Attenuation of glutamate-induced neuronal swelling and toxicity in transgenic mice overexpressing human CuZn-superoxide dismutase.
The role of oxygen-derived free radicals, superoxide in particular, in the pathogenesis of neuronal cell death induced by glutamate was studied using primary culture cortical neurons from transgenic mice overexpressing human CuZn-superoxide dismutase. Primary cortical neuron cultures were developed form 15-day-old fetuses of both transgenic mice and their normal littermates. Both human CuZn-superoxide dismutase and host mouse CuZn-superoxide dismutase activities in cultured neurons were identified by native gel electrophoresis followed by nitroblue tetrazolium staining. Cultured neurons grown for 10-12 days in vitro were exposed briefly to 0.5 mM glutamate for 5 minutes, followed by biochemical and morphological examinations at 2 and 4 hours. Our data have demonstrated that glutamate neurotoxicity is significantly reduced in transgenic neurons at 2 and 4 hours following exposure to glutamate, as measured by the intracellular 3-0-methyl glucose space, the efflux of lactate dehydrogenase, and by phase-contrast and bright-field trypan blue staining. These data indicate that transgenic neurons containing two- to threefold the normal amount of CuZn-superoxide dismutase activity are protected against glutamate neurotoxicity in vitro. Our results suggest that oxidative stress play an important role in glutamate-induced neuronal swelling and toxicity.
Chan
P H
PH
Department of Neurosurgery, University of California School of Medicine, San Francisco.
Chu
L
L
Chen
S F
SF
Carlson
E J
EJ
Epstein
C J
CJ
D019325:3-O-Methylglucose; D000818:Animals; D005971:Glutamates / Q000494:pharmacology* / Q000506:poisoning; D018698:Glutamic Acid; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008757:Methylglucosides / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D014343:Trypan Blue
D005971:Glutamates; D008757:Methylglucosides; D019325:3-O-Methylglucose; D018698:Glutamic Acid; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase; D014343:Trypan Blue