EGF-responsive rat neural stem cells: molecular follow-up of neuron and astrocyte differentiation in vitro.
Neural stem cells (NSCs) could be very useful for the "cell therapy" treatment of neurological disorders. For this reason basic studies aiming to well characterize the biology of NSCs are of great interest. We carried out a molecular and immunocytochemical analysis of EGF-responsive NSCs obtained from rat pups. After the initial growth of NSCs as floating neurospheres in EGF-containing medium, cells were plated on poly-L-ornithine-coated dishes either in the presence or absence of EGF. We followed cell differentiation and apoptosis for 21 days in vitro and analyzed the expression levels of some genes having a major role in these processes, such as pRB, pRB2/p130, p27, and p53. We observed that EGF impairs neuronal differentiation. Furthermore, in the absence of mitogens, apoptosis, which appeared to proceed through the "p53 network," was significantly lower than in the presence of EGF. The cyclin kinase inhibitor p27, while important for cell cycle exit, seemed dispensable for cell survival and differentiation.
Jori
F P
FP
Department of Neurological Sciences, Second University of Naples, Naples, Italy.
Galderisi
U
U
Piegari
E
E
Cipollaro
M
M
Cascino
A
A
Peluso
G
G
Cotrufo
R
R
Giordano
A
A
Melone
M A B
MA
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001798:Blood Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D018797:Cell Cycle Proteins / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D019070:Cell Lineage / Q000187:drug effects* / Q000502:physiology; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D016160:Retinoblastoma Protein / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D050717:Retinoblastoma-Like Protein p130; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000187:drug effects / Q000378:metabolism
D001798:Blood Proteins; C493962:Cdkn1b protein, rat; D018797:Cell Cycle Proteins; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D011506:Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D016160:Retinoblastoma Protein; D050717:Retinoblastoma-Like Protein p130; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004815:Epidermal Growth Factor