Neuroscience 2003 Abstract
| Presentation Number: | 672.7 |
|---|---|
| Abstract Title: | Differentiation of mesencephalic neural progenitors: effect of extracellular matrix and brain derived neurotrophic factor. |
| Authors: |
Takeshima, T.*1
; Fukuhara, Y.1
; Kashiwaya, Y.1,2
; Nakaso, K.1
; Wakutani, Y.1
; Nakashima, K.1
1Dept. of Neurol., Tottori Univ. Fac. of Med., Yonago, Japan 2MD, 36-1 Nishimachi, 683-8504, |
| Primary Theme and Topics |
Development - Neurogenesis and Gliogenesis -- Neural stem cells |
| Session: |
672. Midbrain Stem Cells Poster |
| Presentation Time: | Tuesday, November 11, 2003 3:00 PM-4:00 PM |
| Location: | Morial Convention Center - Hall F-I, Board # A36 |
| Keywords: | Parkinson's disease, DOPAMINE, nurr-1 |
We have reported a mesencephalic microisland culture method (J Neurosci 1994) and expansion of dopaminergic neurons with microisland culture (SFN Abstr. 1997;23:593). We have also established a protocol for robust expansion and cryopreservation of mesencephalic neural progenitors (MNP) from E14 rat brain. Here, we explored efficient differentiation protocols for the MNPs to dopaminergic neurons. RT-PCR expression analyses of nurr-1, shh, en-1, en-2, smo, ptc, pax-2, pax-8 were carried out in MNP cultures on various extracellular matrix (ECM), i.e. glass, poly-D- lysine (PDL), collagen, laminin, or fibronectin. Nurr-1, an essential transcription factor for dopaminergic neurons, was expressed in MNPs. The strongest expression of nurr-1 was observed in the cells grown on fibronectin. We tested brain derived neurotrophic factor (BDNF), forskolin, dopamine and ascorbic acid in various concentrations and in some combinations. The differentiated MNPs are evaluated at 3, 5, and 7 days in culture by immunostaing for microtubules associated protein-2 (MAP-2) and tyrosine hydroxylase (TH). We found a combination of 50ng/ml BDNF, 10µM dopamine, and 10µM forskolin increased the efficiency of differentiation of MNPs to dopaminergic neurons in serum-free cultures on fibronectin as ECM. This culture technique will provide new experimental model of dopaminergic neurons relevant to Parkinson's disease.
Supported by Min. Edu., Culture, Sports, Sci & Technology, Gov.Japan
Sample Citation:
[Authors]. [Abstract Title]. Program No. XXX.XX. 2003 Neuroscience Meeting Planner. New Orleans, LA: Society for Neuroscience, 2003. Online.
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