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.