TY - JOUR
T1 - Differentiation of glutamatergic neurons from mouse embryonic stem cells requires raptor S6K signaling
AU - Chuang, Jen Hua
AU - Tung, Li Chu
AU - Yin, Yi
AU - Lin, Yenshou
N1 - Funding Information:
This work was partially supported by a grant ( NSC98-2311-B-003-MY3 and NSC101-2311-B-003-005 ) to Y. Lin from the National Science Council, Taiwan . We are grateful for the help from the Molecular Imaging Core Facility of National Taiwan Normal University and National RNAi Core Facility Platform of Academia Sinica (Taipei, Taiwan). We would like to thank Drs. Joseph Avruch and Sara Ortiz-Vega of Massachusetts General Hospital (Boston, MA, USA) for the critical reading of the manuscript and technical assistance.
PY - 2013/11
Y1 - 2013/11
N2 - Although the mammalian target of rapamycin complex 1 (mTORC1) functions as an important signaling complex in many cellular processes, the role of mTORC1 in neurons derived from embryonic stem cells (ESCs) has been less explored. Here, using a modified protocol to differentiate mouse ESCs (mESCs) into almost uniform glutamatergic neurons, we explored the importance of raptor/mTORC1 in the differentiation of mESCs. Raptor gene-trap mESCs, and raptor-knockdown mESCs formed smaller-sized embryonic bodies than the wild type and failed to undergo neuronal differentiation. Treatment with 1. μM rapamycin starting at the point when neuronal precursors began to differentiate from mESCs caused the gradual loss of neurites, shrinkage of soma, and a decreased ratio of neurite length to cell number over 48 to 72. h of treatment. This change was accompanied by activation of caspase-3 and S6 kinase (S6K), but not 4E-binding protein 1 (4EBP1). Knockdown of raptor during neuronal differentiation from mESCs also resulted in gradual loss of neurites and shrinkage of cell bodies. Loss of neurite density resulting from rapamycin treatment could be reversed by overexpression of S6K T389E. Taken together, these data demonstrate that raptor/mTORC1/S6K plays a critical role in the differentiation and survival of neurons derived from mESCs.
AB - Although the mammalian target of rapamycin complex 1 (mTORC1) functions as an important signaling complex in many cellular processes, the role of mTORC1 in neurons derived from embryonic stem cells (ESCs) has been less explored. Here, using a modified protocol to differentiate mouse ESCs (mESCs) into almost uniform glutamatergic neurons, we explored the importance of raptor/mTORC1 in the differentiation of mESCs. Raptor gene-trap mESCs, and raptor-knockdown mESCs formed smaller-sized embryonic bodies than the wild type and failed to undergo neuronal differentiation. Treatment with 1. μM rapamycin starting at the point when neuronal precursors began to differentiate from mESCs caused the gradual loss of neurites, shrinkage of soma, and a decreased ratio of neurite length to cell number over 48 to 72. h of treatment. This change was accompanied by activation of caspase-3 and S6 kinase (S6K), but not 4E-binding protein 1 (4EBP1). Knockdown of raptor during neuronal differentiation from mESCs also resulted in gradual loss of neurites and shrinkage of cell bodies. Loss of neurite density resulting from rapamycin treatment could be reversed by overexpression of S6K T389E. Taken together, these data demonstrate that raptor/mTORC1/S6K plays a critical role in the differentiation and survival of neurons derived from mESCs.
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U2 - 10.1016/j.scr.2013.08.003
DO - 10.1016/j.scr.2013.08.003
M3 - Article
C2 - 23988668
AN - SCOPUS:84883253900
SN - 1873-5061
VL - 11
SP - 1117
EP - 1128
JO - Stem Cell Research
JF - Stem Cell Research
IS - 3
ER -