Neuronal precursor-specific activity of a human doublecortin regulatory sequence
| Autor: | Tanja Brigadski, Werner Kilb, Heiko J. Luhmann, Ludwig Aigner, Jürgen Winkler, Wolfgang Mages, Peter Prang, Matthias Munding, Sebastien Couillard-Despres, Sonja Plötz, Claudia Karl, Ulrich Bogdahn |
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| Rok vydání: | 2005 |
| Předmět: |
Doublecortin Domain Proteins
Doublecortin Protein 5' Flanking Region Blotting Western Molecular Sequence Data Regulatory Sequences Nucleic Acid Transfection Biochemistry Hippocampus Cellular and Molecular Neuroscience Mice Gene expression Animals Humans Cell Lineage Growth Substances Gene Transcription factor Cells Cultured Sequence Deletion Regulation of gene expression Neurons Reporter gene biology Base Sequence Stem Cells Neurogenesis Neuropeptides Brain Sequence Analysis DNA Molecular biology Doublecortin Mice Inbred C57BL Gene Expression Regulation Regulatory sequence biology.protein Microtubule-Associated Proteins |
| Zdroj: | Journal of neurochemistry. 92(2) |
| ISSN: | 0022-3042 |
| Popis: | The doublecortin (DCX) gene encodes a 40-kDa microtubule-associated protein specifically expressed in neuronal precursors of the developing and adult CNS. Due to its specific expression pattern, attention was drawn to DCX as a marker for neuronal precursors and neurogenesis, thereby underscoring the importance of its promoter identification and promoter analysis. Here, we analysed the human DCX regulatory sequence and confined it to a 3.5-kb fragment upstream of the ATG start codon. We demonstrate by transient transfection experiments that this fragment is sufficient and specific to drive expression of reporter genes in embryonic and adult neuronal precursors. The activity of this regulatory fragment overlapped with the expression of endogenous DCX and with the young neuronal markers class III beta-tubulin isotype and microtubule-associated protein Map2ab but not with glial or oligodendroglial markers. Electrophysiological data further confirmed the immature neuronal nature of these cells. Deletions within the 3.5-kb region demonstrated the relevance of specific regions containing transcription factor-binding sites. Moreover, application of neurogenesis-related growth factors in the neuronal precursor cultures suggested the lack of direct signalling of these factors on the DCX promoter construct. |
| Databáze: | OpenAIRE |
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