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Dynamic Analysis of Vascular Morphogenesis Using Transgenic Quail Embryos

Sato, Yuki and Poynter, Greg and Huss, David and Filla, Michael B. and Czirok, Andras and Rongish, Brenda J. and Little, Charles D. and Fraser, Scott E. and Lansford, Rusty (2010) Dynamic Analysis of Vascular Morphogenesis Using Transgenic Quail Embryos. PLoS ONE, 5 (9). Art. No. e12674 . ISSN 1932-6203. PMCID PMC2939056. https://resolver.caltech.edu/CaltechAUTHORS:20101004-091127769

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[img] Video (QuickTime) (Low magnification dynamic imaging of Tg(tie1:H2B-eYFPct2) quail embryo.) - Supplemental Material
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[img] Video (QuickTime) (Time-lapse movies showing Tg(tie1:H2B-eYFP) cell nuclei surrounded by QH1+ plasma membrane in EC cells. Tg(tie1:H2B-eYFP) quail embryos were injected with QH1-A647 at stage 7–8 and time-lapse captured every 13 minutes for 8.5 hours until 15 somites.) - Supplemental Material
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[img] Video (QuickTime) (Composite movie showing Tg(tie1:H2B-eYFP) cell nuclei (green) surrounded by QH1+ plasma membrane in EC cells.) - Supplemental Material
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[img] Video (QuickTime) (Imaris based cell tracking (xyt) of Tg(tie1:H2B-eYFP) time-lapse video. ECs arose at somite stage 7, 8, 9, 10, 11 are labeled by cyan, magenta, green, yellow, orange, respectively.) - Supplemental Material
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[img] Video (QuickTime) (Polygons are continuously rearranged. a) The area shown moves with the tissue, so the motion seen is autonomous.) - Supplemental Material
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[img] Video (QuickTime) (Time-lapse of Tg(tie1:H2B-eYFP) embryo shows anteriomedial rotation of newly assembled dorsal aortae. Embryos injected with QH1-A647 and imaged on the upright microscope with the dorsal side against the EC Agar culture using the 10× objective and 2×2 bi) - Supplemental Material
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[img] Video (QuickTime) (Cells move upstream (towards the heart, against the flow) and circulate within the walls of the dorsal aortae.) - Supplemental Material
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[img] Video (QuickTime) (Anteriomedial rotation of the dorsal aortae.) - Supplemental Material
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[img] Video (QuickTime) (2P microscopy of forming dorsal aortae from stage 11 through 12 (longitudinal view).) - Supplemental Material
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[img] Video (QuickTime) (Top. 2P microscopy of forming dorsal aortae from stage 11 through 12 (transverse view). ) - Supplemental Material
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Abstract

Background: One of the least understood and most central questions confronting biologists is how initially simple clusters or sheet-like cell collectives can assemble into highly complex three-dimensional functional tissues and organs. Due to the limits of oxygen diffusion, blood vessels are an essential and ubiquitous presence in all amniote tissues and organs. Vasculogenesis, the de novo self-assembly of endothelial cell (EC) precursors into endothelial tubes, is the first step in blood vessel formation [1]. Static imaging and in vitro models are wholly inadequate to capture many aspects of vascular pattern formation in vivo, because vasculogenesis involves dynamic changes of the endothelial cells and of the forming blood vessels, in an embryo that is changing size and shape. Methodology/Principal Findings: We have generated Tie1 transgenic quail lines Tg(tie1:H2B-eYFP) that express H2B-eYFP in all of their endothelial cells which permit investigations into early embryonic vascular morphogenesis with unprecedented clarity and insight. By combining the power of molecular genetics with the elegance of dynamic imaging, we follow the precise patterning of endothelial cells in space and time. We show that during vasculogenesis within the vascular plexus, ECs move independently to form the rudiments of blood vessels, all while collectively moving with gastrulating tissues that flow toward the embryo midline. The aortae are a composite of somatic derived ECs forming its dorsal regions and the splanchnic derived ECs forming its ventral region. The ECs in the dorsal regions of the forming aortae exhibit variable mediolateral motions as they move rostrally; those in more ventral regions show significant lateral-to-medial movement as they course rostrally. Conclusions/Significance: The present results offer a powerful approach to the major challenge of studying the relative role(s) of the mechanical, molecular, and cellular mechanisms of vascular development. In past studies, the advantages of the molecular genetic tools available in mouse were counterbalanced by the limited experimental accessibility needed for imaging and perturbation studies. Avian embryos provide the needed accessibility, but few genetic resources. The creation of transgenic quail with labeled endothelia builds upon the important roles that avian embryos have played in previous studies of vascular development.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1371/journal.pone.0012674 DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2939056/PubMed CentralArticle
ORCID:
AuthorORCID
Fraser, Scott E.0000-0002-5377-0223
Lansford, Rusty0000-0002-2159-3699
Additional Information:© 2010 Sato et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received December 19, 2009; Accepted June 30, 2010; Published September 14, 2010. Editor: Patrick Callaerts, Katholieke Universiteit Leuven, Belgium. Funding: Grants by the National Institutes of Health (NIH) National Center for Research Resources (R21HD047347-01), NIH National Heart, Lung, and Blood Institute (R01) HL085694 (BJR), HL068855 (CDL), HL87136 (AC), and NIH Center of Excellence in Genomic Science (P50 HG004071) supported the work. Y.S. was a postdoctoral fellow for research abroad of the Japan Society for the Promotion of Science (JSPS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank Le Trinh for critically reading the manuscript, John Choi for help with cell tracking and computational analysis, and members of the BIC for helpful discussions. Author Contributions: Conceived and designed the experiments: YS CDL RL. Performed the experiments: YS GP DH MBF RL. Analyzed the data: YS AC BJR CDL SEF RL. Contributed reagents/materials/analysis tools: AC. Wrote the paper: CDL SEF RL.
Funders:
Funding AgencyGrant Number
NIHR21HD047347-01
NIHR01 HL085694
NIHHL068855
NIHHL87136
NIHP50 HG004071
Subject Keywords:Developmental Biology
Issue or Number:9
PubMed Central ID:PMC2939056
Record Number:CaltechAUTHORS:20101004-091127769
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20101004-091127769
Official Citation:Sato Y, Poynter G, Huss D, Filla MB, Czirok A, et al. (2010) Dynamic Analysis of Vascular Morphogenesis Using Transgenic Quail Embryos. PLoS ONE 5(9): e12674. doi:10.1371/journal.pone.0012674
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:20268
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:17 Nov 2010 17:44
Last Modified:03 Oct 2019 02:07

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