Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Spring 5-15-2019 Defining Gastric Epithelial Cell Population Dynamics At Homeostasis And Following Injury Joseph Ronald Burclaff Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/art_sci_etds Part of the Biology Commons Recommended Citation Burclaff, Joseph Ronald, "Defining Gastric Epithelial Cell Population Dynamics At Homeostasis And Following Injury" (2019). Arts & Sciences Electronic Theses and Dissertations. 1778. https://openscholarship.wustl.edu/art_sci_etds/1778 This Dissertation is brought to you for free and open access by the Arts & Sciences at Washington University Open Scholarship. It has been accepted for inclusion in Arts & Sciences Electronic Theses and Dissertations by an authorized administrator of Washington University Open Scholarship. 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LOUIS Division of Biology and Biomedical Sciences Developmental, Regenerative, and Stem Cell Biology Dissertation Examination Committee: Jason Mills, Chair William Stenson, Co-Chair Matthew Ciorba Richard DiPaolo Gwendolyn Randolph Defining Gastric Epithelial Cell Population Dynamics At Homeostasis And Following Injury by Joseph Burclaff A dissertation presented to The Graduate School of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy May 2019 St. Louis, Missouri © 2019, Joseph Burclaff Table of Contents Acknowledgments........................................................................................................................ viii Abstract of the Dissertation ............................................................................................................ x Chapter 1: Introduction ................................................................................................................... 1 1.1 Introduction to plasticity ........................................................................................................... 2 1.2 Introduction to the stomach ....................................................................................................... 5 1.3 Gastric Plasticity ....................................................................................................................... 7 1.4 Gastric Tumorigensis ................................................................................................................ 9 1.5 Conclusion .............................................................................................................................. 10 Chapter 2: Targeted Parietal Cell Apoptosis Is Insufficient To Induce Metaplasia In The Stomach ....................................................................................................................................................... 13 2.1 Abstract ................................................................................................................................... 14 2.2 Parietal Cell Death and Metaplasia ......................................................................................... 14 2.3 Methods and Materials ............................................................................................................ 26 3.1 Abstract ......................................................................................................................................... 31 3.2 Introduction ................................................................................................................................... 32 3.3 Methods and Materials .................................................................................................................. 34 3.4 Chief cells take up little BrdU upon extended continuous pulsing ............................................... 36 3.5 BrdU administered throughout acute injury marks nearly all epithelial cells ............................... 39 3.6 Chief cells are not the predominate terminal maturation state for neck cells ............................... 43 3.7 Chief cells give rise to SPEM cells following gastric injury ........................................................ 45 3.8 Metaplastic cells can redifferentiate into chief cells following recovery from injury .................. 51 3.9 Discussion ..................................................................................................................................... 52 Chapter 4: Chief cells can differentiate in the absence of parietal cells ....................................... 61 4.1 Introduction ................................................................................................................................... 61 4.2 Results ........................................................................................................................................... 62 4.3 Discussion ..................................................................................................................................... 66 4.4 Methods and materials .................................................................................................................. 67 Chapter 5: MNU can be used to initiate gastric tumorigenesis .................................................... 70 5.1 Introduction ................................................................................................................................... 70 5.2 Results ........................................................................................................................................... 71 ii 5.3 Discussion ..................................................................................................................................... 76 5.4 Methods and Materials .................................................................................................................. 77 Chapter 6: Nitric Oxide may act as a damage signal between dying parietal cells and the gastric epithelium ..................................................................................................................................... 80 6.1 Introduction ................................................................................................................................... 80 6.2 Results ........................................................................................................................................... 82 6.3 Discussion ..................................................................................................................................... 87 6.4 Methods and Materials .................................................................................................................. 88 Chapter 7: Neck cells undergo many of the defined steps of Paligenosis when re-entering the cell cycle following injury ................................................................................................................... 92 7.1 Introduction ................................................................................................................................... 92 7.2 Results ........................................................................................................................................... 93 7.3 Discussion ..................................................................................................................................... 99 7.4 Methods and Materials ................................................................................................................ 100 Chapter 8: Conclusions and Future Directions ........................................................................... 103 8.1 Overall summary ......................................................................................................................... 103 8.2 Future directions ......................................................................................................................... 107 Appendix 1: Modeling murine gastric metaplasia through tamoxifen-induced acute parietal cell loss .............................................................................................................................................. 114 A1.1 Summary .................................................................................................................................. 115 A1.2 Introduction .............................................................................................................................. 115 A1.3 Materials ................................................................................................................................... 116 A1.4 Methods .................................................................................................................................... 117 A1.5 Notes ........................................................................................................................................ 121 A1.6 References ................................................................................................................................ 125 Appendix 2: Unintended targeting of Dmp1-Cre reveals a critical role for Bmpr1a signaling in the gastrointestinal mesenchyme of adult mice .......................................................................... 128 A2.1 Abstract ...................................................................................................................................