Doublecortin and CaM kinase-like-1 and leucine-rich-repeat-containing G-protein-coupled receptor mark quiescent and cycling intestinal stem cells, respectively.
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.
May
Randal
R
Department of Medicine,The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.
Sureban
Sripathi M
SM
Hoang
Nguyet
N
Riehl
Terrence E
TE
Lightfoot
Stan A
SA
Ramanujam
Rama
R
Wyche
James H
JH
Anant
Shrikant
S
Houchen
Courtney W
CW
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*
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
J Cell Biol. 1992 Oct;119(1):27-44
1527171
Curr Opin Cell Biol. 1994 Dec;6(6):795-803
7880525
Development. 1990 Dec;110(4):1001-20
2100251
Differentiation. 2003 Jan;71(1):28-41
12558601
Am J Anat. 1974 Dec;141(4):461-79
4440632
J Cell Sci. 1990 Sep;97 ( Pt 1):39-50
1701769
J Cell Biol. 1994 Sep;126(6):1547-64
8089185
Nature. 2007 Oct 25;449(7165):1003-7
17934449
J Biol Chem. 1992 Jul 25;267(21):15122-33
1634547
Gastroenterology. 2009 Aug;137(2):649-59, 659.e1-2
19445940
Histochem Cell Biol. 2008 Jun;129(6):705-33
18461349
Gastroenterology. 2008 May;134(5):1448-58
18471519
Gastroenterology. 2005 Nov;129(5):1567-80
16285956
Nat Struct Biol. 2003 May;10(5):324-33
12692530
Physiology (Bethesda). 2008 Dec;23:350-9
19074742
Proc Natl Acad Sci U S A. 2008 Mar 18;105(11):4358-63
18332421
J Theor Biol. 1987 Aug 21;127(4):381-91
3328018
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):3120-5
10077647
Cell. 1985 Feb;40(2):425-9
3967298
Bioessays. 2002 Jan;24(1):91-8
11782954
Nature. 2003 Oct 23;425(6960):836-41
14574412
Nat Rev Cancer. 2008 Jun;8(6):415-24
18480839
Am J Anat. 1981 Jan;160(1):93-103
7211719
Cell. 1990 Jun 29;61(7):1329-37
2364430
Nat Genet. 2008 Jul;40(7):915-20
18536716
Stem Cells. 2008 Mar;26(3):630-7
18055444
J Cell Biol. 1992 May;117(4):825-39
1349609
Eur J Cell Biol. 2003 May;82(5):262-70
12800981
Genomics. 1999 Feb 15;56(1):121-6
10036192
Am J Physiol Cell Physiol. 2009 Feb;296(2):C296-305
19073897
J Neurosci. 2000 Dec 15;20(24):9152-61
11124993
Int J Exp Pathol. 1997 Aug;78(4):219-43
9505935
Genes Dev. 2005 Oct 15;19(20):2412-7
16230531
Am J Anat. 1974 Dec;141(4):537-61
4440635
Proc Biol Sci. 1990 Jul 23;241(1300):13-8
1978330
Genes Dev. 2003 May 15;17(10):1253-70
12756227
Cancer Biol Ther. 2006 Apr;5(4):419-26
16575208
Int J Radiat Biol. 1988 Dec;54(6):1041-51
2903885
Science. 2001 Dec 7;294(5549):2155-8
11739954
J Cell Sci. 2002 Jun 1;115(Pt 11):2381-8
12006622
Development. 2003 Jun;130(12):2793-808
12736221
J Biol Chem. 2006 Apr 21;281(16):11292-300
16464855
ScientificWorldJournal. 2008 Nov 23;8:1168-76
19030762
Gastroenterology. 1999 Jan;116(1):7-14
9869596