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.
Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institute, Stockholm, Sweden.
Clas B
CB
Johansson
Carina
C
Lothian
Magnus
M
Molin
Hideyuki
H
Okano
Urban
U
Lendahl
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