title: Modulation of the c-Jun N-terminal kinase activity in the embryonic heart in response to anoxia-reoxygenation: involvement of the Ca2+ and mitoKATP channels.
abstract: Whether the response of the fetal heart to ischemia-reperfusion is associated with activation of the c-Jun N-terminal kinase (JNK) pathway is not known. In contrast, involvement of the sarcolemmal L-type Ca2+ channel (LCC) and the mitochondrial KATP (mitoKATP) channel has been established. This work aimed at investigating the profile of JNK activity during anoxia-reoxygenation and its modulation by LCC and mitoK(ATP) channel. Hearts isolated from 4-day-old chick embryos were submitted to anoxia (30 min) and reoxygenation (60 min). Using the kinase assay method, the profile of JNK activity in the ventricle was determined every 10 min throughout anoxia-reoxygenation. Effects on JNK activity of the LCC blocker verapamil (10 nM), the mitoK(ATP) channel opener diazoxide (50 microM) and the blocker 5-hydroxydecanoate (5-HD, 500 microM), the mitochondrial Ca2+ uniporter (MCU) inhibitor Ru360 (10 microM), and the antioxidant N-(2-mercaptopropionyl) glycine (MPG, 1 mM) were determined. In untreated hearts, JNK activity was increased by 40% during anoxia and peaked fivefold relative to basal level after 30-40 min reoxygenation. This peak value was reduced by half by diazoxide and was tripled by 5-HD. Furthermore, the 5-HD-mediated stimulation of JNK activity during reoxygenation was abolished by diazoxide, verapamil or Ru360. MPG had no effect on JNK activity, whatever the conditions. None of the tested pharmacological agents altered JNK activity under basal normoxic conditions. Thus, in the embryonic heart, JNK activity exhibits a characteristic pattern during anoxia and reoxygenation and the respective open-state of LCC, MCU and mitoKATP channel can be a major determinant of JNK activity in a ROS-independent manner.
authors.affiliation: Department of Physiology, Faculty of Biology and Medicine, University of Lausanne, 7 rue du Bugnon, 1005 Lausanne, Switzerland. alexandre.sarre@medecine.unige.ch
authors.forename: Alexandre
authors.initials: A
authors.lastname: Sarre
authors.forename: Stéphany
authors.initials: S
authors.lastname: Gardier
authors.forename: Fabienne
authors.initials: F
authors.lastname: Maurer
authors.forename: Christophe
authors.initials: C
authors.lastname: Bonny
authors.forename: Eric
authors.initials: E
authors.lastname: Raddatz
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D002645:Chickens; D004795:Enzyme Stability; D006321:Heart / Q000196:embryology*; D006352:Heart Ventricles / Q000196:embryology / Q000201:enzymology; D000860:Hypoxia / Q000201:enzymology*; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D008954:Models, Biological; D015221:Potassium Channels / Q000378:metabolism*; D012508:Sarcolemma / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D015221:Potassium Channels; C117780:mitochondrial K(ATP) channel; C034703:mitochondrial calcium uniporter; D048031:JNK Mitogen-Activated Protein Kinases; D002118:Calcium