Prospective identification of functionally distinct stem cells and neurosphere-initiating cells in adult mouse forebrain.
Neurosphere formation is commonly used as a surrogate for neural stem cell (NSC) function but the relationship between neurosphere-initiating cells (NICs) and NSCs remains unclear. We prospectively identified, and isolated by flow cytometry, adult mouse lateral ventricle subventricular zone (SVZ) NICs as Glast(mid)EGFR(high)PlexinB2(high)CD24(-/low)O4/PSA-NCAM(-/low)Ter119/CD45(-) (GEPCOT) cells. They were highly mitotic and short-lived in vivo based on fate-mapping with Ascl1(CreERT2) and Dlx1(CreERT2). In contrast, pre-GEPCOT cells were quiescent, expressed higher Glast, and lower EGFR and PlexinB2. Pre-GEPCOT cells could not form neurospheres but expressed the stem cell markers Slc1a3-CreER(T), GFAP-CreER(T2), Sox2(CreERT2), and Gli1(CreERT2) and were long-lived in vivo. While GEPCOT NICs were ablated by temozolomide, pre-GEPCOT cells survived and repopulated the SVZ. Conditional deletion of the Bmi-1 polycomb protein depleted pre-GEPCOT and GEPCOT cells, though pre-GEPCOT cells were more dependent upon Bmi-1 for Cdkn2a (p16(Ink4a)) repression. Our data distinguish quiescent NSCs from NICs and make it possible to study their properties in vivo.DOI: http://dx.doi.org/10.7554/eLife.02669.001.
Department of Pediatrics, Children's Research Institute, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, United States.
John K
JK
Mich
Department of Pediatrics, Children's Research Institute, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, United States.
Robert Aj
RA
Signer
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States.
Daisuke
D
Nakada
Department of Pediatrics, Children's Research Institute, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, United States.
André
A
Pineda
Department of Pediatrics, Children's Research Institute, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, United States.
Rebecca J
RJ
Burgess
Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, United States.
Tou Yia
TY
Vue
Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, United States.
Jane E
JE
Johnson
Department of Pediatrics, Children's Research Institute, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, United States sean.morrison@utsouthwestern.edu.
Sean J
SJ
Morrison
D000375:Aging / Q000378:metabolism*; D000818:Animals; D050256:Antimitotic Agents / Q000494:pharmacology; D049109:Cell Proliferation; D002469:Cell Separation; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D019426:Integrases / Q000378:metabolism; D008810:Mice, Inbred C57BL; D058953:Neural Stem Cells / Q000166:cytology*; D055495:Neurogenesis / Q000187:drug effects; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D063150:Polycomb Repressive Complex 1 / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D018874:Spheroids, Cellular / Q000166:cytology*; D000077204:Temozolomide
D050256:Antimitotic Agents; C489421:Bmi1 protein, mouse; D005904:Glial Fibrillary Acidic Protein; D011518:Proto-Oncogene Proteins; D003606:Dacarbazine; D063150:Polycomb Repressive Complex 1; C045073:Cre recombinase; D019426:Integrases; D000077204:Temozolomide
Bmi-1; fate-mapping; forebrain; prospective identification; stem cell