TY - JOUR
T1 - Superoxide production in rat hippocampal neurons
T2 - Selective imaging with hydroethidine
AU - Bindokas, Vytautas P.
AU - Jordán, Joaquín
AU - Lee, Chong C.
AU - Miller, Richard J.
PY - 1996/2/15
Y1 - 1996/2/15
N2 - Digital-imaging microfluorimetry of the oxidation of hydroethidine (HEt) to ethidium can be used to monitor superoxide (O2) production selectively within individual rat hippocampal pyramidal neurons in culture and in brain slices. Under assay conditions, oxidation was not accomplished by hydroxyl radical, singlet O2, H2O2, or nitrogen radicals. Neuronal O2 production varied with metabolic activity and age. O2 generation increased after treatment with AMPA, kainic acid, and NMDA, and the mitochondrial uncoupler carbonylcyanide p-(trifluoromethoxy)phenyl hydrazone, but usually not after depolarization (50 mM K+). O2 concentrations were sensitive to scavengers and nitric oxide. HEt oxidation was higher in Ca2+-containing versus Ca2+-free saline. However, Ca2+ ionophores did not increase oxidation greatly. H2O2 application produced a secondary increase in O2. The major source of O2 under basal and stimulated conditions appeared to be the mitochondria. Consistent with this, ethidium staining in dendrites was punctate, colocalized with mitochondria, and blocked by CN.
AB - Digital-imaging microfluorimetry of the oxidation of hydroethidine (HEt) to ethidium can be used to monitor superoxide (O2) production selectively within individual rat hippocampal pyramidal neurons in culture and in brain slices. Under assay conditions, oxidation was not accomplished by hydroxyl radical, singlet O2, H2O2, or nitrogen radicals. Neuronal O2 production varied with metabolic activity and age. O2 generation increased after treatment with AMPA, kainic acid, and NMDA, and the mitochondrial uncoupler carbonylcyanide p-(trifluoromethoxy)phenyl hydrazone, but usually not after depolarization (50 mM K+). O2 concentrations were sensitive to scavengers and nitric oxide. HEt oxidation was higher in Ca2+-containing versus Ca2+-free saline. However, Ca2+ ionophores did not increase oxidation greatly. H2O2 application produced a secondary increase in O2. The major source of O2 under basal and stimulated conditions appeared to be the mitochondria. Consistent with this, ethidium staining in dendrites was punctate, colocalized with mitochondria, and blocked by CN.
KW - HO
KW - NMDA
KW - NOS
KW - dihydro ethidium
KW - excitotoxicity
KW - free radicals
KW - microfluorimetry
KW - mitochondria
KW - reactive oxygen species
UR - http://www.scopus.com/inward/record.url?scp=0030008052&partnerID=8YFLogxK
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U2 - 10.1523/jneurosci.16-04-01324.1996
DO - 10.1523/jneurosci.16-04-01324.1996
M3 - Article
C2 - 8778284
AN - SCOPUS:0030008052
SN - 0270-6474
VL - 16
SP - 1324
EP - 1336
JO - Journal of Neuroscience
JF - Journal of Neuroscience
IS - 4
ER -