Effects of peroxidation and aging on rat neocortical ACh-release and protein kinase C

Cerebral cortical synaptosomes were prepared from 2- or 24-month-old Fischer 344 rats and exposed to a peroxidizing condition (50 microM ferrous ions and 2 microM ascorbate ions) before measuring either the release of newly synthesized [3H]acetylcholine (ACh) or protein kinase C activity (PKC). Several secretagogues with different mechanisms of action and different responses to aging were used to trigger release: K+ depolarization (5 mM-60 mM), calcium ionophore A23187 (1-10 micrograms/ml), and 4-aminopyridine (0.1-10 mM). Aging reduced K+ depolarization-induced release at every K+ concentration studied, reduced A23187-induced release at low but not high concentrations and did not affect 4-aminopyridine-induced release. Membrane peroxidation of synaptosomes from 2-month-old rats altered the response to secretagogues to match that seen in 24-month-old rats. Membrane peroxidation also attenuated the A23187-stimulated translocation of free to bound synaptosomal PKC activity in 2-month-old but not 24-month-old animals. These results suggest that membrane peroxidation may mimic some age-related deficits in secretagogue-induced [3H]ACh release.