Platelet-activating factor (PAF) is a very potent contractile agonist in guinea pig peripheral lung strips but is without effect on rabbit peripheral lung strips. Histological examination of guinea pig peripheral lung strips revealed a layer of smooth muscle cells in the visceral pleura that is not present in rabbit peripheral lung strips. Removal of the pleural surface of the guinea pig peripheral lung strips eliminated the ability of these tissues to contract to PAF, and the (removed) thin pleural strip contracted to PAF in a manner similar to that of intact strips. Removal of the pleural surface did not prevent these tissues from contracting to histamine or leukotriene D4 (LTD4), although potency of these agonists was somewhat reduced. The pleural smooth muscle cell layer is present throughout the pleural lining of the guinea pig lung but is thicker in lower than in upper lobes. PAF contractility is also reduced in upper compared with lower lung lobes. Thus we suggest that the pleural smooth muscle is the critical contractile element for PAF contraction of guinea pig peripheral lung strips, whereas other agonists such as histamine and LTD4 contract additional elements in these tissues.