Isthmin 1 (ism1) is required for normal hematopoiesis in developing zebrafish.
Isthmin 1 (ism1) is required for normal hematopoiesis in developing zebrafish.
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DOI：
10.1371/journal.pone.0196872
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Stachura DL
中科院分区：
文献类型：
--
作者：
Berrun A;Harris E;Stachura DL
造血是从造血干细胞(HSCs)开始的一种基本的、受高度调控的生物学过程。在健康的生物体中，HSCs每天负责产生大量成熟的血细胞，但指示HSCs自我更新和分化为有丝分裂后血细胞的分子途径尚不完全清楚。为了了解这些分子途径，我们研究了斑马鱼(Danio Rerio)支持造血的细胞系中表达的新基因，这个模型系统越来越多地被用来揭示在其他脊椎动物物种发育中重要的分子途径。我们对来自胚胎和成年斑马鱼不同阶段的三个基质细胞系的转录组进行了RNA测序，并鉴定了数百个高表达的转录本。在我们的研究中，我们重点研究了isthmin1(Ism1)，因为它与人类基因同源基因具有共同的同源性，而且它是一种分泌性蛋白。为了确定ISM1的特征，我们进行了功能丧失实验，以确定成熟血细胞的产生是否受到干扰。用表达谱系特异性标记的转基因斑马鱼对髓系和红系谱系进行可视化和评分。Ism1基因敲除导致中性粒细胞、巨噬细胞和红细胞数量减少。来自ISM1变异体的克隆性甲基纤维素分析也显示，总的造血干细胞和祖细胞(HSPC)减少。总之，我们证明了ism1是斑马鱼造血过程中HSPC及其下游后代正常生成所必需的。进一步研究ISM1及其在造血中的重要性可能会阐明在进化中保守的造血过程，这些过程可以进一步研究其潜在的临床应用。
Hematopoiesis is an essential and highly regulated biological process that begins with hematopoietic stem cells (HSCs). In healthy organisms, HSCs are responsible for generating a multitude of mature blood cells every day, yet the molecular pathways that instruct HSCs to self-renew and differentiate into post-mitotic blood cells are not fully known. To understand these molecular pathways, we investigated novel genes expressed in hematopoietic-supportive cell lines from the zebrafish (Danio rerio), a model system increasingly utilized to uncover molecular pathways important in the development of other vertebrate species. We performed RNA sequencing of the transcriptome of three stromal cell lines derived from different stages of embryonic and adult zebrafish and identified hundreds of highly expressed transcripts. For our studies, we focused on isthmin 1 (ism1) due to its shared synteny with its human gene ortholog and because it is a secreted protein. To characterize ism1, we performed loss-of-function experiments to identify if mature blood cell production was disrupted. Myeloid and erythroid lineages were visualized and scored with transgenic zebrafish expressing lineage-specific markers. ism1 knockdown led to reduced numbers of neutrophils, macrophages, and erythrocytes. Analysis of clonal methylcellulose assays from ism1 morphants also showed a reduction in total hematopoietic stem and progenitor cells (HSPCs). Overall, we demonstrate that ism1 is required for normal generation of HSPCs and their downstream progeny during zebrafish hematopoiesis. Further investigation into ism1 and its importance in hematopoiesis may elucidate evolutionarily conserved processes in blood formation that can be further investigated for potential clinical utility.
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