A characteristic feature of several encephalopathies is preferential impairment of cholinergic neurons. Their particular susceptibility to cytotoxic insults may result from the fact that they utilise acetyl-CoA both for energy production and acetylcholine synthesis. In addition, phenotypic modifications of cholinergic neurons are likely to influence their susceptibility to specific harmful conditions. SN56 cholinergic cells were differentiated by the combination of dibutyryl cAMP and retinoic acid. Al and sodium nitroprusside (SNP, NO donor) exerted direct additive inhibitory effects on mitochondrial aconitase activity. However, NO, Al, or amyloid beta (Abeta)(25-35) caused none or only slight changes of choline O-acetyl transferase (ChAT) and pyruvate dehydrogenase (PDH) activity and relatively small loss of non-differentiated cells (NCs). On the other hand, in differentiated cells (DCs) these neurotoxins brought about marked decreases of these enzyme activities along with greater than in non-differentiated ones increase of cell-death rate. Abeta(35-25) had no effect on these cell parameters. NO and other compounds aggravated detrimental effect of each other particularly in differentiated cells. Thus, differential vulnerability of brain cholinergic neurons to various degenerative signals may result from their phenotype-dependent ratios of acetylcholine to acetyl-CoA synthesising capacities.