Naturally occurring cell death via apoptosis occurs in the substantia nigra pars compacta (SNc) during rat development, culminating during the perinatal period. We previously showed that lipid peroxidation-mediated oxidative stress is not involved in this cell death process. Nitric oxide (NO) has been proposed to be critical for many developmental processes in brain and has been shown to mediate cell death in neurotoxin models of neurodegenerative disorders. Here, we reported that in vivo pre- and postnatal treatment with the non-specific NO synthase (NOS) inhibitor, L-NAME (60 mg/kg), or with the neuronal NOS inhibitor, 7-NI (30 mg/kg), dramatically decreased the NOS activity as well as the NADPH-diaphorase staining in brain. However, those treatments did not rescue dopamine neurons from developmental death, suggesting that NO is not involved in vivo in developmental death of these neurons or in the overall development of the SNc.