Nestin enhancer requirements for expression in normal and injured adult CNS.
The nestin gene is expressed in many CNS stem/progenitor cells, both in the embryo and the adult, and nestin is used commonly as a marker for these cells. In this report we analyze nestin enhancer requirements in the adult CNS, using transgenic mice carrying reporter genes linked to three different nestin enhancer constructs: the genomic rat nestin gene and 5 kb of upstream nestin sequence (NesPlacZ/3), 636 bp of the rat nestin second intron (E/nestin:EGFP), and a corresponding 714 bp region from the human second intron (Nes714tk/lacZ). NesPlacZ/3 and E/nestin:EGFP mice showed reporter gene expression in stem cell-containing regions of brain and spinal cord during normal conditions. NesPlacZ/3 and E/nestin:EGFP mice showed increased expression in spinal cord after injury and NesPlacZ/3 mice displayed elevated expression in the periventricular area of the brain after injury, which was not the case for the E/nestin:EGFP mice. In contrast, no expression in adult CNS in vivo was seen in the Nes714tk/lacZ mice carrying the human enhancer, neither during normal conditions nor after injury. The Nes714 tk/lacZ mice, however, expressed the reporter gene in reactive astrocytes and CNS stem cells cultured ex vivo. Collectively, this suggests a species difference for the nestin enhancer function in adult CNS and that elements outside the second intron enhancer are required for the full injury response in vivo.
Johansson
Clas B
CB
Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institute, Stockholm, Sweden.
Lothian
Carina
C
Molin
Magnus
M
Okano
Hideyuki
H
Lendahl
Urban
U
D000367:Age Factors; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology; D001930:Brain Injuries / Q000503:physiopathology*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D004815:Epidermal Growth Factor / Q000235:genetics; D005260:Female; D015870:Gene Expression / Q000502:physiology; D017930:Genes, Reporter; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics*; D007438:Introns / Q000235:genetics; D007763:Lac Operon; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D013119:Spinal Cord Injuries / Q000503:physiopathology*; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D014407:Tumor Cells, Cultured
D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; C577486:Nes protein, rat; D064231:Nestin; D004815:Epidermal Growth Factor