Base excision repair activities in organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation.
The capacity for DNA repair is likely to be one of the factors that determine the vulnerability of neurons to ischemic stress and may influence the pathological outcome of stroke. In this report, initiation of base excision repair (BER) was assessed by analysis of enzyme activity and gene expression level of DNA glycosylases and AP-endonucleases in rat organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation (OGD) - an in vitro model of stroke. Under basal conditions, AP-endonuclease activity and base removal of ethenoadenine and 8-oxoguanine (8-oxoG) were higher (by approximately 20-35 %) in CA3/fascia dentata (FD) than in CA1. Base removal of uracil did not differ between the two hippocampal regions, while removal of 5-hydroxyuracil (5-OHU) was slightly less efficient in CA3/FD than in CA1. Analyses performed immediately after 30 min of OGD revealed a decreased AP-endonuclease activity (by approximately 20%) in CA1 as well as CA3/FD, and an increased ethenoadenine activity (by approximately 25%) in CA1. Activities for 8-oxoG, 5-OHU and uracil showed no significant changes at this time point. At 8h after OGD, none of the enzyme activities differed from control values. Real-time RT-PCR showed that transcription of DNA glycosylases, including Ogg1, Nth1, Ung, Aag, Neil1 and Neil2 were not changed in response to OGD treatment (t=0 h). The hippocampal expression of Neil2 was low compared with the other DNA glycosylases. These data indicate that CA1 has a lower capacity than CA3/FD for removal of base lesions under basal conditions. The relatively low capacity for BER in basal conditions and the apparent failure to upregulate repair of oxidative damage after OGD might contribute to the high vulnerability of CA1 to ischemic injury.
Rolseth
Veslemøy
V
Centre for Molecular Biology and Neuroscience, Institute of Medical Microbiology, University of Oslo, Rikshospitalet, N-0027 Oslo, Norway.
Rundén-Pran
Elise
E
Neurauter
Christine Gran
CG
Yndestad
Arne
A
Luna
Luisa
L
Aukrust
Pål
P
Ottersen
Ole Petter
OP
Bjørås
Magnar
M
D000818:Animals; D001483:Base Sequence; D016923:Cell Death; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers; D004260:DNA Repair; D005947:Glucose / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D009924:Organ Culture Techniques; D010100:Oxygen / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D012333:RNA, Messenger; D045647:DNA Glycosylases; D005947:Glucose; D010100:Oxygen