title: Dramatic increase in readthrough acetylcholinesterase in a cellular model of oxidative stress.
abstract: 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.
authors.affiliation: Department of Pharmacology, School of Pharmacy, Goethe University Frankfurt, Frankfurt am Main, Germany.
authors.forename: Ricarda
authors.initials: R
authors.lastname: Härtl
authors.forename: Anne
authors.initials: A
authors.lastname: Gleinich
authors.forename: Martina
authors.initials: M
authors.lastname: Zimmermann
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics / Q000378:metabolism*; D000704:Analysis of Variance; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008970:Molecular Weight; D009447:Neuroblastoma; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013997:Time Factors; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D020122:tert-Butylhydroperoxide; D007770:L-Lactate Dehydrogenase; D000110:Acetylcholinesterase; D053148:Caspase 3