropoiesis of common carp (Cyprinus carpio) using an in vitro colony assay in the presence of recombinant carp kit ligand A and erythropoietin

Katakura, Fumihiko; Yabu, Takeshi; Yamaguchi, Takuya; Miyamae, Jiro; Shirinashihama, Yuki; Nakanishi, Teruyuki; Moritomo, Tadaaki

doi:10.1016/j.dci.2015.06.006

Cited by 9 publications

(9 citation statements)

References 51 publications

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“…Haematopoiesis in teleosts is similar to that in mammals, as demonstrated by several investigations in zebrafish ( Danio rerio )14, goldfish ( Carassius auratus L.)1516 and carp ( Cyprinus carpio )17. The zebrafish has recently emerged as a powerful genetic model of haematopoiesis1819.…”

mentioning

confidence: 82%

Mobilisation and dysfunction of haematopoietic stem/progenitor cells after Listonella anguillarum infection in ayu, Plecoglossus altivelis

Chen

Rong

et al. 2016

Sci Rep

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Haematopoietic stem/progenitor cells (HSPCs) can mobilise into blood and produce immune cell lineages following stress. However, the homeostasis and function of HSPCs after infection in teleosts are less well known. Here, we report that Listonella anguillarum infection enhances HSPC mobilisation and reduces their differentiation into myeloid cells in ayu (Plecoglossus altivelis), an aquacultured teleost in East Asia. We established a colony-forming unit culture (CFU-C) assay to measure HSPCs using conditioned medium from peripheral blood mononuclear cells stimulated with phytohaemagglutinin. The number of CFU-Cs decreased in the head kidney and increased in the blood and spleen of ayu infected with L. anguillarum. HSPC mobilisation after L. anguillarum infection was mediated by norepinephrine. Furthermore, HSPCs from ayu treated with L. anguillarum lipopolysaccharide (LPS) showed defective myeloid differentiation and could no longer rescue L. anguillarum-infected ayu. HSPC expansion was suppressed after L. anguillarum infection or its LPS treatment in vitro. These results reveal a link between HSPC regulation and pathogen infection in teleosts.

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mentioning

confidence: 82%

Mobilisation and dysfunction of haematopoietic stem/progenitor cells after Listonella anguillarum infection in ayu, Plecoglossus altivelis

Chen

Rong

et al. 2016

Sci Rep

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“…The most prominent cytokines that regulate proliferation and differentiation of erythroid cells in vertebrates are erythropoietin (EPO) [1][2][3] and Kit ligand (KITLG or stem cell factor, SCF) [4][5][6][7][8][9] . As was described in other vertebrates but not in zebrafish, binding of EPO and KITLG to their cognate receptors ensures erythroid lineage commitment by triggering specific signaling events 1,8,10,11 .…”

Section: Introductionmentioning

confidence: 82%

“…As a result, treatment of hematopoietic progenitor cells isolated from kidney marrow with Kitlga, but not Kitlgb, in combination with the other master regulator of erythroid lineage commitment, Epo 22,23 , enabled erythroid expansion of cells in suspension cultures and clonal assays, as in other vertebrates 1,8,10,11 . Next, we also tested the activity of dexamethasone (Dex) in addition to Epo and Kitlga.…”

Section: Discussionmentioning

confidence: 99%

Zebrafish Kit ligands cooperate with erythropoietin to promote erythroid cell expansion

Oltová

Svoboda

Svatonova

et al. 2020

Preprint

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Key points:• Kit signaling contributes to erythroid cell development and is conserved from fish to man • Ex vivo expansion and self-renewal of zebrafish erythroid progenitors requires addition of recombinant Kitlga AbstractKit ligand (Kitlg) is pleiotropic cytokine with a prominent role in vertebrate erythropoiesis. Although the role of Kitlg in this process has not yet been reported in Danio rerio (zebrafish), in the present study, we show that its function is evolutionary conserved. Zebrafish possess two copies of Kitlg genes (Kitlga and Kitlgb) due to whole genome duplication. To determine the role of each ligand in zebrafish, we performed a series of ex vivo and in vivo gain-and loss-of-function experiments. First, we tested the biological activity of recombinant Kitlg proteins in suspension culture from zebrafish whole kidney marrow and we demonstrate that Kitlga is necessary for expansion of erythroid progenitors ex vivo. To further address the role of kitlga and kitlgb in hematopoietic development in vivo, we performed gain-of-function experiments in zebrafish embryos, showing that both ligands cooperate with erythropoietin (Epo) to promote erythroid cell expansion. Finally, using the kita mutant (kita b5/b5 or sparse), we show that Kita receptor is crucial for Kitlga/b cooperation with Epo in erythroid cells. In summary, using optimized suspension culture conditions with recombinant cytokines (Epo, Kitlga), we are reporting for the first time ex vivo suspension cultures of zebrafish hematopoietic progenitor cells, which can serve as an indispensable tool to study normal and aberrant hematopoiesis in zebrafish. Furthermore, we conclude that although partial functional diversification of Kit ligands has been described in other processes, in erythroid development, both paralogs play a similar role and their function is evolutionary conserved.

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“…Recombinant carp erythropoietin (EPO), kit ligand A (KITLA) and thrombopoietin (TPO) were produced and purified as described previously (28, 29).…”

Section: Methodsmentioning

confidence: 99%

“…Freshly isolated leukocytes from carp kidney, spleen or peripheral blood were re-suspended to 2 × 10 5 cells/mL in E-RDF medium containing 40% FBS and 5% carp serum. Colony-forming unit assay using semi-solid media was carried out as previously described (28) with minor modifications. Briefly, a complete methylcellulose medium was prepared by mixing a 2.0% methylcellulose stock solution with an equal amount of the cell suspension.…”

Section: Methodsmentioning

confidence: 99%

Paralogs of Common Carp Granulocyte Colony-Stimulating Factor (G-CSF) Have Different Functions Regarding Development, Trafficking and Activation of Neutrophils

et al. 2019

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Mammalian granulocyte colony-stimulating factor (G-CSF; CSF3) is a primary cytokine that promotes the development, mobilization, and activation of neutrophils and their precursors. Teleosts have been reported to possess two paralogs as a likely result of the teleost-wide whole genome duplication (WGD) event, but functional divergence of G-CSF paralogs remains poorly understood. Common carp are an allotetraploid species owing to an additional WGD event in the carp lineage and here, we report on genomic synteny, sequence similarity, and phylogeny of four common carp G-CSF paralogs ( g-csfa1 and g-csfa2 ; g-csfb1 and g-csfb2 ). G-csfa1 and g-csfa2 show differential and relatively high gene expression levels, while g-csfb1 and g-csfb2 show low basal gene expression levels in most tissues. All paralogs are expressed higher in macrophages than in other leukocyte sub-types and are highly up-regulated by treatment of macrophages with mitogens. Recombinant G-CSFa1 and G-CSFb1 both promoted the proliferation of kidney hematopoietic cells, while only G-CSFb1 induced the differentiation of kidney cells along the neutrophil-lineage. Colony-forming unit assays revealed that G-CSFb1 alone stimulates the formation of CFU-G colonies from head- and trunk-kidney whereas the combination of G-CSFa1 and G-CSFb1 stimulates the formation of both CFU-G and CFU-GM colonies. Recombinant G-CSFa1 and G-CSFb1 also exhibit chemotactic activity against kidney neutrophils and up-regulation of cxcr1 mRNA expression was highest in neutrophils after G-CSFb1 stimulation. Furthermore, G-CSFb1 more than G-CSFa1 induced priming of kidney neutrophils through up-regulation of a NADPH-oxidase component p47 phox . In vivo administration of G-CSF paralogs increased the number of circulating blood neutrophils of carp. Our findings demonstrate that gene duplications in teleosts can lead to functional divergence between paralogs and shed light on the sub-functionalization of G-CSF paralogs in cyprinid fish.

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Exploring erythropoiesis of common carp (Cyprinus carpio) using an in vitro colony assay in the presence of recombinant carp kit ligand A and erythropoietin

Cited by 9 publications

References 51 publications

Mobilisation and dysfunction of haematopoietic stem/progenitor cells after Listonella anguillarum infection in ayu, Plecoglossus altivelis

Mobilisation and dysfunction of haematopoietic stem/progenitor cells after Listonella anguillarum infection in ayu, Plecoglossus altivelis

Zebrafish Kit ligands cooperate with erythropoietin to promote erythroid cell expansion

Paralogs of Common Carp Granulocyte Colony-Stimulating Factor (G-CSF) Have Different Functions Regarding Development, Trafficking and Activation of Neutrophils

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