Intracerebroventricular administration of L-buthionine sulfoximine: a method for depleting brain glutathione

Sprague-Dawley rats (200-260 g) were anesthetized with chloral hydrate (400 mg/kg) and polyethylene cannulae were permanently implanted into the lateral ventricles. One or two days later, L-buthionine-[S,R]-sulfoximine (L-BSO), an apparently selective inhibitor of gamma-glutamylcysteine synthetase, was administered intracerebroventricularly through the cannulae. The brain content of glutathione (GSH) was determined by HPLC with electrochemical detection (gold/mercury electrode) using N-acetylcysteine as internal standard. A time-course study of the changes in the striatum following a single dose of L-BSO (3.2 mg) revealed a maximal depletion of GSH (-60%) approximately 48 h after the administration. The effects of various doses of L-BSO on GSH in the striatum, in the limbic region, and in the cortex were assessed at 24 h and 48 h after the administration. L-BSO (0.02-3.2 mg) produced dose-dependent reductions of GSH in all brain regions studied at both time intervals. In a long-term experiment L-BSO (3.2 mg) was administered every second day. After 4 days, i.e., after two injections, striatal GSH was reduced by approximately 70%. No further depletion of GSH was obtained by additional injections of L-BSO, but GSH was maintained at this low level for the 12 days studied. These results suggest that L-BSO, administered intracerebroventricularly, would serve as a useful tool for evaluation of the biological role of GSH in the CNS.