Human medulloblastoma cell line DEV is a potent tool to screen for factors influencing differentiation of neural stem cells.
The aim of our study was to investigate whether a human neural cell line could be used as a reliable screening tool to examine the functional conservation, in humans, of transcription factors involved in neuronal or glial specification in other species. Gain-of-function experiments were performed on DEV cells, a cell line derived from a human medulloblastoma. Genes encoding nine different transcription factors were tested for their influence on the process of specification of human DEV cells towards a neuronal or glial fate. In a first series of experiments, DEV cells were transfected with murine genes encoding transcription factors known to be involved in the neuronal differentiation cascade. Neurogenins-1, -2, and -3; Mash-1; and NeuroD increased the differentiation of DEV cells towards a neuronal phenotype by a factor of 2-3.5. In a second series of experiments, we tested transcription factors involved in invertebrate glial specification. In the embryonic Drosophila CNS, the development of most glial cells depends on the master regulatory gene glial cell missing (gcm). Expression of gcm in DEV cells induced a twofold increase of astrocytic and a sixfold increase of oligodendroglial cell types. Interestingly, expression of tramtrack69, which is required in all Drosophila glial cells, resulted in a fourfold increase of only the oligodendrocyte phenotype. Expression of the related tramtrack88 protein, which is not expressed in the fly glia, or the C. elegans lin26 protein showed no effect. These results show that the Drosophila transcription factor genes tested can conserve their function upon transfection into the human DEV cells, qualifying this cell line as a screening tool to analyze the mechanisms of neuronal and glial specification.
Buzanska
L
L
Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
Spassky
N
N
Belin
M F
MF
Giangrande
A
A
Guillemot
F
F
Klämbt
C
C
Labouesse
M
M
Thomas
J L
JL
Domanska-Janik
K
K
Zalc
B
B
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002528:Cerebellar Neoplasms; D003470:Culture Media / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics; D004330:Drosophila; D029721:Drosophila Proteins; D015870:Gene Expression / Q000502:physiology; D049452:Green Fluorescent Proteins; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D008527:Medulloblastoma; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology*; D009479:Neuropeptides / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D013234:Stem Cells / Q000166:cytology*; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D051792:Basic Helix-Loop-Helix Proteins; D029742:Caenorhabditis elegans Proteins; D003470:Culture Media; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C065500:ASCL1 protein, human; C102805:GCM protein, Drosophila; C105389:NEUROG1 protein, human; C491183:NEUROG2 protein, human; C498695:NEUROG3 protein, human; C090507:lin-26 protein, C elegans; C075878:ttk protein, Drosophila; C093564:Neurogenic differentiation factor 1; C486336:Neurog1 protein, mouse