Glial cell-specific differences in response to alkylation damage.
Oligodendrocytes are preferentially sensitive to the toxic, carcinogenic, and teratogenic effects of methylnitrosourea (MNU). The mechanisms responsible for this enhanced sensitivity have not been fully elucidated. One of the most vulnerable cellular targets for this chemical is mitochondrial DNA (mtDNA). To determine if differences in mtDNA damage and repair capacity exist among the different CNS glial cell types, the effects of MNU exposure on oligodendroglia, astroglia, and microglia cultured separately from neonatal rat brain were compared. Quantitative determinations of mtDNA initial break frequencies and repair efficiencies showed that whereas no cell type-specific differences in initial mtDNA damage were detected, mtDNA repair in oligodendrocytes, oligodendrocyte progenitors, and microglia was significantly reduced compared to that of astrocytes. In astrocytes, and all other cell types previously evaluated in our laboratory, >60% of N-methylpurines were removed from the mtDNA by 24 hr. In contrast, only 35% of lesions were removed from mtDNA of oligodendrocytes, oligodendrocyte progenitors, and microglia during the same time period. Mitochondrial perturbations by a variety of xenobiotics have been linked to apoptosis. In the present study, apoptosis, as determined by DNA laddering and ultrastructural analysis, was clearly induced by MNU treatment of cultured oligodendrocyte progenitors and microglia, but not in astroglia. These data demonstrate a correlation between diminished mtDNA repair capacity and the induction of apoptosis. However, further experimentation is necessary to determine if a causal relationship exists and contributes to the vulnerability of oligodendroglia following exposure to N-nitroso compounds in the environment or in chemotherapeutic regimen.
Department of Structural and Cellular Biology, University of South Alabama, Mobile 36688, USA.
S P
SP
Ledoux
C C
CC
Shen
V I
VI
Grishko
P A
PA
Fields
A L
AL
Gard
G L
GL
Wilson
D000477:Alkylating Agents / Q000633:toxicity*; D000478:Alkylation; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D004249:DNA Damage; D053938:DNA Fragmentation; D015342:DNA Probes; D004272:DNA, Mitochondrial / Q000187:drug effects* / Q000502:physiology; D008770:Methylnitrosourea / Q000633:toxicity*; D009457:Neuroglia / Q000502:physiology* / Q000648:ultrastructure; D009836:Oligodendroglia / Q000187:drug effects / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000187:drug effects / Q000648:ultrastructure
D000477:Alkylating Agents; D015342:DNA Probes; D004272:DNA, Mitochondrial; D008770:Methylnitrosourea