title: Doublecortin and CaM kinase-like-1 and leucine-rich-repeat-containing G-protein-coupled receptor mark quiescent and cycling intestinal stem cells, respectively.
abstract: It is thought that small intestinal epithelia (IE) undergo continuous self-renewal primarily due to their population of undifferentiated stem cells. These stem cells give rise to transit amplifying (daughter/progenitor) cells, which can differentiate into all mature cell types required for normal gut function. Identification of stem cells in IE is paramount to fully understanding this renewal process. One major obstacle in gastrointestinal stem cell biology has been the lack of definitive markers that identify small intestinal stem cells (ISCs). Here we demonstrate that the novel putative ISC marker doublecortin and CaM kinase-like-1 (DCAMKL-1) is predominantly expressed in quiescent cells in the lower two-thirds of intestinal crypt epithelium and in occasional crypt-based columnar cells (CBCs). In contrast, the novel putative stem cell marker leucine-rich-repeat-containing G-protein-coupled receptor (LGR5) is observed in rapidly cycling CBCs and in occasional crypt epithelial cells. Furthermore, functionally quiescent DCAMKL-1+ crypt epithelial cells retain bromo-deoxyuridine in a modified label retention assay. Moreover, we demonstrate that DCAMKL-1 is a cell surface expressing protein; DCAMKL-1+ cells, isolated from the adult mouse small intestine by fluorescence activated cell sorting, self-renew and ultimately form spheroids in suspension culture. These spheroids formed glandular epithelial structures in the flanks of athymic nude mice, which expressed multiple markers of gut epithelial lineage. Thus, DCAMKL-1 is a marker of quiescent ISCs and can be distinguished from the cycling stem/progenitors (LGR5+). Moreover, DCAMKL-1 can be used to isolate normal small intestinal stem cells and represents a novel research tool for regenerative medicine and cancer therapy.
authors.affiliation: Department of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.
authors.forename: Randal
authors.initials: R
authors.lastname: May
authors.forename: Sripathi M
authors.initials: SM
authors.lastname: Sureban
authors.forename: Nguyet
authors.initials: N
authors.lastname: Hoang
authors.forename: Terrence E
authors.initials: TE
authors.lastname: Riehl
authors.forename: Stan A
authors.initials: SA
authors.lastname: Lightfoot
authors.forename: Rama
authors.initials: R
authors.lastname: Ramanujam
authors.forename: James H
authors.initials: JH
authors.lastname: Wyche
authors.forename: Shrikant
authors.initials: S
authors.lastname: Anant
authors.forename: Courtney W
authors.initials: CW
authors.lastname: Houchen
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000032:analysis / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D001973:Bromodeoxyuridine; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D045744:Cell Line, Tumor; D019070:Cell Lineage / Q000502:physiology; D049109:Cell Proliferation; D000089742:Doublecortin-Like Kinases; D005434:Flow Cytometry; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000032:analysis / Q000378:metabolism*; D018874:Spheroids, Cellular; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: D000954:Antigens, Surface; D015415:Biomarkers; C493597:Lgr5 protein, mouse; D043562:Receptors, G-Protein-Coupled; D000089742:Doublecortin-Like Kinases; C485525:Dclk1 protein, mouse; D017346:Protein Serine-Threonine Kinases; D001973:Bromodeoxyuridine
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