We recently demonstrated that superoxide (O₂^•−) is a key signaling intermediate in central angiotensin II (Ang II)-elicited blood pressure and drinking responses, and that hypertension caused by systemic Ang II infusion involves oxidative stress in cardiovascular nuclei of the brain. Intracellular Ca²⁺ is known to play an important role in Ang II signaling in neurons, and it is also linked to reactive oxygen species mechanisms in neurons and other cell types. However, the potential cross-talk between Ang II, O₂^•−, and Ca²⁺ in neural cells remains unknown. Using mouse neuroblastoma Neuro-2A cells, we tested the hypothesis that O₂^•− radicals are involved in the Ang II–induced increase in intracellular Ca²⁺ concentration ([Ca²⁺]_i) in neurons. Ang II caused a rapid time-dependent increase in [Ca²⁺]_i that was abolished in cells bathed in Ca²⁺-free medium or by pretreatment with the nonspecific voltage-gated Ca²⁺ channel blocker CdCl₂, suggesting that voltage-sensitive Ca²⁺ channels are the primary source of Ang II–induced increases in [Ca²⁺]_i in this cell type. Overexpression of cytoplasm-targeted O₂^•− dismutase via an adenoviral vector (AdCuZnSOD) efficiently scavenged Ang II–induced increases in intracellular O₂^•− and markedly attenuated the increase in [Ca²⁺]_i caused by this peptide. Furthermore, adenoviral-mediated expression of a dominant-negative isoform of Rac1 (AdN17Rac1), a critical component for NADPH oxidase activation and O₂^•− production, significantly inhibited the increase in [Ca²⁺]_i after Ang II stimulation. These data provide the first evidence that O₂^•− is involved in the Ang II–stimulated influx of extracellular Ca²⁺ in neural cells and suggest a potential intracellular signaling mechanism involved in Ang II–mediated oxidant regulation of central neural control of blood pressure.