Premium
Rhotekin modulates differentiation of cultured neural stem cells to neurons
Author(s) -
Iwai Takashi,
Saitoh Akiyoshi,
Yamada Misa,
Takahashi Kou,
Hashimoto Eri,
Ukai Wataru,
Saito Toshikazu,
Yamada Mitsuhiko
Publication year - 2012
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.23029
Subject(s) - neurite , neural stem cell , transfection , microbiology and biotechnology , immunocytochemistry , biology , cellular differentiation , cell growth , western blot , mtt assay , stem cell , neuron , cell culture , neuroscience , in vitro , biochemistry , endocrinology , gene , genetics
Rhotekin is a downstream signal of Rho and is expressed in the central nervous system. However, the physiological role of rhotekin in the development of neural stem cells (NSCs) into neurons is unknown. In this study, we knocked down the expression of rhotekin protein with small interfering RNA (siRNA) in the NSCs and in neural differentiated cells and measured cell proliferation, differentiation, neurite length, and survival. By using immunocytochemistry and Western blot, the production of rhotekin was observed in NSCs and neuronal cells. Furthermore, rhotekin production was increased in accordance with neural differentiation. Rhotekin knock‐down reduced 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) activity and increased the cell death 72 hr after transfection in neurons. On the other hand, in NSCs, rhotekin knock‐down increased MTT activity and the number of 5‐bromo‐2′‐deoxyuridine (BrdU)‐positive cells. In the present study, we demonstrated that rhotekin is required for maintenance and survival of neurons and positively regulates differentiation and neurite outgrowth. Moreover, we found that rhotekin is produced in NSCs and that the role of rhotekin is to regulate cell proliferation negatively. In conclusion, these results suggest that rhotekin is one of the key molecules in the differentiation of NSCs into neurons. © 2012 Wiley Periodicals, Inc.