title: Cyclic AMP differentiation of the oligodendroglial cell line N20.1 switches staurosporine-induced cell death from necrosis to apoptosis.
abstract: Understanding the regulation of cell death pathways is critical for protecting myelin-producing cells and their associated axons during injury resulting from multiple sclerosis and other degenerative diseases. The immortalized N20.1 oligodendroglial cell line provides a useful model for identifying mechanisms that can be exploited to attenuate cell death in myelin-producing cells and their precursors. In our hands, the N20.1 cell line exhibits different characteristics and morphology depending on temperature (permissive or non-permissive) and the presence of cAMP-elevating agents (Studzinski et al. [1998] Neurochem. Res. 23:435-441; Boullerne et al. [1999] J. Neurochem. 72:1050-1060; Studzinski et al. [1999] J. Neurosci. Res. 57:633-642). Our laboratory previously observed that NO donors cause primarily necrotic death in N20.1 cells grown at permissive temperature, but the NO donor SNP switched a portion of cell death to the apoptic pathway. We have continued our study of apoptotic death in these cells by comparing the effects of staurosporine, a known apoptotic agent, on cells grown at the permissive temperature ("undifferentiated") vs. the non-permissive temperature in the presence of forskolin ("differentiated"). Undifferentiated N20.1 cells exhibit maximal cell death after 24 hr of exposure to 50 nM staurosporine, whereas differentiated cells show delayed cell death, with maximal death seen after 48 hr. Pyknotic nuclei were observed in both growth conditions; however, differentiated cells were protected by caspase inhibitors, whereas undifferentiated cells were not. Increased ssDNA staining and DNA laddering were found following 24-hr staurosporine treatment in the differentiated cells only. These results support the conclusion that N20.1 cells can switch from necrotic to apoptotic cell death when cell division is slowed and cyclic AMP is elevated.
authors.affiliation: Department of Neurology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Studzinski
authors.forename: J A
authors.initials: JA
authors.lastname: Benjamins
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D002461:Cell Line, Transformed / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002467:Cell Nucleus / Q000187:drug effects / Q000473:pathology; D002490:Central Nervous System / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D004396:Coloring Agents; D000242:Cyclic AMP / Q000378:metabolism*; D053938:DNA Fragmentation / Q000187:drug effects / Q000502:physiology; D004277:DNA, Single-Stranded / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D009103:Multiple Sclerosis / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D009336:Necrosis; D009836:Oligodendroglia / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011419:Propidium; D019311:Staurosporine / Q000494:pharmacology*; D013696:Temperature; D014343:Trypan Blue
chemical_list: D061945:Caspase Inhibitors; D004396:Coloring Agents; D004277:DNA, Single-Stranded; D004791:Enzyme Inhibitors; D011419:Propidium; D000242:Cyclic AMP; D020169:Caspases; D019311:Staurosporine; D014343:Trypan Blue