Downregulation of Cu/Zn superoxide dismutase leads to cell death via the nitric oxide-peroxynitrite pathway

CM Troy, D Derossi, A Prochiantz… - Journal of …, 1996 - Soc Neuroscience
CM Troy, D Derossi, A Prochiantz, LA Greene, ML Shelanski
Journal of Neuroscience, 1996Soc Neuroscience
We previously showed that the downregulation of Cu/Zn superoxide dismutase (SOD1)
activity in PC12 cells by exposure to an appropriate antisense oligonucleotide causes their
apoptotic death. In this report, we used this model to examine the pathways by which SOD1
downregulation leads to death and to compare these pathways with those responsible for
death caused by withdrawal of trophic support. To improve delivery of the SOD1 antisense
oligonucleotide, we coupled it to a carrier “vector” peptide homologous to the third helix of …
We previously showed that the downregulation of Cu/Zn superoxide dismutase (SOD1) activity in PC12 cells by exposure to an appropriate antisense oligonucleotide causes their apoptotic death. In this report, we used this model to examine the pathways by which SOD1 downregulation leads to death and to compare these pathways with those responsible for death caused by withdrawal of trophic support. To improve delivery of the SOD1 antisense oligonucleotide, we coupled it to a carrier “vector” peptide homologous to the third helix of the Drosophila Antennapedia homeodomain. This caused not only efficient cellular uptake even in the presence of serum, but also inhibition of SOD1 activity and promotion of apoptosis at 100-fold lower concentrations of oligonucleotide. Death induced by SOD1 downregulation appeared to require the reaction of superoxide with nitric oxide (NO) to form peroxynitrite. In support of this, inhibitors of NO synthase, the enzyme responsible for NO synthesis, blocked death in our experiments, whereas NO generators and donors accelerated cell death. N-Acetylcysteine and chlorophenylthiol cAMP, which rescue PC12 cells and neurons from the withdrawal of nerve growth factor and other forms of trophic support, did not protect PC12 cells from SOD1 downregulation. In contrast, overexpression of bcl-2, which also rescues these cells form loss of trophic support, was equally effective in saving the cells in the SOD1 downregulation paradigm. Taken together with past findings, such observations suggest that SOD1 downregulation and withdrawal of trophic support trigger apoptosis via distinct initial mechanisms but may utilize a common final pathway to bring about death. Our findings may be relevant to the causes and potential amelioration of neuronal degenerative disorders caused by impaired regulation of cellular levels of NO and superoxide.
Soc Neuroscience