Free radicals and aging. Electron spin resonance studies on neuronal lipopigments and cells grown in vitro

Abstract

Electron spin resonance (ESR) spectroscopy has been used in an attempt to observe free radicals which may be connected to the aging processes. Isolated lipofuscin particles from the human brain, which may act as traps for reactive ESR centers, have been studied. The lipopigments exhibit ESR signals which were attributed to metal ions. Both high-spin ferric iron and copper (II) complexes were observed. The copper resonance showed axial symmetry and the observed ESR parameters ( A ‖ = 185 gauss, g ‖ = 2.25 and g ⊥ = 2.066) indicate that the copper ions are of the “non-blue” (type (so-called “type 2”). The data suggest that the copper ions might be ligated to two nitrogen and two oxygen atoms. Ceroid particles from the brains of dogs with end-stage “canine ceroid lipofuscinosis” yield similar resonances involving metal complexes. The concentration of the copper ions in the lipopigments seems to increase slightly towards the end stage of life. Retinal pigment epithelial cells from the eyes of dogs with canine ceroid lipofuscinosis were grown in vitro and examined with ESR when the cells reached confluency and then after being maintained confluent for 2 weeks. The spectra indicate an accumulation of metal ions due to this “aging”. Phase-contrast microscopy reveals an increase in perinuclear dense bodies during the time in confluency. A weak ESR signal, compatible with an “oxygen-centered” radical, was also found to increase slightly during the “aging” period.