Graduate Theses, Dissertations, and Problem Reports
2019
Canonical Wnt Mechanisms in Neural Crest Induction
Mark Stephen Perfetto West Virginia University, [email protected]
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Recommended Citation Perfetto, Mark Stephen, "Canonical Wnt Mechanisms in Neural Crest Induction" (2019). Graduate Theses, Dissertations, and Problem Reports. 7393. https://researchrepository.wvu.edu/etd/7393
This Dissertation is protected by copyright and/or related rights. It has been brought to you by the The Research Repository @ WVU with permission from the rights-holder(s). You are free to use this Dissertation in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you must obtain permission from the rights-holder(s) directly, unless additional rights are indicated by a Creative Commons license in the record and/ or on the work itself. This Dissertation has been accepted for inclusion in WVU Graduate Theses, Dissertations, and Problem Reports collection by an authorized administrator of The Research Repository @ WVU. For more information, please contact [email protected]. Canonical Wnt Mechanisms in Neural Crest Induction
Mark Perfetto
Dissertation submitted to the Eberly College of Arts and Sciences at West Virginia University
in partial fulfillment of the requirements for the degree of
Doctor of Philosophy in Biology
Shuo We, Ph.D., Chair Andrew Dacks, Ph.D Jennifer Gallagher, Ph.D. Peter Mathers, Ph.D. Eric Tucker, Ph.D.
Department of Biology
Morgantown, West Virginia December 2019
Keywords: Canonical Wnt Signaling, ADAMs, Akt signaling, neural crest
Copyright 2019 Mark Perfetto ABSTRACT
Canonical Wnt Mechanisms in Neural Crest Induction
Mark Perfetto
Canonical Wnt signaling is a pathway that is critical for normal development and the progression of disease. The canonical Wnt signaling pathway was put together carefully by synthesizing decades of research. Over those decades, canonical Wnt signaling was found to be crucial for nearly all aspects of development, but of importance to this thesis, is a key regulatory factor for the development of the highly migratory multipotent stem cells, the neural crest. As our knowledge of the importance of canonical Wnt signaling grew, research is being conducted to further our understanding of how canonical Wnt signaling communicates with other signaling pathways. Over my Ph.D., we have provided new mechanisms for how canonical Wnt signaling cross-talks with
Ephrin and Akt signaling. Here, I discuss how EphrinB signaling inhibits canonical Wnt signaling during neural crest induction and in human cell culture. Within that project, we found that the metalloproteinase ADAM19 protects ADAM13 during neural crest induction to inhibit EphrinB signaling, thus activating canonical Wnt signaling. I also discuss how Akt signaling is necessary for neural crest induction to activate canonical
Wnt signaling. Within that project we found that the RNA-helicase DDX3 is required for
Rac1 expression during neural crest induction to activate Akt signaling and thus activate canonical Wnt signaling. Finally, I discuss the impact of connecting canonical Wnt signaling with Ephrin and Akt signaling in neural crest, development, disease, and potential new discoveries and research. Table of contents:
Title page.………………………………………………………………………………………i
Thesis Abstract..……………………………………………………………………………….ii
Table of contents………………………………………………………………………………iii
Acknowledgements……………………………………………………………………………v
Chapter 1: Introduction………………………………………………………………………..1
Chapter 1.1: Canonical Wnt signaling………………………………………………1
Chapter 1.2: Akt signaling and canonical Wnt signaling crosstalk………………9
Chapter 1.3: The neural crest………………………………………………………..25
Chapter 1.4: ADAM metalloproteinases…………………………………………….34
Chapter 1.5: DEAD-box RNA helicases in development, canonical Wnt, and
Akt……………………………………………………………………………..……….39
Chapter 1.6: Figures………………………………………………………………….48
Chapter 2: Xenopus ADAM19 regulates Wnt signaling and neural crest specification by stabilizing ADAM13……………………………………………………………………………50
Abstract………………………………………………………………………………..51
Introduction……………………………………………………………………………52
Results…………………………………………………………………………………55
Discussion……………………………………………………………………………..67
Materials and methods……………………………………………………………….73
Acknowledgements…………………………………………………………………..77
Competing interests…………………………………………………………………..77
Tables…………………………………………………………………………………..78
iii Figures and legends………………………………………………………………….79
Chapter 3: The RNA helicase DDX3 induces neural crest through the Akt-GSK3β axis……………………………………………………………………………………………..95
Abstract……………………………………………………………………………….96
Introduction………………………………………………………………………….97
Results……………………………………………………………………………….101
Discussion…………………………………………………………………………..109
Materials and methods……………………………………………………………114
Acknowledgements………………………………………………………………..118
Competing interests………………………………………………………………118
Figures and legends……………………………………………………………….119
Chapter 4: Conclusions and future directions…………………………………………131
Literature cited………………………………………………………………………………142
iv Acknowledgements
This was hard. This was really hard. I absolutely love science and writing this dissertation challenged me in ways I had not expected and I have grown as a scientist and person because of it.
To my advisor Shuo Wei. You have taught me so much about science, but more importantly to me, you taught me how to be a scientist and have a life. You have been my north star and shown me infinite patience over our seven years together. Your tutelage has been invaluable, I will be forever grateful, and I hope to continue to learn from you and your example in the future.
To Jen Gallagher. You told me once that you thought that you could have been a better mentor to me. I am not sure how that is possible. You have put so much time