(+)-Cholesten-3-one induces differentiation of neural stem cells into dopaminergic neurons through BMP signaling.
To identify small molecules that induce dopaminergic neurons from neural stem cells (NSCs) is promising for therapy of Parkinson's disease. Here we report the results of analyzing structurally related steroids in traditional Chinese medicine to identify agents that enhance dopaminergic differentiation of NSCs. Using P19 cells transfected by tyrosine hydroxylase (TH) promoter reporter construct, (+)-Cholesten-3-one with carbonyl, but not cholesterol and cholesterol myristate can effectively promote the activity of TH promoter. This effect depends on bone morphogenetic protein (BMP) signaling. Phenotypic cellular analysis indicated that (+)-Cholesten-3-one induces differentiation of NSCs to dopaminergic neurons with increased expression of specific dopaminergic markers including TH, dopamine transporter, dopa decarboxylase and higher level of dopamine secretion. (+)-Cholesten-3-one significantly increases the expression of BMPR IB, but not BMPR IA or BMPR II; p-Smad1/5/8 positive nuclei and expression of p-Smad1/5/8 were detected in NSCs treated with (+)-Cholesten-3-one, indicating that (+)-Cholesten-3-one may activate the BMP signaling. Moreover, overexpression of BMP4 or inhibition of BMP affects the effect of (+)-Cholesten-3-one on the dopaminergic phenotype. These findings may contribute to efficient production of dopaminergic neurons from NSCs culture for many applications and raise interesting questions about the role of (+)-Cholesten-3-one in neurogenesis.
Chen
Dong-Feng
DF
The State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing, China. CDF27212@21cn.com
Meng
Ling-Jie
LJ
Du
Shao-Hui
SH
Zhang
Hai-Ling
HL
Li
Hui
H
Zhou
Jian-Hong
JH
Li
Yi-Wei
YW
Zeng
He-Ping
HP
Hua
Zi-Chun
ZC
D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D002783:Cholestenones / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000378:metabolism*; D005434:Flow Cytometry; D007150:Immunohistochemistry; D058953:Neural Stem Cells / Q000502:physiology*; D055495:Neurogenesis / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D019485:Bone Morphogenetic Proteins; D002783:Cholestenones; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine