Defining the ultrastructure of the hematopoietic stem cell niche by correlative light and electron microscopy
- University of Illinois at Chicago, United States
- University of California, San Diego, United States
- University of Wisconsin-Madison, United States
Abstract
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.
Data availability
SBEM datasets have been deposited in the National Center for Microscopy and Imaging Research (NCMIR) publicly accessible resource database Cell Image Library (CIL). There are six SBEM datasets (accession numbers: CIL:54845, CIL:54846, CIL:54847, CIL:54848, CIL:54849, CIL:54850) that are accessible as group with the following link: http://cellimagelibrary.org/groups/54850. CIL accession numbers are referenced in Table 1. Newly generated plasmids have been deposited in Addgene (#188944 and #188945).
Article and author information
Author details
Funding
National Heart, Lung, and Blood Institute (R01HL142998)
- Owen J Tamplin
National Institute of Diabetes and Digestive and Kidney Diseases (K01DK103908)
- Owen J Tamplin
American Heart Association (19POST34380221)
- Sobhika Agarwala
National Institute of Neurological Disorders and Stroke (1U24NS120055-01)
- Mark H Ellisman
National Institute of General Medical Sciences (R24 GM137200)
- Mark H Ellisman
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All experiments were performed in accordance with protocols approved by the Institutional Animal Care and Use Committees at the University of Illinois at Chicago (Protocol ACC 19-051) and the University of Wisconsin-Madison (Protocol M006348).
Copyright
© 2022, Agarwala et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Defining the ultrastructure of the hematopoietic stem cell niche by correlative light and electron microscopy
eLife 11:e64835.
https://doi.org/10.7554/eLife.64835
