Research on Regulation Function ofγ-Secretase Inhibitor DAPT on the Differentiation of Neural Precursor Cell Line_Journal of Biomedical Engineering

Authors：

WANGQi ,  LIHedong

Development and Stem Cell Institute, West China Second University Hospital, Sichuan University, Chengdu 610041, China;

Corresponding author：

LIHedong, Email: hedongli2009@hotmail.com

Keywords：

DAPT; neural precursor cell line; neuron; differentiation

DOI：

10.7507/1001-5515.20150023

Video：

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Abstract Full text Figures/Tables Video References Cited by

This study aims to investigate the effect ofγ-Secretase Inhibitor DAPT, (N-[N-(3, 5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester), on the differentiation of neural precursor cells and the production of neurons in the neural precursor cell line GE6. GE6 was cultured in medium with 4μmol/L DAPT added as the experimental group and the untreated medium separately as the control group. After 4 days of differentiation, we carried out the following experiments. We used immuno-fluorescent staining to observe the ratio of Tuj1, GFAP and O4 positive cells. We also used qRT-PCR to detect the effect of the DAPT on Tuj1 and GFAP mRNA transcription in the GE6. The results of immuno-fluorescent staining indicated that the Tuj1 ratio of experimental group was higher compared to that of the control group, but the GFAP and O4 ratio of experimental group was lower than that of the control group. The differences were statistically significant (P < 0.05). The result of qRT-PCR was in accordance with immunofluorescent staining results. It was well concluded that DAPT could promote the neurogenic differentiation of neural precursor cell line rather than leading to gliogenic differentiation. More neurons could be obtained for transplantation with the addition of DAPT.

Citation： WANGQi, LIHedong. Research on Regulation Function ofγ-Secretase Inhibitor DAPT on the Differentiation of Neural Precursor Cell Line. Journal of Biomedical Engineering, 2015, 32(1): 126-130. doi: 10.7507/1001-5515.20150023 Copy

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10.	MIDDELDORP J, HOL E M. GFAP in health and disease[J]. Prog Neurobiol, 2011, 93(3): 421-443.
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12.	KONRADI C, ZIMMERMAN E I, YANG C K, et al. Hippocampal interneurons in bipolar disorder[J]. Arch Gen Psychiatry, 2011, 68(4): 340-350.
13.	ROPER S N, STEINDLER D A. Stem cells as a potential therapy for epilepsy[J]. Exp Neurol, 2013, 244: 59-66.
14.	ALVAREZ-DOLADO M, CALCAGNOTTO M E, KARKAR K M, et al. Cortical inhibition modified by embryonic neural precursors grafted into the postnatal brain[J]. J Neurosci, 2006, 26(28): 7380-7389.
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1. BREDESEN D E, RAO R V, MEHLEN P. Cell death in the nervous system[J]. Nature, 2006, 443(7113): 796-802.
2. PAVLOV I, HUUSKO N, DREXEL M, et al. Progressive loss of phasic, but not tonic, GABAA receptor-mediated inhibition in dentate granule cells in a model of post-traumatic epilepsy in rats[J]. Neuroscience, 2011, 194: 208-219.
3. Anderson S A, Baraban S C. Cell therapy using GABAergic neural progenitors[M]//NOEBELS J L, AVOLI M, ROGAWSKI M A, et al. Jasper's Basic Mechanisms of the Epilepsies [Internet]. 4th edition. Bethesda (MD): National Center for Biotechnology Information (US), 2012.
4. SVENDSEN C N, CALDWELL M A, SHEN Jinkun, et al. Long-term survival of human central nervous system progenitor cells transplanted into a rat model of Parkinson's disease[J]. Exp Neurol, 1997, 148(1): 135-146.
5. LINDVALL O, KOKAIA Z, MARTINEZ-SERRANO A. Stem cell therapy for human neurodegenerative disorders-how to make it work[J]. Nat Med, 2004, 10(Suppl): S42-S50.
6. WANG M M. Notch signaling and Notch signaling modifiers[J]. Int J Biochem Cell Biol, 2011, 43(11): 1550-1562.
7. GELING A, STEINER H, WILLEM M, et al. Aγ-secretase inhibitor blocks Notch signaling in vivo and causes a severe neurogenic phenotypein zebrafish[J]. EMBO Rep, 2002, 3(7): 688-694.
8. CRAWFORD T Q, ROELINK H. The notch response inhibitor DAPT enhances neuronal differentiation in embryonic stem cell-derived embryoid bodies independently of sonic hedgehog signaling[J]. Dev Dyn, 2007, 236(3): 886-892.
9. LI Hedong, HADER A T, HAN Y R, et al. Isolation of a novel rat neural progenitor clone that expresses Dlx family transcription factors and gives rise to functional GABAergic neurons in culture[J]. Dev Neurobiol, 2012, 72(6): 805-820.
10. MIDDELDORP J, HOL E M. GFAP in health and disease[J]. Prog Neurobiol, 2011, 93(3): 421-443.
11. PIERFELICE T, ALBERI L, GAIANO N. Notch in the vertebrate nervous system: an old dog with new tricks[J]. Neuron, 2011, 69(5): 840-855.
12. KONRADI C, ZIMMERMAN E I, YANG C K, et al. Hippocampal interneurons in bipolar disorder[J]. Arch Gen Psychiatry, 2011, 68(4): 340-350.
13. ROPER S N, STEINDLER D A. Stem cells as a potential therapy for epilepsy[J]. Exp Neurol, 2013, 244: 59-66.
14. ALVAREZ-DOLADO M, CALCAGNOTTO M E, KARKAR K M, et al. Cortical inhibition modified by embryonic neural precursors grafted into the postnatal brain[J]. J Neurosci, 2006, 26(28): 7380-7389.
15. HUNT R F, GIRSKIS K M, RUBENSTEIN J L, et al. GABA progenitors grafted into the adult epileptic brain control seizures and abnormal behavior[J]. Nat Neurosci, 2013, 16(6): 692-697.

Journal of Biomedical Engineering

Research on Regulation Function ofγ-Secretase Inhibitor DAPT on the Differentiation of Neural Precursor Cell Line

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