TY - JOUR
T1 - Quantitative 3D imaging of the cranial microvascular environment at single-cell resolution
AU - Rindone, Alexandra N.
AU - Liu, Xiaonan
AU - Farhat, Stephanie
AU - Perdomo-Pantoja, Alexander
AU - Witham, Timothy F.
AU - Coutu, Daniel L.
AU - Wan, Mei
AU - Grayson, Warren L.
N1 - Funding Information:
We are appreciative of several individuals for their assistance with this study: Yuan Cai (light-sheet microscope training and alignment), Dr. Scot Kuo and Dr. Hoku West-Foyle (image software training and general advice), Jasmine Hu (staining protocol training), Shawna R. Synder (schematic illustration in Fig. 7a), and Dr. Jennifer Elisseeff (review of the manuscript). We also thank the JHU Integrated Imaging Center and JHU SOM Microscope Facility for providing the equipment and software necessary to carry out this work. This work was supported by the National Science Foundation Graduate Research Fellowship (A.N.R.), NIH National Institute for Dental and Craniofacial Research Grant No. 5 F31 DE029109-02 (A.N.R.) and 5 R01 DE027957-02 (W.L.G.), ARCS Foundation Metropolitan Washington Chapter (A.N.R.), and NIH Shared Instrumentation Grant No. 1S10OD020152-01A1 (Integrated Imaging Center).
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - Vascularization is critical for skull development, maintenance, and healing. Yet, there remains a significant knowledge gap in the relationship of blood vessels to cranial skeletal progenitors during these processes. Here, we introduce a quantitative 3D imaging platform to enable the visualization and analysis of high-resolution data sets (>100 GB) throughout the entire murine calvarium. Using this technique, we provide single-cell resolution 3D maps of vessel phenotypes and skeletal progenitors in the frontoparietal cranial bones. Through these high-resolution data sets, we demonstrate that CD31hiEmcnhi vessels are spatially correlated with both Osterix+ and Gli1+ skeletal progenitors during postnatal growth, healing, and stimulated remodeling, and are concentrated at transcortical canals and osteogenic fronts. Interestingly, we find that this relationship is weakened in mice with a conditional knockout of PDGF-BB in TRAP+ osteoclasts, suggesting a potential role for osteoclasts in maintaining the native cranial microvascular environment. Our findings provide a foundational framework for understanding how blood vessels and skeletal progenitors spatially interact in cranial bone, and will enable more targeted studies into the mechanisms of skull disease pathologies and treatments. Additionally, our technique can be readily adapted to study numerous cell types and investigate other elusive phenomena in cranial bone biology.
AB - Vascularization is critical for skull development, maintenance, and healing. Yet, there remains a significant knowledge gap in the relationship of blood vessels to cranial skeletal progenitors during these processes. Here, we introduce a quantitative 3D imaging platform to enable the visualization and analysis of high-resolution data sets (>100 GB) throughout the entire murine calvarium. Using this technique, we provide single-cell resolution 3D maps of vessel phenotypes and skeletal progenitors in the frontoparietal cranial bones. Through these high-resolution data sets, we demonstrate that CD31hiEmcnhi vessels are spatially correlated with both Osterix+ and Gli1+ skeletal progenitors during postnatal growth, healing, and stimulated remodeling, and are concentrated at transcortical canals and osteogenic fronts. Interestingly, we find that this relationship is weakened in mice with a conditional knockout of PDGF-BB in TRAP+ osteoclasts, suggesting a potential role for osteoclasts in maintaining the native cranial microvascular environment. Our findings provide a foundational framework for understanding how blood vessels and skeletal progenitors spatially interact in cranial bone, and will enable more targeted studies into the mechanisms of skull disease pathologies and treatments. Additionally, our technique can be readily adapted to study numerous cell types and investigate other elusive phenomena in cranial bone biology.
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U2 - 10.1038/s41467-021-26455-w
DO - 10.1038/s41467-021-26455-w
M3 - Article
C2 - 34711819
AN - SCOPUS:85118453466
SN - 2041-1723
VL - 12
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 6219
ER -