TY - JOUR
T1 - Nestin-EGFP transgenic mice
T2 - Visualization of the self-renewal and multipotency of CNS stem cells
AU - Kawaguchi, Ayano
AU - Miyata, Takaki
AU - Sawamoto, Kazunobu
AU - Takashita, Noriko
AU - Murayama, Ayako
AU - Akamatsu, Wado
AU - Ogawa, Masaharu
AU - Okabe, Masaru
AU - Tano, Yasuo
AU - Goldman, Steven A.
AU - Okano, Hideyuki
N1 - Funding Information:
The authors thank Drs. Andrew McMahon for the Xh5 plasmid, Hiroyuki Sasaki for the Ass-hsp68-lacZ-pA plasmid, Miyuki Yamamoto for the RC2 antibody, Tatsunori Seki for the PSA-NCAM antibody, Hiromitsu Nakauchi and Yohei Morita for technical instructions for the cell sorting, and Anirvan Ghosh, Sean Morrison, Samuel Weiss, and Takuya Shimazaki for valuable comments on the manuscript. This work was supported by grants from the Ministry of Education, Science and Culture of Japan to H.O. and from the Human Frontier Science Program to H.O. and S.G., the Mathers Charitable Foundation, the National Multiple Sclerosis Society, and NINDS Grants R01NS29813 and R01NS33106 to S.G.
PY - 2001
Y1 - 2001
N2 - We generated transgenic mice carrying enhanced green fluorescent protein (EGFP) under the control of the nestin second-intronic enhancer (E/nestin:EGFP). Flow cytometry followed by in vitro assays revealed that in situ EGFP expression in the embryonic brain correlated with the mitotic index, the cogeneration of both neurons and gila, and the frequency of neurosphere formation in vitro. High-level EGFP expressors derived from embryos included a distinct subpopulation of cells that were self-renewable and multipotent, criteria that define neural stern cells (NSCs). Such cells were largely absent among lower-level or non-EGFP expressors, thereby permitting us to enrich for NSCs using EGFP expression level. In adults, although E/nestin:EGFP-positive cells included the NSC population, the frequency of neurosphere formation did not correlate directly with the level of EGFP expression. However, moderately EGFP-expressing cells in adults gained EGFP intensity when they formed neurospheres, suggesting embryonic and adult NSCs exist in different microenvironments in vivo.
AB - We generated transgenic mice carrying enhanced green fluorescent protein (EGFP) under the control of the nestin second-intronic enhancer (E/nestin:EGFP). Flow cytometry followed by in vitro assays revealed that in situ EGFP expression in the embryonic brain correlated with the mitotic index, the cogeneration of both neurons and gila, and the frequency of neurosphere formation in vitro. High-level EGFP expressors derived from embryos included a distinct subpopulation of cells that were self-renewable and multipotent, criteria that define neural stern cells (NSCs). Such cells were largely absent among lower-level or non-EGFP expressors, thereby permitting us to enrich for NSCs using EGFP expression level. In adults, although E/nestin:EGFP-positive cells included the NSC population, the frequency of neurosphere formation did not correlate directly with the level of EGFP expression. However, moderately EGFP-expressing cells in adults gained EGFP intensity when they formed neurospheres, suggesting embryonic and adult NSCs exist in different microenvironments in vivo.
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U2 - 10.1006/mcne.2000.0925
DO - 10.1006/mcne.2000.0925
M3 - Article
C2 - 11178865
AN - SCOPUS:0035118134
SN - 1044-7431
VL - 17
SP - 259
EP - 273
JO - Molecular and Cellular Neurosciences
JF - Molecular and Cellular Neurosciences
IS - 2
ER -
