Neuronal differentiation of PC12 cells as a result of prevention of cell death by bcl-2.
Rat pheochromocytoma PC12 cells die when cultured in serum-free medium. Neurotrophic factors can rescue PC12 cells from cell death, and induce neuronal differentiation. To further investigate the relationship among cell death, survival, and differentiation, the bcl-2 cDNA, which is known to prevent apoptosis in various types of cells, was transfected into PC12 cells. Six monoclonal bcl-2-transfected cell lines were isolated and confirmed to express mRNA and protein product of bcl-2. The wild-type and bcl-2-transfected PC12 cells were kept to adhere to collagen-coated dishes at the initiation of serum-free experiments to avoid cellular damage due to detachment of the cells by trituration. Even under the conditions, the control PC12 cells mostly died within 24 h, when cultured in serum-free medium, whereas those expressing Bcl-2 survived even for 7 days in serum-free medium. Moreover, outgrowth of long processes in the bcl-2-transfected cells was only observed under the condition to keep the cells attached to the dishes in serum-free medium without any additive neurotrophic or growth factors. Neurofilament medium protein, which is a neuron-specific cytoskeletal component, was also expressed in the differentiated cells, suggesting that the long processes in bcl-2-transfected PC12 cells are neurites. However, neuronal differentiation of PC12 cells expressing Bcl-2 was not observed when cultured in serum-containing medium. Accordingly, survival of PC12 cells expressing Bcl-2 under the condition which cells usually die may be accompanied with neuronal differentiation.
Sato
N
N
Department of Cell Biology and Neuroanatomy, Iwate Medical University, School of Medicine, Japan.
Hotta
K
K
Waguri
S
S
Nitatori
T
T
Tohyama
K
K
Tsujimoto
Y
Y
Uchiyama
Y
Y
D000818:Animals; D016923:Cell Death; D002454:Cell Differentiation; D016895:Culture Media, Serum-Free; D009474:Neurons / Q000166:cytology*; D016716:PC12 Cells / Q000378:metabolism* / Q000473:pathology*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2; D051381:Rats; D014162:Transfection
D016895:Culture Media, Serum-Free; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2