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Members of the protein kinase D (PKD) family of serine/threonine kinases are known to exert diverse roles in neuronal stress responses. Here, we show the transient activation and nuclear translocation of endogenous PKD upon oxidative stress induced by H₂O₂ treatment in primary neuronal cultures. Using pharmacological inhibition, we show that PKD activity protects neurons from oxidative stress‐induced cell death. Although members of the canonical nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF kappaB) pathway were phosphorylated upon H₂O₂ treatment, it was found that the neuronal response to oxidative stress is not executed through the nuclear translocation and activity of RelA. On the other hand, we demonstrate for the first time in neuronal cells, the association of green fluorescent protein‐tagged kinase inactive PKD1 with mitochondrial membranes in vivo and the presence of …