Up-regulation and redistribution of protein kinase C-δ in chronically hypoxic heart.
The adaptation to chronic hypoxia confers long-lasting cardiac protection against acute ischemia-reperfusion injury. Protein kinase C (PKC) appears to play a role in the cardioprotective mechanism but the involvement of individual PKC isoforms remains unclear. The aim of this study was to examine the effects of chronic intermittent hypoxia (CIH; 7,000 m, 8 h/day) and acute administration of PKC-δ inhibitor (rottlerin, 0.3 mg/kg) on the expression and subcellular distribution of PKC-δ and PKC-ε in the left ventricular myocardium of adult male Wistar rats by Western blot and quantitative immunofluorescence microscopy. CIH decreased the total level of PKC-ε in homogenate without affecting the level of phosphorylated PKC-ε (Ser729). In contrast, CIH up-regulated the total level of PKC-δ as well as the level of phosphorylated PKC-δ (Ser643) in homogenate. Rottlerin partially reversed the hypoxia-induced increase in PKC-δ in the mitochondrial fraction. Immunofluorescent staining of ventricular cryo-sections revealed increased co-localization of PKC-δ with mitochondrial and sarcolemmal membranes in CIH hearts that was suppressed by rottlerin. The formation of nitrotyrosine as a marker of oxidative stress was enhanced in CIH myocardium, particularly in mitochondria. The expression of total oxidative phosphorylation complexes was slightly decreased by CIH mainly due to complex II decline. In conclusion, up-regulated PKC-δ in CIH hearts is mainly localized to mitochondrial and sarcolemmal membranes. The inhibitory effects of rottlerin on PKC-δ subcellular redistribution and cardioprotection (as shown previously) support the view that this isoform plays a role in the mechanism of CIH-induced ischemic tolerance.
Hlaváčková
Markéta
M
Department of Cell Biology, Faculty of Science, Charles University in Prague, Vinicna 7, 128 00, Prague 2, Czech Republic. hlavacko@natur.cuni.cz
Kožichová
Kristýna
K
Neckář
Jan
J
Kolář
František
F
Musters
René J P
RJ
Novák
František
F
Nováková
Olga
O
D000818:Animals; D002908:Chronic Disease; D000860:Hypoxia / Q000201:enzymology*; D008928:Mitochondria / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation; D020011:Protective Agents; D051745:Protein Kinase C-delta / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012508:Sarcolemma / Q000378:metabolism; D014018:Tissue Distribution; D015854:Up-Regulation
D020011:Protective Agents; D047428:Protein Kinase Inhibitors; D051745:Protein Kinase C-delta