Migration of dendritic cells (DCs), antigen-presenting cells linking innate and adaptive immunity, is under powerful regulation by intracellular calcium([Ca²⁺]_i). Upon ligation of the chemokine receptors DCs respond with a fast increase of [Ca²⁺]_i, which involves Ca²⁺ release from the stores followed by Ca²⁺ entry through the store operated Ca²⁺ channels (SOCCs). The increase of [Ca²⁺]_i is rapidly terminated by Ca²⁺ extrusion, which is in DCs accomplished by via K⁺-independent (NCX) and K⁺-dependent (NCKX) Na⁺/Ca²⁺ exchangers. Both Ca²⁺ entry and Ca²⁺ extrusion are required for DC migration. Increase of [Ca²⁺]_i, activates the AMP-activated protein kinase (AMPK). The present study thus explored whether AMPK participates in the regulation of Ca²⁺ signalling in DCs. Experiments were performed in DCs isolated from AMPKa1-deficient (ampk^-/-) and wild type (ampk^+/+) mice. According to transwell chambers, following stimulation with CXCL12 (75ng/ml) migration was markedly enhanced in ampk^-/- as compared to ampk^+/+ DCs. According to Fura-2 fluorescence, CXCL12 (300 ng/ml)-induced increase of [Ca²⁺]_i was significantly higher in ampk^-/- than in ampk^+/+ DCs. When endosomal Ca²⁺ ATPase was inhibited with thapsigargin, SOCC activity was significantly increased in ampk^-/- DCs. Moreover, activity of both NCX and NCKX Na⁺/Ca²⁺ exchangers was significantly enhanced in ampk^-/- if compared to ampk^+/+ DCs. According to patch-clamp experiments, the NCX and NCKX currents were both significantly increased in ampk^-/- DCs. In conclusion, AMPK strongly suppresses DC migration, presumably through inhibition of both, SOCCs and Na⁺/Ca²⁺ exchangers in DCs.