title: Glial cell-specific differences in repair of O6-methylguanine.
abstract: Normal and malignant cells of the oligodendrocyte lineage show increased sensitivity to alkylating agents compared to astrocytes. One of the most mutagenic DNA lesions formed following exposure to alkylating agents is O6-alkylguanine. To determine whether the increased sensitivity to nitrosoureas seen in oligodendrocytes is due to decreased repair capacity for O6-alkylguanine, removal of this lesion from DNA was assessed in primary cultures of rat oligodendrocytes, astrocytes, and microglia. Glial cells were exposed to 1 mM N-methyl-N-nitrosourea for 1 h and allowed 8 or 24 h for repair. Repair was evaluated using an immunoslot blot technique and a monoclonal antibody which recognizes O6-methylguanine (O6MeGua). Astrocytes removed O6MeGua more efficiently (approximately 80% in 24 h) than either oligodendrocytes (approximately 20%) or microglia (approximately 4%). Determination of O6-alkylguanine-DNA-alkyltransferase (AT) activity revealed that astrocytes contain 0.4 pmol/mg protein, which is average by comparison to other cell types. Both oligodendrocytes and microglia exhibited very low levels of AT (oligodendrocytes, 0.08; microglia, 0.01 pmol/mg protein). These data are the first to show that within different populations of glial cells, O6MeGua adduct removal is substantially reduced in both oligodendrocytes and microglia. Rapid removal of O6MeGua in astrocytes coupled with persistence of this mutagenic lesion in oligodendrocytes following exposure of the developing central nervous system to nitrosoureas could contribute to the observed formation of oligodendrogliomas. Inefficient removal of O6MeGua in oligodendrogliomas might also account for their response to chemotherapeutic regimens involving alkylating agents such as procarbazine, lomustine, and carmustine. The lack of repair of O6MeGua in microglia suggests that primary lymphomas of the central nervous system might be sensitive to treatment with alkylating drugs whose toxicity depends on repair of this adduct.
authors.affiliation: University of South Alabama, Mobile 36688, USA.
authors.forename: S P
authors.initials: SP
authors.lastname: LeDoux
authors.forename: B A
authors.initials: BA
authors.lastname: Williams
authors.forename: B S
authors.initials: BS
authors.lastname: Hollensworth
authors.forename: C
authors.initials: C
authors.lastname: Shen
authors.forename: J
authors.initials: J
authors.lastname: Thomale
authors.forename: M F
authors.initials: MF
authors.lastname: Rajewsky
authors.forename: T P
authors.initials: TP
authors.lastname: Brent
authors.forename: G L
authors.initials: GL
authors.lastname: Wilson
mesh_terms: D000477:Alkylating Agents / Q000633:toxicity; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D004249:DNA Damage; D004260:DNA Repair; D006147:Guanine / Q000031:analogs & derivatives* / Q000378:metabolism; D008770:Methylnitrosourea / Q000633:toxicity; D008780:Methyltransferases / Q000378:metabolism; D017628:Microglia / Q000187:drug effects / Q000378:metabolism*; D019853:O(6)-Methylguanine-DNA Methyltransferase; D009836:Oligodendroglia / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D013997:Time Factors
chemical_list: D000477:Alkylating Agents; D006147:Guanine; D008770:Methylnitrosourea; C008449:O-(6)-methylguanine; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase