Preeclampsia is thought to arise from inadequate cytotrophoblast migration and invasion of the maternal spiral arteries, resulting in placental ischemia and hypertension. Evidence suggests that altered expression of Epithelial Na⁺ Channel (ENaC) proteins may be a contributing mechanism for impaired cytotrophoblast migration. ENaC activity is required for normal cytotrophoblast migration. Moreover, βENaC, the most robustly expressed placental ENaC message, is reduced in placentas from preeclamptic women. We recently demonstrated that heme oxygenase-1 (HO-1) protects against hypertension in a rat model of placental ischemia; however, whether HO-1 regulation of βENaC contributes to the beneficial effects of HO-1 is unknown. The purpose of this study was to determine whether βENaC mediates cytotrophoblast migration and if HO-1 enhances ENaC-mediated migration. We showed that placental ischemia, induced by reducing uterine perfusion, suppressed, and HO-1 induction restored βENaC expression in ischemic placentas. Using an in vitro model, we found that HO-1 induction, using cobalt protoporphyrin, stimulates cytotrophoblast βENaC expression by 1.5 and 1.8-fold (10 and 50 µM). We then showed that silencing of βENaC in cultured cytotrophoblasts (BeWo cells), by expression of dominant-negative constructs, reduced migration to 56 ± 13% (p<0.05) of control. Importantly, HO-1 induction enhanced migration (43 ± 5% of control, p<0.05), but the enhanced migratory response was entirely blocked by ENaC inhibition with amiloride (10 µM). Taken together, our results suggest that βENaC mediates cytotrophoblast migration and increasing βENaC expression by HO-1 induction enhances migration. HO-1 regulation of cytotrophoblast βENaC expression and migration may be a potential therapeutic target in preeclamptic patients.