Nestin expression is lost in a neural stem cell line through a mechanism involving the proteasome and Notch signalling.
Neural stem cells (NSCs) are believed to repair brain damage primarily through cell replacement: i.e., the ability to regenerate lost neurons and glia in a site-specific fashion. The neural stem cell line, MHP36, has been shown to have this capacity, but we have little idea of the molecular mechanisms that control the differentiation of such cells during brain repair. In this study we show that an early event in the differentiation of MHP36 cells, both in vivo and in vitro, is the loss of expression of the intermediate filament protein, nestin. We use a co-culture assay to show that loss of nestin is fast, being detectable after just 1 h and complete in 4 h, and is controlled by proteasome degradation rather than down-regulation of de novo nestin synthesis. We also show that nestin loss is regulated by Notch, and mediated by cell contact.
Mellodew
Karen
K
Department of Neuroscience, Institute of Psychiatry, PO39, 1 Windsor Walk, Denmark Hill, London, UK.
Suhr
Ralf
R
Uwanogho
Dafe A
DA
Reuter
Iris
I
Lendahl
Urban
U
Hodges
Helen
H
Price
Jack
J
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D018920:Coculture Techniques / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004622:Embryo, Mammalian; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D009097:Multienzyme Complexes / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D064231:Nestin; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D009930:Organic Chemicals; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D051880:Receptors, Notch; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors; D014162:Transfection / Q000379:methods
D015853:Cysteine Proteinase Inhibitors; D005456:Fluorescent Dyes; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D009097:Multienzyme Complexes; C489428:Myc protein, mouse; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; C577486:Nes protein, rat; D064231:Nestin; D009930:Organic Chemicals; C451241:PKH67; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D051880:Receptors, Notch; C067713:lactacystin; D049452:Green Fluorescent Proteins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D010751:Phosphopyruvate Hydratase; D000111:Acetylcysteine