title: Single-cell resolution of morphological changes in hemogenic endothelium.
abstract: Endothelial-to-hematopoietic transition (EHT) occurs within a population of hemogenic endothelial cells during embryogenesis, and leads to the formation of the adult hematopoietic system. Currently, the prospective identification of specific endothelial cells that will undergo EHT, and the cellular events enabling this transition, are not known. We set out to define precisely the morphological events of EHT, and to correlate cellular morphology with the expression of the transcription factors RUNX1 and SOX17. A novel strategy was developed to allow for correlation of immunofluorescence data with the ultrastructural resolution of scanning electron microscopy. The approach can identify single endothelial cells undergoing EHT, as identified by the ratio of RUNX1 to SOX17 immunofluorescence levels, and the morphological changes associated with the transition. Furthermore, this work details a new technical resource that is widely applicable for correlative analyses of single cells in their native tissue environments.
authors.affiliation: Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: Frank L
authors.initials: FL
authors.lastname: Bos
authors.affiliation: Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: John S
authors.initials: JS
authors.lastname: Hawkins
authors.affiliation: Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA 94158, USA Department of Pediatrics, Division of Neonatology, University of California San Francisco School of Medicine, San Francisco, CA 94143, USA ann.zovein@ucsf.edu.
authors.forename: Ann C
authors.initials: AC
authors.lastname: Zovein
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D055018:Hemangioblasts / Q000166:cytology*; D006413:Hematopoietic System / Q000196:embryology* / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D055760:SOXF Transcription Factors / Q000378:metabolism*; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D050676:Core Binding Factor Alpha 2 Subunit; C493728:RUNX1 protein, human; D055760:SOXF Transcription Factors
keywords: Endothelial-to-hematopoietic transition; Endothelium; Hematopoiesis; Hemogenic; Human; Imaging; Mice; RUNX1; SOX17; Scanning electron microscopy