The L-type Ca²⁺ channel expressed in gastrointestinal smooth muscle is mechanosensitive. Direct membrane stretch and shear stress results in increased Ca²⁺ entry into the cell. The mechanism for mechanosensitivity is not known, and mechanosensitivity is not dependent on an intact cytoskeleton. The aim of this study was to determine whether L-type Ca²⁺ channel mechanosensitivity is dependent on tension in the lipid bilayer in human jejunal circular muscle myocytes. Whole cell currents were recorded in the amphotericin perforated-patch configuration and lysophosphatidyl choline (LPC), lysophosphatidic acid (LPA) and choline used to alter differentially the tension in the lipid bilayer. Shear stress (perfusion at 10 ml/min) was used to mechanostimulate L-type Ca²⁺ channels. In the presence of LPC (large head-to-tail proportions) but not LPA or choline, the increase in L-type Ca²⁺ current induced by shear stress was greater than that observed in the control perfusion. The increased peak Ca²⁺ current also did not return to baseline levels as in control conditions. Furthermore, steady-state inactivation kinetics were altered in the presence of LPC, leading to a change in window current. These findings suggest that changes in tension in the plasmalemmal membrane can be transmitted to the mechanosensitive L-type Ca²⁺ channel leading to altered activity and Ca²⁺ entry in the human jejunal circular muscle myocyte.