title: EAAC1 gene deletion alters zinc homeostasis and enhances cortical neuronal injury after transient cerebral ischemia in mice.
abstract: The excitatory amino acids glutamate and cysteine are actively transported into neurons from the extracellular space by the high affinity glutamate transporter EAAC1. The astrocyte glutamate transporters, GLT1 and GLAST, are the primary mediators of glutamate clearance. EAAC1 has a limited role in this function. However, uptake of cysteine into neurons via EAAC1 contributes to neuronal antioxidant function by providing cysteine substrate for glutathione synthesis. Mice in which the EAAC1 gene has been deleted were seen to have enhanced susceptibility to neuronal oxidative stress and developed brain atrophy and cognitive function decline with aging. The aim of the current study was to evaluate if EAAC1 confers protection against ischemic events. Young adult CD-1 wild-type or EAAC1(-/-) mice were subjected to 30 min of bilateral common carotid artery occlusion and evaluated for neuronal death and zinc translocation. The intensity of TSQ fluorescence in the cytoplasm of cortical neurons in the EAAC1(-/-) mice was significantly higher than wild-type mice, indicating that the cortical neurons of EAAC1(-/-) mice contain higher cytoplasmic concentrations of labile (or free) zinc. Zinc translocation into cortical neurons was also enhanced in EAAC1(-/-) mice. Three days after ischemia, Fluoro-Jade B staining revealed that EAAC1(-/-) mice had more than twice as many degenerating neurons as wild-type mice. N-acetylcysteine, a membrane-permeant cysteine pro-drug, normalized basal zinc levels, reduced TSQ (+) neurons and reduced ischemic neuronal death in the EAAC1(-/-) mice when delivered in a pre-treatment fashion. Taken together, this study implicates EAAC1-dependent cysteine uptake as an endogenous source of enhancing antioxidant function and zinc homeostasis in neurons in the ischemic brain.
authors.affiliation: Department of Physiology, Hallym University, School of Medicine, Chuncheon, Republic of Korea.
authors.forename: Bong Geom
authors.initials: BG
authors.lastname: Jang
authors.forename: Seok Joon
authors.initials: SJ
authors.lastname: Won
authors.forename: Jin Hee
authors.initials: JH
authors.lastname: Kim
authors.forename: Bo Young
authors.initials: BY
authors.lastname: Choi
authors.forename: Min Woo
authors.initials: MW
authors.lastname: Lee
authors.forename: Min
authors.initials: M
authors.lastname: Sohn
authors.forename: Hong Ki
authors.initials: HK
authors.lastname: Song
authors.forename: Sang Won
authors.initials: SW
authors.lastname: Suh
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D050580:Excitatory Amino Acid Transporter 3 / Q000235:genetics / Q000378:metabolism*; D002546:Ischemic Attack, Transient / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008954:Models, Biological; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
chemical_list: D050580:Excitatory Amino Acid Transporter 3; D017382:Reactive Oxygen Species; C493399:Slc1a1 protein, mouse; D015032:Zinc; D000111:Acetylcysteine