We investigated the role of maternal exposure to human influenza virus (HI) in C57BL/6 mice on day 9 of pregnancy on hippocampal expression of nNOS in day 0 neonates and compared that to sham-infected pups. Qualitative analysis using polyclonal antibody to nNOS showed overall increases in immunoreactivity (IR) in hippocampal and dentate layers of day 0 infected neonates when compared to sham-infected animals. These increases in nNOS immunoreactivity were pronounced in hippocampal plate, intermediate, molecular, subplate, and dentate areas. Quantitative analysis of specific immunogold silver-enhanced nNOS IR via densitometry showed nNOS IR increases of 26-71.6% in all layers, i.e., hippocampal plate (35.1%), dentate area (71.6%), molecular area (43.75%), subplate (45.7%), and intermediate zone (26%) in infected neonatal brains vs. controls. The changes in levels of nNOS expression in hippocampi of neonates born to mothers exposed to HI virus during the second trimester of pregnancy may reflect the potential for glutamatergic excitotoxicity via activation of NMDA receptors in the developing brains of these neonatal mice.