title: alpha1-adrenoceptor stimulation potentiates L-type Ca2+ current through Ca2+/calmodulin-dependent PK II (CaMKII) activation in rat ventricular myocytes.
abstract: alpha1-Adrenoceptor stimulation (alpha1ARS) modulates cardiac muscle contraction under physiological conditions by means of changes in Ca2+ current through L-type channels (ICa,L) and Ca2+ sensitivity of the myofilaments. However, the cellular mechanisms of alpha1ARS are not fully clarified. In this study, we investigated the role of Ca2+/calmodulin-dependent PK II (CaMKII) in the regulation of ICa,L during alpha1ARS in isolated adult rat ventricular myocytes by using the perforated patch-clamp technique. CaMKII inhibition with 0.5 microM KN-93 abolished the potentiation in ICa,L observed during alpha1ARS by 10 microM phenylephrine. In the presence of PKC inhibitor (10 microM chelerythrine), the potentiation of ICa,L by phenylephrine also disappeared. In Western immunoblotting analysis, phenylephrine (> or =1 microM) increased the amount of autophosphorylated CaMKII (active CaMKII) significantly, and this increase was abolished by CaMKII inhibition or PKC inhibition. Also, we investigated changes in the subcellular localization of active CaMKII by using immunofluorescence microscopy and immunoelectron microscopy. Before alpha1ARS, active CaMKII was exclusively located just beneath the plasmalemma. However, after alpha1ARS, active CaMKII was localized close to transverse tubules, where most of L-type Ca2+ channels are located. From these results, we propose that CaMKII, which exists near transverse tubules, is activated and phosphorylated by alpha1ARS and that CaMKII activation directly potentiates ICa,L in rat ventricular myocytes.
authors.affiliation: Department of Physiology (II), Division of Cardiology, and Division of Molecular Cell Biology, The Jikei University School of Medicine, 3-25-8 Nishi-Shimbashi, Minato-ku, Tokyo 105-8461, Japan. o-uichi@jikei.ac.jp
authors.forename: Jin
authors.initials: J
authors.lastname: O-Uchi
authors.forename: Kimiaki
authors.initials: K
authors.lastname: Komukai
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Kusakari
authors.forename: Toru
authors.initials: T
authors.lastname: Obata
authors.forename: Kenichi
authors.initials: K
authors.lastname: Hongo
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sasaki
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kurihara
mesh_terms: D000818:Animals; D001596:Benzylamines / Q000494:pharmacology; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000737:chemistry*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006352:Heart Ventricles / Q000166:cytology*; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D032342:Muscle Cells / Q000166:cytology*; D018408:Patch-Clamp Techniques; D010656:Phenylephrine / Q000737:chemistry / Q000494:pharmacology; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology; D013997:Time Factors; D000096926:Benzenesulfonamides
chemical_list: D001596:Benzylamines; D002118:Calcium; D020746:Calcium Channels, L-Type; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D004791:Enzyme Inhibitors; D010656:Phenylephrine; D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; D018340:Receptors, Adrenergic, alpha-1; D013449:Sulfonamides; C072105:KN 93; D000096926:Benzenesulfonamides