Extending the protective recovery method:

Glutathione depletion underlies rapid deterioration of adult brain slices


     We investigated the integrity of brain slices prepared from older Thy1-GCaMP3 transgenic mice using Ca imaging and stimulation of neurons with application of 20-50 uM NMDA.  This agonist produced a more random activation of neurons, which was useful to see the distribution of healthy neurons in slices with a more moderate stimulation compared to high K+.  Brain slices were prepared from 13-19 month old mice using the choline-HEPES aCSF recovery method.  The control group had no treatment while the experimental group had either GSH-EE or NAC added to the slices.  Control slices were functional at early times but deteriorated after several hours.  Slices treated with NAC or GSH-EE remained functional with a healthy appearance for extended times out to 7+ hours after slicing.