Toxic effects of acetylcholinesterase on neuronal and glial-like cells in vitro.
Acetylcholinesterase (AChE), the enzyme involved in the hydrolysis of the neurotransmitter acetylcholine, has been implicated in non-cholinergic actions which may play a role in neurodegenerative diseases such as Alzheimer's disease. To study the potential cytotoxicity of brain AChE, the effects of affinity purified AChE were analyzed on neuronal (Neuro 2a) and glial-like (B12) cells. LDH release and MTT reduction assays showed that AChE was toxic; the toxicity was dependent on the enzyme concentration, time of incubation and cellular density. The toxic effect of AChE was not related to its catalytic activity, since the anti-cholinesterase drug BW284C51 and heat inactivation were unable to block the effects of the enzyme. When cells were incubated at 4 degrees C, toxicity was completely blocked, in contrast to cells incubated at 37 degrees C. The presence of serum in the culture medium inhibited the toxic effects of AChE. Cytoplasmic shrinkage, condensation and fragmentation of nucleus as well as DNA strand breaks detected with the TUNEL technique indicated that apoptotic cell death is involved in the effect of AChE. Considering that we have previously shown that AChE promotes the assembly of beta-amyloid peptide into neurotoxic amyloid fibrils, it is conceivable that the neurotoxicity of AChE shown here may play a role in the neuronal degeneration observed in Alzheimer's disease.
Departamento de Biología Celular y Molecular, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
F H
FH
Calderón
R
R
von Bernhardi
G
G
De Ferrari
S
S
Luza
R
R
Aldunate
N C
NC
Inestrosa
D000110:Acetylcholinesterase / Q000378:metabolism / Q000633:toxicity*; D000544:Alzheimer Disease / Q000378:metabolism; D000818:Animals; D001710:Biotin; D001798:Blood Proteins / Q000494:pharmacology; D002417:Cattle; D002452:Cell Count; D003080:Cold Temperature; D053938:DNA Fragmentation; D003856:Deoxyuracil Nucleotides; D004789:Enzyme Activation / Q000502:physiology; D051379:Mice; D009447:Neuroblastoma; D016874:Neurofibrillary Tangles / Q000201:enzymology; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects*; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects
D001798:Blood Proteins; D003856:Deoxyuracil Nucleotides; D001710:Biotin; D000110:Acetylcholinesterase