PMU-Autor/inn/en
Aigner LudwigAbstract
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.
Useful keywords (using NLM MeSH Indexing)
5" Flanking Region/genetics
Animals
Base Sequence
Blotting, Western
Brain/cytology
Brain/embryology
Brain/metabolism
Cell Lineage/genetics
Cells, Cultured
Gene Expression Regulation/drug effects
Growth Substances/pharmacology
Hippocampus/cytology
Humans
Mice
Mice, Inbred C57BL
Microtubule-Associated Proteins/biosynthesis
Microtubule-Associated Proteins/genetics*
Molecular Sequence Data
Neurons/metabolism*
Neuropeptides/biosynthesis
Neuropeptides/genetics*
Regulatory Sequences, Nucleic Acid/drug effects
Regulatory Sequences, Nucleic Acid/genetics*
Sequence Analysis, DNA
Sequence Deletion
Stem Cells/drug effects
Stem Cells/metabolism*
Transfection
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gene regulation