title: Myosin light chain kinase and Rho-kinase participate in P2Y receptor-mediated acceleration of permeability through the endothelial cell layer.
abstract: We have shown that P2Y receptor stimulation accelerates macromolecular permeation through the endothelial cell layer. To elucidate the mechanism of this acceleration, we examined the effects of ML-9, a myosin light chain kinase inhibitor, and Y-27632, a Rho-kinase inhibitor, on fluorescein isothiocyanate dextran (FD-4) permeation across the human umbilical vein endothelial cell monolayer. FD-4 permeation was analysed by high-performance liquid chromatography fluorescence detection. A P2Y receptor agonist, 2meS-ATP, enhanced the permeability of FD-4, which was inhibited by pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), a P2Y-receptor antagonist. The 2meS-ATP-induced increase in the permeability of FD-4 was significantly inhibited by ML-9. Also, Y-27632 prevented the 2meS-ATP-induced increase in the permeability of FD-4. Neither ML-9 nor Y-27632 influenced the spontaneous permeation of FD-4. These results suggest that phosphorylation of the myosin light chain may play an important role in the purinergic regulation of macromolecular permeation through the vascular endothelium.
authors.affiliation: Department of Pharmacology, School of Pharmaceutical Sciences, Mukogawa Women's University, Nishinomiya 663-8179, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Tanaka
authors.forename: Namie
authors.initials: N
authors.lastname: Nejime
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kubota
authors.forename: Satomi
authors.initials: S
authors.lastname: Kagota
authors.forename: Keiko
authors.initials: K
authors.lastname: Yudo
authors.forename: Kazuki
authors.initials: K
authors.lastname: Nakamura
authors.forename: Masaru
authors.initials: M
authors.lastname: Kunitomo
authors.forename: Koichi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Michio
authors.initials: M
authors.lastname: Hashimoto
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Shinozuka
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000577:Amides / Q000494:pharmacology; D001381:Azepines / Q000494:pharmacology; D002463:Cell Membrane Permeability; D002851:Chromatography, High Pressure Liquid; D003911:Dextrans / Q000493:pharmacokinetics; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D006801:Humans; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D013873:Thionucleotides / Q000494:pharmacology; D054460:rho-Associated Kinases
chemical_list: D000577:Amides; D001381:Azepines; D003911:Dextrans; D047908:Intracellular Signaling Peptides and Proteins; D011725:Pyridines; D018048:Receptors, Purinergic P2; D013873:Thionucleotides; C015219:fluorescein isothiocyanate dextran; C056218:ML 9; C108830:Y 27632; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D011732:Pyridoxal Phosphate; D000255:Adenosine Triphosphate; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D009219:Myosin-Light-Chain Kinase; D016650:Fluorescein-5-isothiocyanate; C008501:2-methylthio-ATP