1 We examined the mechanisms underlying leukotriene D 4 -(LTD 4 ) induced constriction of human small (300 ± 500 mm i.d.) bronchioles, and the e ect of LTD 4 on ion currents and Ca 2+ transients in smooth muscle cells (SMC) isolated from these bronchioles. 2 LTD 4 caused a concentration-dependent bronchoconstriction with an EC 50 =0.58+0.05 nM (n=7) which was not easily reversible upon washout. This bronchoconstriction was entirely dependent on extracellular Ca 2+ . 3 Blockade of L-type Ca 2+ channels with nifedipine (10 mM) reduced LTD 4 response by 39+2% (n=8), whilst La 3+ , Gd 3+ and SK&F 96,365 abolished LTD 4 -induced bronchoconstriction completely and reversibly, suggesting the majority of Ca 2+ entry was via non-selective cation channels. 4 Antagonists of PI-PLC (U73,122 and ET-18-OCH 3 ), PLD (propranolol) and PKC (cheleretrine and Ro31-8220) were without any e ect on LTD 4 -induced bronchoconstriction, whilst the PC-PLC inhibitor D609 caused complete relaxation. Inhibition of protein tyrosine kinase with tyrphostin A23 (100 mM) caused about 50% relaxation, although the inactive analogue tyrphostin A1 was without e ect. 5 In freshly isolated SMC from human small bronchioles LTD 4 caused a slow increase of intracellular Ca 2+ concentration, with a consequent rise of the activity of large conductance Ca 2+ -dependent K + channels and the amplitude of depolarization-induced outward whole-cell current. Again, no e ect of LTD 4 could be observed in the absence of extracellular Ca 2+ . 6 We conclude that LTD 4 causes constriction of these small bronchioles primarily by activating Ca 2+ entry via non-voltage gated channels, possibly by a PC-PLC mediated pathway.