Mode of action of thrombin in the rabbit aorta.

1. Thrombin is a vasoactive protease that elicits the contraction of the rabbit aorta by activating a G-protein coupled receptor through cleavage of its N-terminal extracellular domain. Synthetic peptides corresponding to the newly exposed N-terminus, following thrombin cleavage, have been shown to reproduce some of the activities of thrombin in the rabbit aorta. 2. Intracellular pathways involved in the contractile response of the rabbit aorta to thrombin and synthetic peptides were examined by use of a series of inhibitors. A similar method was applied to characterize the mitogenic effect of thrombin on cultured smooth muscle cells (SMCs) derived from the same tissue. 3. Results from this study indicate that the contractile response of the rabbit aorta to thrombin is dependent on the activation of protein kinase C (PKC) and independent of extracellular calcium. The contractile response to thrombin can be fully reproduced by peptide agonists related to the N-terminal receptor sequence. However, subtle differences seem to exist between the mechanism of the contractile effect of thrombin and of the synthetic peptides, as both PKC activation and extracellular calcium were found to participate in the contractile effect of the synthetic peptides. 4. In cultured SMCs, both thrombin and the synthetic peptides increased inositol phosphate turnover; however, only thrombin elicited a mitogenic effect, which occurs at thrombin concentrations well below those needed to increase inositol phosphate turnover significantly. Activation of a tyrosine kinase pathway is involved in the mitogenic effect of thrombin on aortic SMCs. 5. Altogether these results suggest the existence of subtle differences between the mode of action of thrombin and of synthetic peptides related to the N-terminal thrombin receptor sequence, in the rabbit aorta.