Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations
Continual renewal of the intestinal epithelium is dependent on active- and slow-cycling stem cells that are confined to the crypt base. Tight regulation of these stem cell populations maintains homeostasis by balancing proliferation and differentiation to support critical intestinal functions. The h...
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doaj-fddeaf7b1f954bfe884f0a06244f9c052020-11-25T00:11:54ZengElsevierCellular and Molecular Gastroenterology and Hepatology2352-345X2018-01-0161799610.1016/j.jcmgh.2018.02.007Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell PopulationsNicholas R. Smith0John R. Swain1Paige S. Davies2Alexandra C. Gallagher3Michael S. Parappilly4Catherine Z. Beach5Philip R. Streeter6Ian A. Williamson7Scott T. Magness8Melissa H. Wong9Department of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonDepartment of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonDepartment of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonDepartment of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonDepartment of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonDepartment of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonDepartment of Pediatrics, Oregon Health & Science University, Portland, OregonDepartment of Biomedical Engineering, Department of Medicine, Department of Cell Biology and Physiology, University of North Carolina, Chapel Hill, North CarolinaDepartment of Biomedical Engineering, Department of Medicine, Department of Cell Biology and Physiology, University of North Carolina, Chapel Hill, North CarolinaDepartment of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, OregonContinual renewal of the intestinal epithelium is dependent on active- and slow-cycling stem cells that are confined to the crypt base. Tight regulation of these stem cell populations maintains homeostasis by balancing proliferation and differentiation to support critical intestinal functions. The hierarchical relation of discrete stem cell populations in homeostasis or during regenerative epithelial repair remains controversial. Although recent studies have supported a model for the active-cycling leucine-rich repeat-containing G-protein–coupled receptor 5 (Lgr5)+ intestinal stem cell (ISC) functioning upstream of the slow-cycling B lymphoma Mo-MLV insertion region 1 homolog (Bmi1)-expressing cell, other studies have reported the opposite relation. Tools that facilitate simultaneous analyses of these populations are required to evaluate their coordinated function. Methods: We used novel monoclonal antibodies (mAbs) raised against murine intestinal epithelial cells in conjunction with ISC–green fluorescent protein (GFP) reporter mice to analyze relations between ISC populations by microscopy. Ex vivo 3-dimensional cultures, flow cytometry, and quantitative reverse-transcription polymerase chain reaction analyses were performed. Results: Two novel mAbs recognized distinct subpopulations of the intestinal epithelium and when used in combination permitted isolation of discrete Lgr5GFP and Bmi1GFP-enriched populations with stem activity. Growth from singly isolated Lgr5GFP ISCs gave rise to small spheroids. Spheroids did not express Lgr5GFP and instead up-regulated Bmi1GFP expression. Conversely, Bmi1-derived spheroids initiated Lgr5GFP expression as crypt domains were established. Conclusions: These data showed the functional utility of murine mAbs in the isolation and investigation of Lgr5GFP and Bmi1GFP ISC-enriched populations. Ex vivo analyses showed hierarchical plasticity between different ISC-expressing states; specifically Lgr5GFP ISCs gave rise to Bmi1GFP cells, and vice versa. These data highlight the impact of temporal and physiological context on unappreciated interactions between Lgr5GFP and Bmi1GFP cells during crypt formation.http://www.sciencedirect.com/science/article/pii/S2352345X18300444Intestinal Stem CellsHierarchyLgr5Bmi1Plasticity |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nicholas R. Smith John R. Swain Paige S. Davies Alexandra C. Gallagher Michael S. Parappilly Catherine Z. Beach Philip R. Streeter Ian A. Williamson Scott T. Magness Melissa H. Wong |
spellingShingle |
Nicholas R. Smith John R. Swain Paige S. Davies Alexandra C. Gallagher Michael S. Parappilly Catherine Z. Beach Philip R. Streeter Ian A. Williamson Scott T. Magness Melissa H. Wong Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations Cellular and Molecular Gastroenterology and Hepatology Intestinal Stem Cells Hierarchy Lgr5 Bmi1 Plasticity |
author_facet |
Nicholas R. Smith John R. Swain Paige S. Davies Alexandra C. Gallagher Michael S. Parappilly Catherine Z. Beach Philip R. Streeter Ian A. Williamson Scott T. Magness Melissa H. Wong |
author_sort |
Nicholas R. Smith |
title |
Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations |
title_short |
Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations |
title_full |
Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations |
title_fullStr |
Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations |
title_full_unstemmed |
Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations |
title_sort |
monoclonal antibodies reveal dynamic plasticity between lgr5- and bmi1-expressing intestinal cell populations |
publisher |
Elsevier |
series |
Cellular and Molecular Gastroenterology and Hepatology |
issn |
2352-345X |
publishDate |
2018-01-01 |
description |
Continual renewal of the intestinal epithelium is dependent on active- and slow-cycling stem cells that are confined to the crypt base. Tight regulation of these stem cell populations maintains homeostasis by balancing proliferation and differentiation to support critical intestinal functions. The hierarchical relation of discrete stem cell populations in homeostasis or during regenerative epithelial repair remains controversial. Although recent studies have supported a model for the active-cycling leucine-rich repeat-containing G-protein–coupled receptor 5 (Lgr5)+ intestinal stem cell (ISC) functioning upstream of the slow-cycling B lymphoma Mo-MLV insertion region 1 homolog (Bmi1)-expressing cell, other studies have reported the opposite relation. Tools that facilitate simultaneous analyses of these populations are required to evaluate their coordinated function.
Methods: We used novel monoclonal antibodies (mAbs) raised against murine intestinal epithelial cells in conjunction with ISC–green fluorescent protein (GFP) reporter mice to analyze relations between ISC populations by microscopy. Ex vivo 3-dimensional cultures, flow cytometry, and quantitative reverse-transcription polymerase chain reaction analyses were performed.
Results: Two novel mAbs recognized distinct subpopulations of the intestinal epithelium and when used in combination permitted isolation of discrete Lgr5GFP and Bmi1GFP-enriched populations with stem activity. Growth from singly isolated Lgr5GFP ISCs gave rise to small spheroids. Spheroids did not express Lgr5GFP and instead up-regulated Bmi1GFP expression. Conversely, Bmi1-derived spheroids initiated Lgr5GFP expression as crypt domains were established.
Conclusions: These data showed the functional utility of murine mAbs in the isolation and investigation of Lgr5GFP and Bmi1GFP ISC-enriched populations. Ex vivo analyses showed hierarchical plasticity between different ISC-expressing states; specifically Lgr5GFP ISCs gave rise to Bmi1GFP cells, and vice versa. These data highlight the impact of temporal and physiological context on unappreciated interactions between Lgr5GFP and Bmi1GFP cells during crypt formation. |
topic |
Intestinal Stem Cells Hierarchy Lgr5 Bmi1 Plasticity |
url |
http://www.sciencedirect.com/science/article/pii/S2352345X18300444 |
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