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Protein kinase C expression and subcellular distribution in chronic myocardial ischemia: Comparison of two different canine models

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Abstract

We studied protein kinase C (PKC) isozyme expression and activity distribution in two models of chronically ischemic canine myocardium: (1) single vessel obstruction (SVO), produced by tight stenosis of LAD followed by preconditioning and acute ischemia (40 min); (2) three vessel obstruction (3VO), produced by LAD-stenosis and gradual occlusion of right coronary artery and left circumflex. In both models after 8 weeks of chronic ischemia the dogs were either sacrificed or had PTCA of the LAD with a follow up of another 4 weeks. Control dogs were sham operated. PKC activity was measured in subcellular fractions of tissue samples from anterior and posterior regions in the presence of histone and γ-[32P]-ATP. PKC isozymes were detected by Western blotting. All regions perfused by the obstructed coronaries were dysfunctional at 8 weeks when compared to baseline, with improvement of anterior wall function after PTCA of LAD. PKC activity was elevated in the membrane fraction of SVO, but unchanged in the 3VO model. PKCs α, ε, and ζ prevailed in cytosol fraction of the controls (cytosol/membrane ratios were ± 3.34, 1.38 and 4.56 for α, ε and ζ, respectively), consistent with PKC activity distribution, while δ was not detected. There was no significant difference between the groups concerning the relative membrane amount of the isozymes. PKCs α and ε were decreased in the cytosol fraction of both models at 8 weeks (for anterior region, by 56 and 57% in SVO and by 49 and 46% in 3VO, respectively) without there being any differences between anterior and posterior regions, and were low also in the PTCA group. PKC ζ distribution however varied between the two models. The amount of PKC ζ isozyme was downregulated by 45% after 8 weeks of chronic ischemia and returned towards the control values after PTCA in the anterior region of SVO, while it did not change in anterior wall after 8 weeks in 3VO but was significantly decreased (by 47%) in posterior region after PTCA. In conclusion, our results suggest modified PKC signalling in chronically ischemic canine myocardium.

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Authors and Affiliations

  1. Department of Cardiac Surgery, Katholieke Universiteit Leuven, Belgium

    Milena Matejovicova & Willem Flameng

  2. Department of Cardiology, Katholieke Universiteit Leuven, Belgium

    Bharati Shivalkar, Johan Vanhaecke & Monika Szilard

Authors
  1. Milena Matejovicova
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  2. Bharati Shivalkar
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  3. Johan Vanhaecke
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  4. Monika Szilard
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  5. Willem Flameng
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Matejovicova, M., Shivalkar, B., Vanhaecke, J. et al. Protein kinase C expression and subcellular distribution in chronic myocardial ischemia: Comparison of two different canine models. Mol Cell Biochem 201, 73–82 (1999). https://doi.org/10.1023/A:1007052232363

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