The DNA repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) is a cardinal defense against the mutagenic and carcinogenic effects of alkylating agents. We have reported evidence that absence of detectable MGMT activity (MGMT(-) phenotype) in human brain is a predisposing factor for primary brain tumors that affects ca. 12% of individuals [J.R. Silber, A. Blank, M.S. Bobola, B.A. Mueller, D.D. Kolstoe, G.A. Ojemann, M.S. Berger, Lack of the DNA repair protein O(6)-methylguanine-DNA methyltransferase in histologically normal brain adjacent to primary brain tumors, Proc. Natl. Acad. Sci. U.S.A. 93 (1996) 6941-6946]. We report here that MGMT(-) phenotype in the brain of children and adults, and the apparent increase in risk of neurocarcinogenesis, may arise during gestation. We found that MGMT activity in 71 brain specimens at 6-19 weeks post-conception was positively correlated with gestational age (P<or=0.0015). Moreover, the proportion of specimens exhibiting MGMT(-) phenotype (MGMT content<0.42 fmol/10(6)cells or 255 molecules/cell) declined progressively from 76% (16/21) at 6-8 weeks to 13% (1/8) at 15-19 weeks. All liver specimens that accompanied MGMT(-) brain (15/15) had measurable MGMT activity, demonstrating that the phenotype was not systemic in these cases. In contrast to MGMT, apurinic endonuclease, DNA polymerase beta and lactate dehydrogenase activities were found in every brain extract assayed, and showed no significant relationship with gestational age. The observed gestational pattern has at least two implications for neurocarcinogenesis. (1) Early in development, brain tissue that has MGMT(-) phenotype and is rapidly proliferating may be especially vulnerable to alkylation-induced mutations, including mutations that lead to brain tumors. (2) Persistence of prenatal MGMT deficiency into postnatal life in a sub-population of individuals may increase brain tumor risk. Our findings provide possible mechanistic insight into epidemiologic data associating maternal alkylating agent exposure with brain tumor incidence.