| Popis: |
Moderate, transient oxidative stress is achieved in SH-SY5Y cells using tertiary butylhydroperoxide as oxidant. Over a recovery period of 24 h, the enzymatic activity and protein levels of the acetylcholinesterase (AChE) splice variants tailed AChE (AChE-T) and readthrough AChE (AChE-R) are monitored. Their time-dependent correlation to pro- and anti-apoptotic factors, namely caspase 3 and Bcl-2, respectively, as well as lactate dehydrogenase release as a measure of cell viability is assessed. A distinctly different expression pattern of AChE-T as compared with AChE-R is recorded, in that AChE-T shows only a very slight increase over a 6 h time period. In contrast, AChE-R rises continuously during the recovery period, reaching peak intracellular levels that are up to six times higher than control levels 3-4 h post-stress, and is released from cells in substantial amounts. Moreover, anti-apoptotic Bcl-2 increases significantly, peaking 2-3 h after this AChE-R peak has occurred. We believe this study presents the first work that demonstrates - without relying on techniques of over-expression - the time-dependent correlation between apoptotic processes and related rescue mechanisms involving AChE isoforms in a neuronal cell line. |
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