Fluoride (F) and lead (Pb) are two common environmental pollutants which are linked to the lowered intelligence, especially for children. Glutamate, a major excitatory neurotransmitter in the central nervous system, plays an important role in the process of learning and memory. However, the impact of F and Pb alone or in combination on glutamate metabolism in brain is little known. The present study was conducted to assess the glutamate level and the activities of glutamate metabolism related enzymes including asparate aminotransferase (AST), alanine aminotransferase (ALT) and glutamic acid decarboxylase (GAD) in the hippocampus, as well as learning abilities of offspring rat pups at postnatal week 6, 8, 10 and 12 exposed to F and/or Pb. During lactation, the pups ingested F and/or Pb via the maternal milk, whose mothers were exposed to sodium fluoride (150 mg/L in drinking water) and/or lead acetate (300 mg/L in drinking water) from the day of delivery. After weaning at postnatal day 21, the pups were exposed to the same treatments as their mother. Results showed that the learning abilities and hippocampus glutamate levels were significantly decreased by F and Pb individually and the combined interaction of F and Pb. The activities of AST and ALT in treatment groups were significantly inhibited, while the activities of GAD were increased, especially in rats exposed to both F and Pb together. These findings suggested that alteration of hippocampus glutamate by F and/or Pb may in part reduce learning ability in rats.