The placenta supplies many nutrients to the fetus, including amino acids by active transport. Although the exact regulatory mechanism is unknown, a few studies have suggested a role for calcium in amino acid transport. Therefore, we examined the relationship between calcium and amino acid uptake by term human placental slices. Calcium depletion of slices significantly reduced uptake of α-aminoisobutyric (AIB), which is actively transported primarily by a sodium-dependent, carrier-mediated mechanism. Impairment of AIB uptake induced by calcium depletion was reversed by repletion with calcium but not with other divalent cations. In contrast, uptake of phenylalanine, which is transported primarily by a sodium-independent mechanism, was not affected by calcium depletion. Uptake of leucine and valine, which accumulate by both sodium-dependent and independent mechanisms, was partially affected by calcium depletion. Verapamil (10 μM), an inhibitor of transmembrane calcium flux, significantly reduced AIB uptake. Trifluoperazine, a calmodulin antagonist, also inhibited AIB uptake. Analysis of AIB uptake kinetic constants for control and calcium-depleted slices showed no change in the diffusion constant, a 37% reduction in Vmax, and a 2-fold increase in Km. The results indicate that calcium may be an important factor in the cellular regulation of active transport of amino acids.