PUBLICATION

Defining the ultrastructure of the hematopoietic stem cell niche by correlative light and electron microscopy

Authors
Agarwala, S., Kim, K.Y., Phan, S., Ju, S., Kong, Y.E., Castillon, G.A., Bushong, E.A., Ellisman, M.H., Tamplin, O.J.
ID
ZDB-PUB-220810-4
Date
2022
Source
eLIFE   11: (Journal)
Registered Authors
Tamplin, Owen
Keywords
developmental biology, regenerative medicine, stem cells, zebrafish
MeSH Terms
  • Zebrafish*
  • Animals
  • Microscopy, Electron
  • Hematopoietic Stem Cells/metabolism
  • Stem Cell Niche*
PubMed
35943143 Full text @ Elife
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.
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
cz2009TgTransgenic Insertion
    cz2010TgTransgenic Insertion
      la2TgTransgenic Insertion
        s896TgTransgenic Insertion
          zdf15TgTransgenic Insertion
            zf237TgTransgenic Insertion
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              Engineered Foreign Genes
              Marker Marker Type Name
              EGFPEFGEGFP
              mCherryEFGmCherry
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              Citation
              Agarwala, S., Kim, K.Y., Phan, S., Ju, S., Kong, Y.E., Castillon, G.A., Bushong, E.A., Ellisman, M.H., Tamplin, O.J. (2022) Defining the ultrastructure of the hematopoietic stem cell niche by correlative light and electron microscopy. eLIFE. 11:.