Species differences in the effects of prostanoids on MAP kinase phosphorylation, myosin light chain phosphorylation and contraction in bovine and cat iris sphincter smooth muscle

There is evidence from our own laboratory and that of others that EP-receptor ligands are strong contractile agonists in bovine iris sphincter and that FP-receptor agonists are strong contractile agonists in cat iris sphincter. Here, we have investigated the effects of prostaglandin (PG) receptor agonists of the FP-, EP-, TP- and DP-class on myosin light chain (MLC) phosphorylation, p42/p44 MAP kinase phosphorylation and contraction in the iris sphincter of bovine and cat. Using three signal transduction mechanism assays, namely MLC phosphorylation, MAP kinase phosphorylation and contraction, we demonstrated that in bovine iris sphincter the rank order of potency of the PG agonists in the contractile and MLC phosphorylation assays is as follows: E 2>U46619> F 2 α >D 2, and in cat F 2 α > D 2> E 2>U46619. In the MAP kinase assay, in bovine iris sphincter the rank order of potency is E 2> F 2 α and in cat F 2 α > E 2. These conclusions are supported by the following findings: (1) In the contractile assay, in the bovine sphincter the EC 50s for PGF 2 α , PGE 2, U46619 and PGD 2 were found to be 1.4×10 –7, 5.0×10 –9, 9.0×10 –9 and 1.3×10 –6 M, respectively, and the corresponding values in the cat were 1.9×10 –8, 2.3×10 –7, 1.5×10 –6 and 6.9×10 –8 M, respectively. (2) In the MLC phophorylation assay, in the bovine sphincter PGF 2 α , PGE 2, U46619 and PGD 2 increased MLC phophorylation by 118%, 165%, 153% and 72%, respectively, and the corresponding values in cat were 175%, 99%, 90% and 95%, respectively. (3) In the MAP kinase assay, in the bovine iris sphincter PGF 2 α and PGE 2, increased MAP kinase phosphorylation by 276% and 328%, respectively, and the corresponding values in cat were 308% and 245%, respectively. The data presented demonstrate pronounced species differences in the effects of the prostanoids on the MLC kinase signaling pathway in bovine and cat irides and furthermore confirm the existence of FP-receptors in that of the bovine.