Neuronal precursor-specific activity of a human doublecortin regulatory sequence.
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.
Karl
Claudia
C
Volkswagen-Foundation-Research Group, University of Regensburg, Regensburg, Germany.
Couillard-Despres
Sebastien
S
Prang
Peter
P
Munding
Matthias
M
Kilb
Werner
W
Brigadski
Tanja
T
Plötz
Sonja
S
Mages
Wolfgang
W
Luhmann
Heiko
H
Winkler
Jürgen
J
Bogdahn
Ulrich
U
Aigner
Ludwig
L
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D001921:Brain / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D019070:Cell Lineage / Q000235:genetics; D002478:Cells, Cultured; D000089622:Doublecortin Domain Proteins; D000089662:Doublecortin Protein; D005786:Gene Expression Regulation / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism*; D009479:Neuropeptides / Q000096:biosynthesis / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000187:drug effects / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism*; D014162:Transfection
C000714328:DCX protein, human; C000714847:Dcx protein, mouse; D000089622:Doublecortin Domain Proteins; D000089662:Doublecortin Protein; D006133:Growth Substances; D008869:Microtubule-Associated Proteins; D009479:Neuropeptides