Paralogs of Common Carp Granulocyte Colony-Stimulating Factor (G-CSF) Have Different Functions Regarding Development, Trafficking and Activation of Neutrophils
Paralogs of Common Carp Granulocyte Colony-Stimulating Factor (G-CSF) Have Different Functions Regarding Development, Trafficking and Activation of Neutrophils
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DOI：
10.3389/fimmu.2019.00255
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发表时间：
2019-02-19
影响因子：
7.3
通讯作者：
Moritomo, Tadaaki
中科院分区：
文献类型：
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作者：
Katakura, Fumihiko;Nishiya, Kohei;Moritomo, Tadaaki
哺乳动物粒细胞集落刺激因子（G-CSF; CSF 3）是一种促进中性粒细胞及其前体细胞发育、动员和活化的主要细胞因子。据报道，硬骨鱼具有两个旁系同源物，这可能是硬骨鱼全基因组复制（WGD）事件的结果，但对G-CSF旁系同源物的功能分歧仍然知之甚少。鲤鱼是一个异源四倍体物种，由于额外的WGD事件在鲤鱼谱系，在这里，我们报告的基因组同线性，序列相似性，和同源性的四个常见的鲤鱼G-CSF旁系（g-csfa 1和g-csfa 2; g-csfb 1和g-csfb 2）。g-csfa 1和g-csfa 2显示差异和相对较高的基因表达水平，而g-csfb 1和g-csfb 2在大多数组织中显示低基础基因表达水平。所有旁系同源物在巨噬细胞中的表达高于其他白细胞亚型，并且通过用有丝分裂原处理巨噬细胞而高度上调。重组G-CSFa 1和G-CSFb 1均能促进肾造血细胞增殖，但只有G-CSFb 1能诱导肾造血细胞向沿着细胞系分化。集落形成单位测定显示，G-CSFb 1单独刺激CFU-G集落形成的头和躯干肾，而G-CSFa 1和G-CSFb 1的组合刺激CFU-G和CFU-GM集落的形成。重组G-CSFa 1和G-CSFb 1也表现出对肾脏中性粒细胞的趋化活性，并且在G-CSFb 1刺激后中性粒细胞中cxcr 1 mRNA表达的上调最高。此外，G-CSFb 1比G-CSFa 1通过上调NADPH氧化酶组分p47 phox诱导肾中性粒细胞的启动。体内施用G-CSF旁系同源物增加鲤鱼循环血液中性粒细胞的数量。我们的研究结果表明，硬骨鱼的基因重复可以导致旁系同源物之间的功能分歧，并揭示了鲤科鱼类G-CSF旁系同源物的亚功能化。
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 p47phox. 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.