Couillard-Despres, Sebastien and Quehl, Eike and Altendorfer, Katrin and Karl, Claudia and Ploetz, Sonja and Bogdahn, Ulrich and Winkler, Juergen and Aigner, Ludwig (2008) Human in vitro reporter model of neuronal development and early differentiation processes. BMC NEUROSCIENCE, 9: 31. ISSN 1471-2202,
Full text not available from this repository. (Request a copy)Abstract
Background: During developmental and adult neurogenesis, doublecortin is an early neuronal marker expressed when neural stem cells assume a neuronal cell fate. To understand mechanisms involved in early processes of neuronal fate decision, we investigated cell lines for their capacity to induce expression of doublecortin upon neuronal differentiation and develop in vitro reporter models using doublecortin promoter sequences. Results: Among various cell lines investigated, the human teratocarcinoma cell line NTERA-2 was found to fulfill our criteria. Following induction of differentiation using retinoic acid treatment, we observed a 16-fold increase in doublecortin mRNA expression, as well as strong induction of doublecortin polypeptide expression. The acquisition of a neuronal precursor phenotype was also substantiated by the establishment of a multipolar neuronal morphology and expression of additional neuronal markers, such as Map2, beta III-tubulin and neuron-specific enolase. Moreover, stable transfection in NTERA-2 cells of reporter constructs encoding fluorescent or luminescent genes under the control of the doublecortin promoter allowed us to directly detect induction of neuronal differentiation in cell culture, such as following retinoic acid treatment or mouse Ngn2 transient overexpression. Conclusion: Induction of doublecortin expression in differentiating NTERA-2 cells suggests that these cells accurately recapitulate some of the very early events of neuronal determination. Hence, the use of reporter genes under the control of the doublecortin promoter in NTERA-2 cells will help us to investigate factors involved early in the course of neuronal differentiation processes. Moreover the ease to detect the induction of a neuronal program in this model will permit to perform high throughput screening for compounds acting on the early neuronal differentiation mechanisms.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | EXPRESSION; DOUBLECORTIN; NEUROGENESIS; CELLS; MIGRATION; GROWTH; BRAIN; CNS; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Neurologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 09 Nov 2020 09:52 |
| Last Modified: | 09 Nov 2020 09:52 |
| URI: | https://pred.uni-regensburg.de/id/eprint/31359 |
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