Defining the ultrastructure of the hematopoietic stem cell niche by correlative light and electron microscopy.
Defining the ultrastructure of the hematopoietic stem cell niche by correlative light and electron microscopy.
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DOI：
10.7554/elife.64835
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发表时间：
2022-08-09
期刊：
影响因子：
7.7
通讯作者：
Tamplin, Owen J.
中科院分区：
文献类型：
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作者：
Agarwala, Sobhika;Kim, Keun-Young;Phan, Sebastien;Ju, Saeyeon;Kong, Ye Eun;Castillon, Guillaume A.;Bushong, Eric A.;Ellisman, Mark H.;Tamplin, Owen J.
血液系统由造血干细胞和祖细胞（HSPCs）支持，这些造血干细胞和祖细胞存在于一个被称为生态位的特殊微环境中。许多不同的生态位细胞类型支持HSPCs，但它们如何相互作用及其超微结构一直难以确定。在这里，我们发现单个内源性HSPCs可以通过光学显微镜跟踪，然后通过连续块面扫描电子显微镜（SBEM）在多尺度水平上进行识别。以斑马鱼幼鱼肾骨髓（KM）生态位为模型，我们通过薄层显微镜跟踪单荧光标记的HSPCs，然后在3D SBEM数据集中确认它们的确切位置。我们发现了多种不同形态的HSPC及其周围的生态位细胞，这表明HSPC沉积位点可能存在功能异质性。我们的方法还允许我们鉴定多巴胺β -羟化酶（dbh）阳性神经节细胞是HSPC生态位中以前未表征的功能细胞类型。通过整合多种成像方式，我们可以解析活组织深处单个稀有细胞的超微结构，并确定HSPC与其周围生态位细胞类型之间的所有接触。
The blood system is supported by hematopoietic stem and progenitor cells (HSPCs) found in a specialized microenvironment called the niche. Many different niche cell types support HSPCs, however how they interact and their ultrastructure has been difficult to define. Here, we show that single endogenous HSPCs can be tracked by light microscopy, then identified by serial block-face scanning electron microscopy (SBEM) at multiscale levels. Using the zebrafish larval kidney marrow (KM) niche as a model, we followed single fluorescently labeled HSPCs by light sheet microscopy, then confirmed their exact location in a 3D SBEM dataset. We found a variety of different configurations of HSPCs and surrounding niche cells, suggesting there could be functional heterogeneity in sites of HSPC lodgement. Our approach also allowed us to identify dopamine beta-hydroxylase (dbh) positive ganglion cells as a previously uncharacterized functional cell type in the HSPC niche. By integrating multiple imaging modalities, we could resolve the ultrastructure of single rare cells deep in live tissue and define all contacts between an HSPC and its surrounding niche cell types.