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Retroviral Lineage Analysis of the Vagal Neural Crest Reveals Multipotency Towards the Cardiac and Enteric Fates

Citation

Tang, Weiyi (2022) Retroviral Lineage Analysis of the Vagal Neural Crest Reveals Multipotency Towards the Cardiac and Enteric Fates. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/qakz-vm04. https://resolver.caltech.edu/CaltechTHESIS:04162022-233242577

Abstract

The neural crest is a migratory stem cell population that gives rise to the craniofacial skeleton, heart septa, pigment cells, and peripheral nervous system. Defects in neural crest development can lead to a broad range of congenital diseases, e.g., persistent truncus arteriosus, characterized by a mixture of oxygenated and deoxygenated blood, is related to the absence of the neural crest-derived outflow tract septum. Thus, a thorough understanding about neural crest migration, differentiation, and cell fate can shine lights on diagnosis and treatment of many congenital defects. A long-standing question is whether neural crest cells are composed of multipotent cells capable of giving rise to a wide range of cell types, or a mixture of fate-determined cells migrating to their destinations. Avian embryos resemble humans during neural crest development, but are more accessible to experimental manipulations than mammalian models, making them an ideal model to study the neural crest. Despite the abundance of information obtained from elegant experiments through interspecies grafting, the avian model lacks a direct tool to determine whether these cells are multipotent in vivo.

Here, we present a new clonal analysis tool that takes advantage of Replication Incompetent Avian retroviruses (RIAs). We validate the method in vitro and present the potential application in the chick embryo to test the multipotency of the trunk neural crest. Next, we perform RIA-mediated lineage tracing at a population level and uncover cardiomyocytes as a previously unknown cardiac neural crest derivative in both chicken and mouse. Furthermore, we utilize RIA-mediated clonal analysis to identify individual premigratory vagal neural crest cells as a multipotent stem cell that forms cell types in both the heart and the gut. We then confirm the results by single-cell photoconversion assay that further confirms that migrating neural crest cells are also multipotent. Time-lapse imaging shows that stochastic post-mitotic migration is a cellular mechanism underlying multipotency. Finally, molecular perturbation experiments show that CXCR4 and RET are essential guidance cues for migratory neural crest cells to enter the heart and the gut, respectively. Together, these results demonstrate the utility of using RIA viruses to tackle questions regarding the lineage, developmental potential, and migratory pathways followed by neural crest cells in avian embryos.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:Developmental biology, neural crest, clonal analysis, multipotency, cardiac, enteric nervous system, RIAs
Degree Grantor:California Institute of Technology
Division:Biology and Biological Engineering
Major Option:Biology
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Bronner, Marianne E.
Thesis Committee:
  • Stathopoulos, Angelike (chair)
  • Rothenberg, Ellen V.
  • Thomson, Matthew
  • Bronner, Marianne E.
Defense Date:4 March 2022
Record Number:CaltechTHESIS:04162022-233242577
Persistent URL:https://resolver.caltech.edu/CaltechTHESIS:04162022-233242577
DOI:10.7907/qakz-vm04
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.22002/D1.20102DOIMovie 2-1
http://dx.doi.org/10.22002/D1.20103DOISupplementary data 4-1
http://dx.doi.org/10.22002/D1.20104DOISupplementary data 4-2
http://dx.doi.org/10.22002/D1.20105DOISupplementary Movie 4-1
http://dx.doi.org/10.22002/D1.20106DOISupplementary Movie 4-2
http://dx.doi.org/10.22002/D1.20107DOISupplementary Movie 4-3
http://dx.doi.org/10.22002/D1.20108DOISupplementary Movie 4-4
http://dx.doi.org/10.1242/dev.193193DOITang, W. & Bronner, M. E. Neural crest lineage analysis: From past to future trajectory. Development 147, (2020)
http://dx.doi.org/10.1016/j.ydbio.2019.03.007DOITang, W., Li, Y., Gandhi, S. & Bronner, M. E. Multiplex clonal analysis in the chick embryo using retrovirally-mediated combinatorial labeling. Dev. Biol. 450, (2019)
http://dx.doi.org/10.7554/eLife.47929DOITang, W.*, Martik, M. L.*, Li, Y. & Bronner, M. E. Cardiac neural crest contributes to cardiomyocytes in amniotes and heart regeneration in zebrafish. Elife 8, (2019)
http://dx.doi.org/10.1038/s41467-021-22146-8DOITang, W., Li, Y., Li, A. & Bronner, M. E. Clonal analysis and dynamic imaging identify multipotency of individual Gallus gallus caudal hindbrain neural crest cells toward cardiac and enteric fates. Nat. Commun. 12, (2021)
ORCID:
AuthorORCID
Tang, Weiyi0000-0002-1279-1001
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:14551
Collection:CaltechTHESIS
Deposited By: Weiyi Tang
Deposited On:20 Apr 2022 19:42
Last Modified:27 Apr 2022 19:14

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