TY - JOUR
T1 - Stem Cell Defects in ATM-Deficient Undifferentiated Spermatogonia through DNA Damage-Induced Cell-Cycle Arrest
AU - Takubo, Keiyo
AU - Ohmura, Masako
AU - Azuma, Masaki
AU - Nagamatsu, Go
AU - Yamada, Wakako
AU - Arai, Fumio
AU - Hirao, Atsushi
AU - Suda, Toshio
N1 - Funding Information:
We thank Drs. P. J. McKinnon, P. Leder, and M. Okabe for transgenic mice. We also thank Drs. H. Saya, T. Kitamura, and A. Iwama for fruitful discussions, T. Ogawa for critical reading, and Ms. A. Kumakubo, Ms. A. Ono, and Ms. T. Hirose for essential support. K.T. is a research fellow of the Japan Society for the Promotion of Science. A.H. was supported by grants-in-aid from the Stem Cell Research of the Ministry of Education, Science, Sports, and Culture, Japan, and a CREST grant by the JST. T.S. was supported by a grant-in-aid from the Specially Promoted Research of the Ministry of Education, Science, Sports, and Culture, Japan.
PY - 2008/2/7
Y1 - 2008/2/7
N2 - Mammalian spermatogenesis is maintained by stem cell capacity within undifferentiated spermatogonial subpopulation. Here, using a combination of surface markers, we describe a purification method for undifferentiated spermatogonia. Flow cytometric analysis revealed that this population is composed of Plzf-positive cells and exhibits quiescence and the side population phenotype, fulfilling general stem cell criteria. We then applied this method to analyze undifferentiated spermatogonia and stem cell activity of Atm-/- mice. Atm-/- testis shows progressive depletion of undifferentiated spermatogonia accompanied by cell-cycle arrest. In Atm-/- undifferentiated spermatogonia, a self-renewal defect was observed in vitro and in vivo. Accumulation of DNA damage and activation of the p19Arf-p53-p21Cip1/Waf1 pathway were observed in Atm-/- undifferentiated spermatogonia. Moreover, suppression of p21Cip1/Waf1 in an Atm-/- background restored transplantation ability of undifferentiated spermatogonia, indicating that ATM plays an essential role in maintenance of undifferentiated spermatogonia and their stem cell capacity by suppressing DNA damage-induced cell-cycle arrest.
AB - Mammalian spermatogenesis is maintained by stem cell capacity within undifferentiated spermatogonial subpopulation. Here, using a combination of surface markers, we describe a purification method for undifferentiated spermatogonia. Flow cytometric analysis revealed that this population is composed of Plzf-positive cells and exhibits quiescence and the side population phenotype, fulfilling general stem cell criteria. We then applied this method to analyze undifferentiated spermatogonia and stem cell activity of Atm-/- mice. Atm-/- testis shows progressive depletion of undifferentiated spermatogonia accompanied by cell-cycle arrest. In Atm-/- undifferentiated spermatogonia, a self-renewal defect was observed in vitro and in vivo. Accumulation of DNA damage and activation of the p19Arf-p53-p21Cip1/Waf1 pathway were observed in Atm-/- undifferentiated spermatogonia. Moreover, suppression of p21Cip1/Waf1 in an Atm-/- background restored transplantation ability of undifferentiated spermatogonia, indicating that ATM plays an essential role in maintenance of undifferentiated spermatogonia and their stem cell capacity by suppressing DNA damage-induced cell-cycle arrest.
KW - STEMCELL
UR - http://www.scopus.com/inward/record.url?scp=38649099103&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=38649099103&partnerID=8YFLogxK
U2 - 10.1016/j.stem.2007.10.023
DO - 10.1016/j.stem.2007.10.023
M3 - Article
C2 - 18371438
AN - SCOPUS:38649099103
VL - 2
SP - 170
EP - 182
JO - Cell Stem Cell
JF - Cell Stem Cell
SN - 1934-5909
IS - 2
ER -