We investigated the role of src family kinases (srcFK) in agonist-mediated Ca²⁺-sensitization in pulmonary artery and whether this involves interaction with the rho/rho-kinase pathway.

Intra-pulmonary arteries (IPAs) and cultured pulmonary artery smooth muscle cells (PASMC) were obtained from rat. Expression of srcFK was determined at the mRNA and protein levels. Ca²⁺-sensitization was induced by prostaglandin F_2α (PGF_2α) in α-toxin-permeabilized IPAs. Phosphorylation of the regulatory subunit of myosin phosphatase (MYPT-1) and of myosin light-chain-20 (MLC₂₀) and translocation of rho-kinase in response to PGF_2α were also determined. Nine srcFK were expressed at the mRNA level, including src, fyn, and yes, and PGF_2α enhanced phosphorylation of three srcFK proteins at tyr-416. In α-toxin-permeabilized IPAs, PGF_2α enhanced the Ca²⁺-induced contraction (pCa 6.9) approximately three-fold. This enhancement was inhibited by the srcFK blockers SU6656 and PP2 and by the rho-kinase inhibitor Y27632. Y27632, but not SU6656 or PP2, also inhibited the underlying pCa 6.9 contraction. PGF_2α enhanced phosphorylation of MYPT-1 at thr-697 and thr-855 and of MLC₂₀ at ser-19. This enhancement, but not the underlying basal phosphorylation, was inhibited by SU6656. Y27632 suppressed both basal and PGF_2α-mediated phosphorylation. The effects of SU6656 and Y27632, on both contraction and MYPT-1 and MLC₂₀ phosphorylation, were not additive. PGF_2α triggered translocation of rho-kinase in PASMC, and this was inhibited by SU6656.

srcFK are activated by PGF_2α in the rat pulmonary artery and may contribute to Ca²⁺-sensitization and contraction via rho-kinase translocation and phosphorylation of MYPT-1.